Bioanalytical Trends: Client Needs, Platforms & Regulatory Requirements

The main discussion in Episode #101 of “The Weekly Bioanalysis” focuses on how bioanalytical testing continues to evolve alongside increasingly complex drug modalities, with our co-hosts, Dom and John, exploring the technologies, methodologies, and regulatory trends shaping modern bioanalytical laboratories. They discuss how advances in mass spectrometry, including the transition to newer SCIEX 6500 and 7500 platforms, along with emerging technologies such as the Alamar NULISA system, are enabling dramatically greater sensitivity while requiring smaller sample volumes and supporting more challenging biomarker studies. The conversation also examines the growing importance of assay selectivity, fit-for-purpose validation strategies, and the industry's ongoing debate surrounding single replicate analysis versus traditional duplicate testing as laboratories seek to balance efficiency, cost, and scientific rigor. Throughout the discussion, the hosts emphasize that no single analytical platform solves every challenge, making it increasingly important for CROs to maintain a diverse suite of technologies capable of addressing a wide range of client needs.

“The Weekly Bioanalysis” is a podcast dedicated to discussing bioanalytical news, tools and services related to the pharmaceutical, biopharmaceutical and biomarker industries. Every month, KCAS Bio will bring you another 60 minutes (or so) of friendly banter between our two finest Senior Scientific Advisors as they chat over coffee and discuss what they’ve learned about the bioanalytical world the past couple of weeks. “The Weekly Bioanalysis” is brought to you by KCAS Bio.

KCAS Bio is a progressive growing contract research organization of well over 250 talented and dedicated individuals with growing operations in Kansas City, Doylestown, PA, and Lyon, France, where we are committed to serving our clients and improving health worldwide. Our experienced scientists provide stand-alone bioanalytical services to the pharmaceutical, biopharmaceutical, animal health and medical device industries.

Transcript

SPEAKER_01 0:03

Welcome to the weekly bioanalysis, a KCast Bio podcast.

SPEAKER_00 0:10

Hello and welcome to the 101st episode of the Weekly Bioanalysis, the official podcast of KCast Bio. KCAS Bio is a bioanalytical CRO serving the pharmaceutical and biopharmaceutical industries for over 45 years. My name is Dominic Quarino, and I've been in the LBA and biomarker space for around 30 years. I'm here with my co-host, John Perkins.

SPEAKER_02 0:30

Hi, I'm John Perkins, and I've been in the bioanalytical LCMS space for around 30 years, too.

SPEAKER_00 0:36

John and I are members of our growing scientific services team here at KCast Bio. Either or both of us are available to answer any questions you may have about our services or this podcast or any of any other topics of interest that you may have. We are thrilled to have you listening to the 101st episode of the podcast, which is now available virtually everywhere. Wherever you choose to find and play your podcast, you can now likely find the weekly bioanalysis. We welcome all of you, whether you are joining us for the first time or if you're a regular listener. Today's podcast will be a review of our latest news and resources, and then a focus on a topic of our choosing before discussing any feedback we've had from you. We're constantly looking for new topics and we'd be happy to discuss something that you want us to cover. So again, we're fired up to have you here, and we're looking forward to a fun episode. To kick us off for podcast 101, John is going to go over today's podcast topics a bit before we jump into the news and resources. John?

SPEAKER_02 1:37

Yes, today we thought we'd look at trends in bioanalysis. Evolution of client needs instrumentation platforms and regulatory requirements. So it's it's what we're doing to adapt to the the changing state of bioanalysis and and the problems our clients thend our way.

SPEAKER_00 1:56

Yeah, great topic, John. We're exciting podcast to go over, but why don't you get us uh started with the news and notes first?

SPEAKER_02 2:03

First of all, you never guess what. We're back to the GLP ones. Um this one, GLP ones may combat depression through a mood busting gut microbe. Um so obviously we talk about GLP ones a lot simply because they're uh dominating the market in terms of best-selling drugs, etc. And there's a lot of discussion about different indications. Um there's been a lot of question marks about them. In some they seem to have antidepress some people seem to have antidepressant effects, while others they've sort of thought they might um be linked to worse mental health, but it's not not clear now in actual fact, as the article states, um, the the sort of negative mental health was on labels and has been taken or has been asked to be removed. Um but this is a team of researchers in China who f who basically found that mice treated with GLP1s, laraglitide and somaglitide, saw um lactobactobacillus delbrucci bacteria flourish in their guts with the microbes churning at a compound that alleviates symptoms of depression. So it it's sort of showing a mechanism whereby they they they might actually help depression. And um it's like it's although it's like you think this should be the brain, it this is it's all about gut health and um they they so in in some of the precursor they looked at blood from patients with major depressive disorder and they found that natural levels of GLP1 were lower in with those in depression. When they explored further by injecting laraglitide into depressed mice, they found that the peptide built up in the intestine rather than the brain, and that's what pushed them to say, well, what's going on? Looked at um the actually mice feces and found higher levels of this lactobacillus delbrucci, and um it basically that that bacterium uh that yeah that that bacterium basically supports the synthesis of cannabis-like molecule called 2 arachidonoglycerol or 2AG, which has a soothing effect on the brain. I'll pass it back to you because I've I've had I think I'm done with the pronunciations for a while.

SPEAKER_00 4:23

Yeah, no, uh um, you know, it's become uh gut gut health seems to be all the rage, John, right? It's uh sure it's something where um I think it the the sheer it makes a lot of sense on the surface, but no one thinks about hey, is are my is my gut healthy? And if I'm not healthy, the one of the last things people are gonna think about is like their stomach, although I think now it's becoming much more aware of just how important your gut health is. I mean, it's like its own immune system, it's a very large portion of your body, and so it kind of makes sense that if you're got some bacteria going on somewhere, it could be causing depression or pain or whatever it might be. Um, and it's but it's a fascinating story. These GLP1s continue to be like magic drugs, right? Like not only are they it will lose weight, they're they're now like potentially because John, if I'm reading it right, they're they're actually they removed a warning label, right? I don't know if you want to talk about that.

SPEAKER_02 5:20

Yeah, yeah, it's been requested to to remove the warning label. I think it was the FDA drove that. And then actually both um Lily is definitely looking at um one of their GLP, it's actually a dual GLP1 GIP, um Brenn Brenipatide, um uh administered with antidepressant therapy in terms of helping adults with major depressive disorder, and um novonordisc are similarly looking at um smaglide for major depressive disorders. So it is something that that you know um the the the lead manufacturers are looking at how can these how can these um molecules help? So it's it's yeah, it I just I thought it's fascinating. We've talked about this and yet again it's another this may be another indication of where these these these drugs can help. I mean I think it's interesting that they're talking about this and saying the pathway has nothing to do with the GLP1 receptor, which may explain why different studies have come to a different conclusion about the effect GLP1 drugs have on mental health. Um GLP1 analogues can activate central GLP1 receptors in the brain regions involved in stress and anxiety, and so the net effect on mood may depend on the balance between the imposing mechanisms of the what's attach what's affecting the GLP one receptor versus your gut health piece and it's it it like I say it's it's fascinating and I I obviously I watch watch these drugs with interest because yeah, just seeing the different varieties of effects that we weren't thinking of in the first place. And and we I mean no one was thinking obesity when these were first developed. They were it really it was you know what can we do for diabetes and this this is all downstream. Oh, we found this and we found this and found this, and so it's it's like an evolving um understanding of what these drugs may possibly contribute to us, and and yeah, that the fascinating.

SPEAKER_00 7:17

Well, I mean it's it's uh the label, so these GLP1 labels had a um potential warning around depression and suicide, right? Now it's become like 180, right? Like now these they're actually saying that this um brintuberum tubamad, right? The dual GLP1 GIP from Lily, that it co-administered with antidepressant may help c prevent depression, right? So it's like come full circle, and it is it's just fascinating to think about how um warning labels change. Um, as we're beginning to learn more about GLP1s, they've cured they're helping people with late weight loss. Now there's this potential benefit for you know um uh antidepression. I I feel like it's too still to be determined just how cheap people these drugs are. I mean, I I think there's some things that talk about maybe helping with remedy um um uh migraines and things like that. So I it's just it just it it makes sense now when you think about how hard it is to tell how effective and what it's from if someone's gut health is different. But yeah, gut health is playing a factor here, and I think in all of us, I think that we can universally agree to. It's just if the the fact that gut health that's a hard thing to study, right? Understanding everybody's gut is going to be different. And then think about the the research behind this, John, where they kind of took this lactobacillus uh kind of bacteria, and now they're starting to really it's really great basic research, right, John. That that's some basic research in mice, and they're they're finding out that, oh wow, this this abundant, this excess bacteria might be something that is causing some sort of depression, and lo and behold, the GLP ones knock it down, right? And so it it it's unbelievable how impactful they are. And I just maybe just in closing this out, it's still like one of all this work's done in male mice, right, John?

SPEAKER_02 9:13

I think that's yes, and so they don't know in female, yeah. It is worth emphasizing this is just in mice, so we're it's still in the early stages of of seeing if this even has potential for people, but again, it's identifying a potential mechanism where it might have applicability to people in the long term.

SPEAKER_00 9:31

You know, it's yet another story about GLP1, John. I got a feeling it won't be the last, right?

SPEAKER_02 9:36

I mean, I think we're gonna constantly and and and I wasn't really really focusing too much on the label change, but I like I say that request did come from the FDA because they had it. There was an analysis at the agency, found no connection between the medicines and suit suicidality. So that that's why there was the the change there. Um yeah, I'm I'm not here, I'm not I certainly not expounding GLP ones as as miracle medicines, but it is fascinating where they might be able to contribute and um where it help generally help people.

SPEAKER_00 10:07

I think that story is just beginning, not ending, right, Josh? Exactly. Speaking of stories, tell us about the next uh let's let's move on.

SPEAKER_02 10:14

This is actually um it's again very interesting. This is this is I suppose this is addressing a much older issue. Um, this is actually new antibodies stop measles from stabbing cells, um, study fines. And although we've had a measles vaccine for 60 years, much still remains unknown about how exactly the shot generates protection against the virus. Um, scientists actually looked at the immune system of vaccinated people um to identify antibodies. Um it was looking at the blood of 20 people previously given the MMR vaccine, and they they identified four antibodies that cling to the measles virus surface and prevent it from infecting the cells. So the the the potential so um she said that that was real she said that was all real sorry, the sapphire uh Erica Ullman sapphire um is a structural biologist at La Jolla Institute for Immunology who led the study and she basically said finding these antibodies was easy because um yeah measles doesn't have a as thick a protective layer of sugar molecules around it as other viruses do, so it was I think easier to pick these apart. Um but the the although we've had this vaccine for a long time, there are actually populations such as the immunocompromised and infants who can't take it. So actually having these antibodies available as a as a prophylactic may actually give another mechanism to help people where there is if if uh a measles infection takes place. So I it's it's sort of it's it's interesting because the article states they never really looked into the mechanism because they came up with the um because came up with the vaccine a long time ago where there wasn't the ability to look into the mechanism. We where our tools have advanced so much more. Now we can turn around and look and say, well, what's going on? How does this vaccine work? And and from looking at how does this vaccine work, possibly identifying other approaches that that would then be um supplemental to the vaccine to actually widen the applicability or the yeah, the application of of um you know treating measles uh if we if there's an outburst outbreak.

SPEAKER_00 12:36

Yeah. Well, I mean, let's just take a make sure everybody understands that vaccine debuted in 1963, right?

SPEAKER_02 12:44

So it wasn't a lot of uh good year 1963, because I debuted in 1960.

SPEAKER_00 12:51

Well, uh, but the fact that it was at the it's amazing how highly effective this was, and it isn't until 2026 that someone you know has uh the sapphire and Dr. Sapphire there has has been able to identify the four antibodies, right? Through a retrospective study, they've kind of found four antibodies that bind to the uh MMR uh kind of proteins, right? But it's in what they underlie in the story is because it worked in '63, no one's really questioned it, right, John, until they kind of came along and helped identify them. But the real breakthrough here is that now they think they can have prophylactic antibodies for preventing MMR. And that's important because there's, you know, Canada, the the article references how Canada's lost their status of eliminating it. So there's like more than 10 million cases of measles being outbreaked, even though I think people are getting vaccines for it. There's still kind of these breakthrough cases where now this these antibodies become potential therapeutics, right? So I think that's really and then will it ever replace the vaccine?

SPEAKER_02 13:59

I don't think they're arguing that, although I think it really is there to help other populations.

SPEAKER_00 14:05

Yeah.

SPEAKER_02 14:06

And again, it's worth pointing out this is this is this is in done in cotton rats, which I've actually not heard of before. But again, it's similar to the previous thing where it's it's been done in rodents and showing some applicability. Um, because uh let me see, yeah. So the antibodies were able to halt virus replication in cotton rats when given either before or as long as two days after exposure to measles. And so the four antibodies actually have different mechanisms. To the antibodies target measles, homoglutinin, um, surface protein, which the virus uses to attach to cells, while the others bind to the fusion protein used to physically pierce the cell to grant the virus entry. The fusion protein springs forward in an irreversible way, like a spearfishing rod to change shape and drive its itself into a human cell. So that's where the headline comes from about um, you know, stopping measles from from from stabbing cells, so it's it's it's in interrupting that fusion protein.

SPEAKER_00 15:10

Yeah, well, I mean, no, I don't think anybody's really thinking about MMR research or vaccines, but here we are with 10 million people apparently having it. So holy smokes, that's a lot, right?

SPEAKER_02 15:21

And so actually the the 10 million is in Canada, it's worldwide there are 10 million cases of measles every year.

SPEAKER_00 15:29

Yeah, yeah, which is still high though, given a vaccine. Absolutely, yeah. Yeah, and so I'm guessing somewhere in there they thought, hey, we may, you know, that that probably was the you know the the activation energy to get people to look at, hey, what are these things doing? And you're absolutely right, there's still caution to be made, but this is this is hopeful for um the next wave of let's say prophylactic therapeutics, not necessarily vaccines, but it's it it would be nice if of you know a physician who has someone who's been vaccinated and they end up getting measles or uh you know, or even if I say they don't have it and they get it, uh there's not a lot of therapies for it. This could be really groundbreaking for it, right? But I I do think you're right in that this cotton mouse model is not a not a disease model, so they kind of call it an infection model. But what I'm driving at is they say they reference non-human primates in ferrets. And I don't know if you knew that, John, but ferrets, yep, ferrets are great models for um any sort of vaccines. A lot of your AAV models early on were being done in in ferrets and still are, but it but just for whatever reason the ferret immune system is very predictive of of human immune systems for vaccines where this cotton rat, and never heard of it myself, John, so I I don't know anything about that cotton rat, but it seems pretty, pretty um, you know, kind of maybe not the best model as they indicate, but it's more to come. And but like it's it's a really who who it goes back to what we always like to talk about and how that those basic research labs are really critical to advancing diseases. And more importantly, if something does kind of come up, there's some lab somewhere with something as a potential therapy, right? And this is a classic example of of somebody being ready for well, I think there is somewhat of an immediate need, but you know, this this has incredible benefit if if they can indeed start to identify anybody's that could can treat MMR diseases. Yeah, yeah.

SPEAKER_02 17:30

Yeah. So sorry, John. Let's move on. Yeah. So it's two two down. Um this one now another another um another topic we we we visit regularly. This is protective genes sent to the liver created. Sorry, let me rephrase that. Protective genes sent to the liver cleared brain amyloid in in mice. And so uh this this is moving into Alzheimer's where we think about um you know the use the the presence of misfolded amyloid protein in the brain, um, but most of our approaches uh tend to be to uh penetrate the blood brain barrier to address, you know, obviously what's going on in the brain. But this study suggests that crossing the blood brain barrier might not be necessary at all to clear amyloid plaques, and that a one-time gene therapy could one day prove an effective treatment for the for for Alzheimer's. This was done at Army Medical University in Chongq, Chongqing, um, China. Um they they had a a gene for protective protein was successfully delivered to mice, leading to reduced plaques and cognitive improvements. Unlike other gene therapy approaches, this technique uses a standard liver-targeted vector rather than trying to get into the brain. Um it's basically they know that uh a a variant of the apolipoprotein e-gene known as ApoE4 is associated with a higher risk of developing Alzheimer's disease. But more recently, protective versions of the gene have been identified, most notably a variant called ApoE3 Christchurch. And this was the the the the potential of the Christchurch variant was was uh sort of identified when there was a woman who had copies of a mutated presenylum gene. Mutations in the gene are known to cause early onset uh early onset Alzheimer's that typically leaves to cognitive impairment beginning around age 44. But because she had two copies of the um ApoE3 Christ chart, she didn't belong begin developing symptoms until her 70s. So um they the researchers basically package the gene into a viral vector, inject it into male mice with the risky um ApoE4 variant, and um so they they say ApoE is also expressed in liver, and that's where the gene therapy went to. Treated mice showed smaller amyloid plaques, less brain inflammation, and perform better in maize memory tests. I will pass it to you because I'm I'm somewhat struggling to digest this.

SPEAKER_00 20:14

Yeah, it it's uh it's a it's so John. I I think the gist of it is that you can you can treat Alzheimer's, which as we all know is a neurodegenerative disorder, and we got the blood-brain barrier, which is a real problem, but you can treat it through activating therapeutics in the liver, right? That's what this is shown. And then the liver somehow, whenever it's producing these protective proteins, it's gonna make its way into the brain and and reduce the the plaques, right? That's the whole the Apo, I believe the ApoE3 helps do that. Um, but it's it's really um it's it's it's kind of it's get it gets back to just how um creative and I don't I don't know if luck's involved, but this basic researchers are doing these things over here that normally we might of some of us that have been focused primarily on like drug development and you know late-stage can candidates and preclinical studies and clinical studies, this is way upstream of that. Sure. That true basic research. And you know, the obviously these investigators were incredibly surprised that you know therap therapies, therapeutic area uh things that were augmenting the liver were cure in Alzheimer's.

SPEAKER_02 21:31

So it's if nothing at all, the these types of articles, because it is it's hard to untwine exactly how this is working because the yeah, and there's an independent, you know, uh they personally asked for comment, a guy called Jason Ulrich, uh neurologist at Washington University in St. Louis, who's sort of saying, I need to know more about this mechanism as to is has this got potential or not. According to the Chinese researchers, their thought is, and it's all theoretical. reticle at this point. Um the ApoE3 Christchurch clears amyloid out of the blood. This gene variant helps deliver an immune cells called monocytes, remove amyloid from the blood, and because the brain and blood maintain an amyloid balance, this leads to an exodus of the disease causing protein from the brain. That's that's the theory behind it. So obviously their their next step is go to non-human primates, see if they're seeing the same effect there, which comes back to the previous article where we're saying that you know obviously mice aren't a great model for people but non-human primates are much more relevant for us. And um yeah so what it's a it's a watch this space kind of thing but it's sort of fascinating that you know you've got potentially going for the liver could have I mean I you look it's all about the interchangeability within your body but it's still I mean it's still fascinating that it could be a mechanism because we talk a lot about you know trying to get across the blood brain barrier with large molecules and all that kind of stuff and and at least for this particular application this this alternative route may may have potential. Who knows?

SPEAKER_00 23:04

We'll we'll we'll see yeah I I think there's been 15 20 years of research dedicated to uh neurodegenerative uh type of medicines and there's been very few successes uh there are some approvals but I I don't you know they're not the most effective at preventing or or treating it and you know it's always been uh difficult because of the blood brain barrier because of the just the different I think the different uh every human's like a snowflake John right so it doesn't matter how you know the the disease itself doesn't have any etiology that's like a single point that's why there's sure been over the years biomarkers like A beta proteins tau and phosphotau and even APOs. So they got all these biomarkers that it and even when they think they've discovered one given biomarker that if it's elevated that's going to be indicative but what they're finding out is no you can have people at normal levels of that but there's always these like redundancies and some sort of ratio of I really do think it's like a cocktail of biomarkers that kind of creates the disease itself and these plaques are nasty. They can but but this no one I don't I think this this article in and of itself is a demonstration that man you got to really think outside the box. I don't think anyone could have predicted that treating one's liver and it seems like in this case you're arming monocytes to go mop up the ApoE or whatever sort of proteins might be causing the the plaques in the brain but it's sounds like the monocytes are getting armed within the liver and then they're trafficking their way into the brain to kind of become this defense mechanism for preventing it. But that this is still very early on John a lot more to go on but it's it's really I want to hammer home the the thought of like just how um amazing it is that you know uh someday we're it's and by the way tr treating somebody with drugs that try to get the across the blood brain barriers is obviously a challenge. There's they've had a lot of um drugs you try to label with something to have for active uptake but that become becomes very problematic again because of that barrier. And then some drugs are given intra intracranial cranially you can imagine how that isn't really a sustainable way to treat people because you know if you're in an early stage the last thing you want to do is maybe tap into someone's brain but man if you could just give someone a shot and augment their liver and they can treat Alzheimer's man that that's a solution right John like that sounds good. And uh and then I I'll round up by the hey stay tuned on the the the the let's call them uh in vivo um vector based um car or even therapies right so the G and V space has really got some impact and not like not like arming T cells and things like that not the not the like the ex vivo autologous therapies but more the in vivo vector uh based therapies could be very beneficial in the Alzheimer space but this I I would I would invest in this John that's I'm thinking this has high upside if you could start unlocking how the liver because the liver is a very easy uh organ to reach and and turn on so to speak but fascinating stuff John so um we we we are we gonna do one more yeah yeah let's let's move on to the main topic I think um I think it makes sense um so I'll I'll introduce it and we'll see where it goes from here.

SPEAKER_02 26:30

So obviously you know we've been a business that's been going for over 40 years and and um and w we're slowly expanding our services to meet the needs of the evolving you know evolving uh requests from from the pharmaceutical industry so we thought maybe just worth touching on um we're giving it a title recent trends in bioanalysis the evolution of client needs instrumentation platforms and regulatory requirements because you know if you look even if you go into the lab today the instrumentation is is very different from where it was you know 10 20 years ago because you you you the the the therapies have become more and more complex and so the analytical methodologies to deal with those has become more and more stringent um and it's not just about complexity it's you know we we want to go and look at lower lower levels um it yeah the the the the challenge the challenges never stop I mean we we we can't sort of rest in our laurels as a bioanalytical lab because the needs are always evolving so we always have to do what we can to meet those needs and and you know um ensure that we we can help you know in drug development.

SPEAKER_00 27:48

Yeah no and and John I I I definitely think that this exciting topic right all these one of the best things about working at a CRO is you do get to have a good pulse on trends and trends come in the form of uh you know what latest and greatest drug and drug modalities the instrumentation platforms that we get to deal with regulatory requirements and in general our client needs right so we've you know we've both been I've been here 13 years at K Cast Bio over that you've you're you're rolling up on five I think right and we've had a nice um it's been wonderful to see the evolution and and that brings us to maybe even to the first topic here right like it's we always know that in bioanalysis there's often this need for just expanding the sensitivity John right and I'll maybe I'll let you go first because I think in the mass spec world even in my term time here I I can I I remember when we got our first um like 5000 instrument and that was a big breakthrough but now you can go ahead and tell us more about how how advanced Scieks and that's a Sciex yeah well yeah I mean obviously we're we're a SciX shop in common with many and and the they they've had an interest in evolution over the years.

SPEAKER_02 29:00

They're there there are early instruments with the API 3 which in terms of sensitivity etc and this I'm talking going back um god 30 odd years and um the the API 3 plus was was a was an excellent instrument in terms of sensitivity and none of the other manufacturers could match it. Then they we moved to 300 365 which wasn't a great instrument it gave the other manufacturers a chance but in terms of uh SciEX have done a lot of work in terms of um looking at the the and the other manufacturers too is how can you better improve the sampling from the ionization process um so we then went to the 3000 which was a step ahead a quantum leap from 3000 to 4000 5000 was an improvement up to the 4000 in terms of sensitivity and then we had another good jump from the the the there was the 5500 which didn't make a great deal of difference but then 6500 and 6500 plus was another big step up and and the needs of our clients were always pushing for these these low levels of drugs and in in in plasma and other other matrices. Now we're at the API 7500 and 7500 plus and it's interesting you talk about you the you see the the API 5000 for us that's now a platform that is being retired because the because we we need with particularly with large molecules you need those higher performing platforms. So we're we're uh almost exclusively our fleet has moved to 6500 and 7500s because obviously you can you can you can redevelop or revalidate uh and and show that assays that worked on a older platform can now be easy readily run on a a newer platform like the 6500 and 7500 but you're never going to an assay that you have running on those two platforms you're not gonna get running on the five thousands and lower so although those were great instruments their their time has come and we're we're much more in the the 6500 and 7500 fleet because that's where you need to be to and it's it's it's a combination things it's it's actually looking for the sensitivity that's needed for for getting down to low levels of these these um drugs that we're being asked to investigate but it's also trying to um minimize your consumption of samples so it's like you know for us I remember the early days again when we're at API 3 etc we were routinely extracting 200 microlitres a sample or half a mil of sample or more than that. Now we're at you know 25 to 100 and 100 is is is is a large aliquot to be using we're much more than that 25 to 50 because the improvement in the instrumentation it means that you're not consuming as much so you don't need to take as much and it also then helps in terms of you know construction of animal studies you can maybe take cereal samples from animals as opposed to you know um you know a lot more sparse sampling so you you involve more sample more animals and that means more more dispos animals you know is killed at the end of a study things like that so that there's it's a lot interplays so the sensitivity of the of the instruments can have like a huge amount of benefits to how the whole how how the whole sort of process works.

SPEAKER_00 32:34

Yeah.

SPEAKER_02 32:34

No and and um you know uh as a couple things uh I was John you mentioned our 5000 suite there um I'm we had um a bunch of new 7500s and pluses uh arrived today and by September our 5000 is gonna be gone I was on site yes yes 5000 they're gonna be gone and then there's even an an 8500 right but that's a high res instrument yeah that's yeah I think it's the 8600 but oh sorry the 8600 I think it's like a I don't know too much about it because it's I think it's more of a time of flight based high res so different from the from the orbitraps that we we we've typically worked with but um yeah it I again there's it's there's a lot of attention being paid to the 8600 and what it might offer um because of course you have to keep abreast of of the developments and and what what these instruments can offer you in terms of you know you've got a a an assay that might be a struggle on one platform but you know you come at it from a different angle that you might then have a solution.

SPEAKER_00 33:36

No I I think it's a just even as I sit here what a it's I'm fascinating that I've been working here long enough to see at the time I joined the 5,000 was like it was on the market but it was a big investment for us. True big step up from the 3,000 4000 instruments and now here we are it's they're being obsoleted because they're not sensitive enough right and I think we're of the val of the 40ish instruments we have I think we're gonna have something like 14 that are 6500 and the 26 of them that are going to be in the 7500 or higher range. So we're really I think it's in I hope our audience hears us we are state of the art when it comes to mass specs these days but uh John maybe switching over to the other like so on your side you've got the sensitivity with the instruments and then on the ligand binding assay space or in general maybe large molecule biomarker space the the latest and greatest instrument is or platform is the the new LISA system which is from a company called Alomar and they sell an instrument called the Argo and this this is really um you know you touched on sample volume one of the first things right out of the gate that makes this instrument incredibly attractive is is the is the ability to use like 25 microliters um per sample and it requires almost no dilution of the sample and so if you if you you know that that the ability to use a small quantity and get uh readouts they they have some plexus that go up and I think they're launching one in the 350 range but but the technology itself the advancements of the technology come from you're still using antibody pairs and one of the antibodies is biotinylated uh but the and and then and the other one is is a polyatyl so they're using like PCR for the readout right so this is the the instrument itself has uh it's like a big it's like a qpcr box and then when you get up into the higher panels you actually have to um load the instrument up and then move it over to a next gen sequencer but but the reason why this gives you this high sensitivity and even specificity and selectivity and everything is there it's like an amino PCR reaction now so it's kind of amino PCR was a technology that's been around for a while but it never really worked well because it was challenging because you you didn't have an instrument or anything it was kind of you kind of had to hodgepodge your way through it maybe do an upfront amino assay and then kind of try to adapt it to some sort of PCR instrument where the people at Alomar have capitalized on that and they're taking antibody pairs that normally detect in the nanogram level and they're getting sub sub picogram femtogram or even down to allomolar uh ranges of detection and and this has become it's it's really taken off for us. We've had it for over a year um there they're the LMR is really uh looking to partner with us on a number of their kind of single plexes some of their homebrew kids a lot of it's focused on the neurodegenerative space there uh we're looking at doing some fit for purpose validations of their five plex which is interesting enough it's got Ape OE on it and uh you know things like NFL and uh Tau and brain derived uh sort of uh biomarkers as well but the the the fact that it can take you know commercially available antibodies and make this incredibly sensitive uh method out of it is really I don't know if it's revolutionizing the biomarker field but it has huge upside for biomarker analysis where you know not only are you not having to do any you know dilution corrections so if I'm measuring a spattering of biomarkers with a one to four dilution and a bunch of other at one to ten that that can create some everything's done at the same dilution that's huge has a very large dynamic range to it so that you can get detectable readings on just about every sample. So meaning you know BQLs don't really help a scientist that much this they've overcome a lot of that. Yeah and it's just it's a wonderful platform. And then I'll I'll say it's we're evaluating it for PK and we're starting there was a publication out showing that there was some advantages to doing it by PK and PK has huge upside because again same thing there if you're doing different doses right dose escalation you almost got to create two curves sometimes right John because it's so vastly different.

SPEAKER_01 38:05

Sure.

SPEAKER_00 38:06

And in this instance you're gonna and that dilution itself I mean statisticians can overcome some of that but the data's a lot cleaner if you could use one dilution you don't have to pool animals because you can use low volumes to get the answers. And so that we're you know stay tuned but we've seen some sponsor success we've got a sponsor that we're looking at doing this and it's showing real promising like we adapted an MSD method and it was in the I think the mid-picogram like 50 to 100 picogram level and now we're um seeing it well below the picogram detection level and this is our sponsor very interested and then we are working on a generic assay which you know if you're familiar with what I mean by that it's measuring a some sort of monoclonal antibody or any sort of antibody based conjugate where you you don't need any special reagents and you get high sensitive methods. And you can, you know we have generic or universal assays by ligand binding assay and then we have some by hybrid mass spec, right, John, where you follow the the digested peptide so I I think when it comes to like trends in bioanalysis sensitivity is always going to be on there John and uh as we work here at KCAS bio we're I think we're right on top of it with our investments in in mass specs as well as in the NeLISA system.

SPEAKER_02 39:22

Yeah absolutely yeah yeah and I know I know I've been in discussions you know where you talked about some of the Alzheimer's or the neurological biomarkers and and the the the the levels that people were wanting to measure didn't seem feasible whereas now you're talking it's much more much more available to you in terms of the tick what the technology is giving you.

SPEAKER_00 39:46

That have meaningful data and uh you know it's there's there's just huge advancements in being able to not have to sample CSF but still be able to and and it can be a good diagnostic tool as well right to start looking at and then one of the projects we hopefully will be working on is like we got a um if you looked at healthies cognitively impaired in true AD patients that would be a good thing to be able to discern to sort of start you know the cognitive impaired it means people are kind of progressing but if you get biomarker panels I mean I don't know about you John be real nice to know oh do I want to know I don't know I'm kind of torn by that now but it would be really good to if you're predisposed or any of those things it'd be good to be able to really start to understand um it's always if you can diagnose a disease earlier you always have a better prognosis and that's somewhere where the NOLISA could really start to tease out and not having to take CSF samples where if I could envision really large longitudinal studies that can help you really understand that and really arm physicians with taking a plasma sample and being able to help those individuals that are worried about it, right, John uh so anyhow my two cents there.

SPEAKER_02 41:02

So there's your high sensitivity piece. Yeah so the other other side of the coin for us is then um high selectivity um I again improvement in the instrumentation is made a lot of it made a oh it made a lot of difference um you know for us it's you know in the on the mass spec side of things we we're not we're bu steadily building a fleet of high res mass specs um I mean I've I've had Isays in the past where um a triple quadruple just couldn't just could not filter out enough background to allow you to to measure your analyte of interest um we're getting we we'll touch more on oligonucleotides later but we found that again you know the having high res instruments um for oligos just just it it helps in terms of reducing development time yeah you're you're cutting out background um because that that's really where the selectivity becomes key your sensitivity you actually gain sensitivity from the selectivity because your single signal to noise is is the is the low and and so we can we can tease out data from you know from background the having that wide range of platforms between the triple quads and the and the high res mass spec is really it it it greatly enhances the suite of what you can do for your for your customers. So you you need that whole whole plat whole set of platforms to really address um you know the the the assay needs.

SPEAKER_00 42:41

Yeah and and John so on the Ling and binding assay side we have a variety of platforms that are highly selective but one that's um somewhat new to us although I think it's been around a while is the Alpha Eliza and and it's a little underutilized I think but we are finding more and more sponsors that run it in-house because it has great selectivity to it and how it works is you just again it always starts with the critical reagents you know those are near and dear to me but you've got an antibody pair and you know in a conventional ELISA you might capture with something and then detect with the other and then in an MSD format or even Samoas and you you end up using some sort of uh um you know biotin kind of streptavidin kind of helps get more capture antibody better detection there but on the alpha Eliza it's all it's all um in liquid phase and what they use is uh they call it like an alpha donor bead and you you biotinylate the antibody onto this bead but this bead when you hit it with a laser you can get excited then they have this alpha Lysa acceptor bead and on that is the is the um you know the technically the detecting antibody and so when the two of them get near that when they bind to each other and you get the analyte of interest and you've got the capture antibody And bead on there and the detection antibody and bead on there. You hit it with a 680 absorption or laser, and it and then what it does is because of that proximity, it emits on that like acceptor bead and it gives you great signal. It's they're tough methods to optimize, John. And it's a service that um what we're finding out is it it really it's it's I don't call it an oldie but goody because I do think Alpha Eliza technology has been around for a while. It's something we've been offering for almost a year and a half or two years now, and I but I do think it, you know, it might not have quite the sexiness of a high-res mass spec and some of what's going on over there, but I I think it's important to mention how and how this is as we begin to think about trends in bioanalysis, the evolution of our clients' needs, as well as like the instruments and things that we need, this becomes an it's actually another tool in our tool chest. And you know, I might not start with an Alpha ELISA program, but to those that are out there using Alpha ELISA that need CROs, we're now got a couple of these under our belt, and we really feel like it's something we can uh help you out with. But it it also could be when you're at your last kind of at your last straw, it's another platform to go and look at to provide good quality data. So I don't know, John, if you had anything to add, but that's it's really a good idea.

SPEAKER_02 45:11

I think it's I think it's that I think it's that whole thing. Yeah, we talk about high-res mass spec being sexy or whatever, but ultimately you just want as many different tools available because if where one might fall down, you want to have something else that might be able to step in and and take take on your problem and solve it. So yeah, it it it's it yeah, it it like the h high res is obviously that's your big investment instrument at the moment. But again, that may change with time, as you know, we we we see with all instrument all all mass specs anyway, they're very expensive on initial launch, but as they become more accepted and the the the cost goes down. Um but who knows what might be the you know the next improvement uh in mass spectrometry uh quantitation downstream from here, you know. So we we we talked selectivity, we talked sensitivity. Um you we we as we talked about trends and bioanalysis, this is something I don't think about at all. Um, but is there's a lot of discussion in this on your side of the the business in terms of uh single replica analysis, and I remember um you know being at EBF that must be I don't several years ago, and it was like afternoon workshops, and um, you know, when you're at a conference, you you like to sort of dip your toe in other other um areas and see what what has been spoken about, and this was my first inkling about you know the the norm for multi-replica analysis uh with it with within you know leg and binding assays where for us it's just you do one measurement and that's it. So I'll pass it back to you because I just want to at least give a bit of my background to it, and and like I say, I it's not something that affects me, but I know on the on the um like you say ling and binding side, it's it's a hot topic.

SPEAKER_00 47:05

Yeah, no, it John, it's it's fascinating that you've never had any replicate sort of issues on the mass spec side. I guess I kind of knew that, but you reminded me that. And in our world, you know, I've been doing I feel like God feel like I've been doing this so long. Everything was triplicate at one point.

SPEAKER_01 47:20

Yeah, yeah.

SPEAKER_00 47:21

Right. And then even as I got into the CRO space some 18 years ago, we almost never run triplicates because of cost, right? It was always just duplicates. And and then there are but there are still some methods where it makes sense to maybe prove that you can go from some cell-based things or activity assays, you might even do four replicates and then triplicate and down to duplicate. But the norm is duplicate, and you you touched on it. It was actually, I think, two years ago at EBF, it was a big section of the afternoon. And so, you know, the PCR space, I'll say that they've been doing singlicit for a long time too, John. Right? They don't do a lot of replicates there, and those are even like you know, IVD approved tests that are a single rep, and it's pretty physicians rely on it, but it's been kind of like taboo in the LBA space, just just because of like the the the um I don't know the the stress as an as a when I was an analyst, I would have hated running single single kits. It just gives us more confidence to be able to do duplicates, and and I hope you can kind of hear it in me, John. I was I think if we had if you had had this discussion with me, maybe even after EBF, I wasn't sold after them, but the field is really pushing this. Yeah, as I'm getting, I'm kind of warming up to it. I'm getting I kind of I hope I don't sound like I'm stodgy or too conservative or anything like that. It's just I try to look at all angles, right? Not just saving on money and things like that, because although I I would say that's going from duplicate to singlicate isn't gonna save you 50%, John, right? It's gonna have some, you're gonna get some savings. And we put it at it, we are offering it now regularly. We have some projects where we're doing singlicate replicates, and it's maybe like a 20 to 30 percent saving, which is significant, and it also reduces footprint, right? You know, using less materials, all good things. But I think I think I I still want to put a little bit of caution around this, in that you really got to prove that the singlicate replicates are there.

SPEAKER_02 49:19

Yeah, I think I think for us as as bioanalytical scientists, you want to do the right things. You don't necessarily you don't want to jump to an alternative approach until you've really proven that it's that it is safe. Yeah, I think for want of a better word.

SPEAKER_00 49:35

Yeah, well, I I think there are some people who are you know maybe a little bit more risky and they're convinced right out of the gate that singlicate can be done. But I think in terms of regulatory submissions, I I think if you can prove it during development and validation and um you want to go into your sample analysis at Singlicit, I think you'd be okay. But again, you got to demonstrate it. It's not just like I'm doing my development and validation and single replicates and calling it good. You still run across um, you know, the the there's an argument that if you do singlicit for bioanalysis or you know, sample analysis and you get a result that's um just you know spurious failed, whatever it might be. Do you have to rerun the whole plate, John? That becomes the issue there. And then you can counter it and say, well, if you were running duplicate replicates, there's a good chance even if one of the two were out, you still got to repeat that. But in my world, if you got two of them, if they're if you can do a grub test on there, you can kind of confirm through stats that one of them was plate washer got blocked, the technician had an error, you know, something like that. But you know, the world seems to be pretty convinced that you can do singlicate. And um, you know, we'll we'll have some case studies out here soon, John. But I do think it's I do feel like it's gonna be the wave of of ligand binding assays, but you know, I think it's gonna be very still gonna be somewhat method dependent. I I don't know how quickly it will adapt to PK and ADA, but in the biomarker space for exploratory biomarkers, I think it's really becoming more the default, would be good. It's not there yet, but I think in time it will. And then I think slowly but surely you'll see it in the PK and ADA space. But it is it's somewhat new to us here, John. We've been well, you know, it's not something we would routinely be offering, even though we talked about things from a couple years ago. But you know, we we want to do what's as you said, we want to do what's best for our clients as well as the individual's work doing the work, right? That there's a little stress there when you go down a singlicate. But I think you get confidence, right? As soon as you if you're an analyst and you're trained on the method, you should be fine with running it in singlicate replicates. But it is is a hot topic in biological.

SPEAKER_02 51:43

So so do you routinely do ISR and you know, encourage sample reproducibility? Um as part of so you're doing that, because again, that's our that's our defense um for showing that single kit is fine. Because I mean, obviously when I started, we weren't doing ISR, and there was a lot of you know a lot of discussion back and forth, and then it became you know, it be it became the the norm for us to be doing in card sample reproducibility in you know talks and and clinical studies. And and I've I obviously we've I've had assays that they weren't functioning well and and we would never have known unless we did the you know the ISR that that pointed out that there was a problem. Um and I it has made I in my opinion it's made things much more more robust. So you you that so although you're doing it that you still would like to do the duplicate.

SPEAKER_00 52:40

So it's so you bring up a good point about PK. There's no ISRs for ADA and biomarkers.

SPEAKER_02 52:45

Okay, yeah, sure, absolutely for PK.

SPEAKER_00 52:47

If you do have a result that is an outlier, maybe a mistake, whatever it might be, you can come back and retest that as part of the ISR batch. You're absolutely right. But in the you know, if I'm doing a biomarker 220 plex, man, and I got one of those that I'm playing. I don't know. That's a very expensive retest. So but there's ways to must, you know, it's hard to address exactly what you do there. Um, although I think if you've got the rest of the time points, you can you can maybe go without it, or there's some some workarounds, but you bring up a really good point about ISRs reinforcing the PK side of it, but they're not done.

SPEAKER_02 53:26

True.

SPEAKER_00 53:26

And and um but there are some people who want to do them. Um for biomarkers are there's a lot of resistance from many people, including myself, that would tell you it's not a lot of value in doing um ISRs on ADEs because you are doing the three tiers, although if we're going one tier, maybe there might be. And then the biomarker space, there's there is there is like a small group that's always opining about wanting to do um, you know, they don't want to call it an ISR because we don't truly have like a standard and things like that.

SPEAKER_01 53:59

Sure, yep.

SPEAKER_00 54:00

It's just more like a uh an incurred reanalysis.

SPEAKER_02 54:04

It does become but is a re a reproducibility of measurement so long as the because the you because your samples are snapshot in time, you can know that snapshot is consistent anytime you go back to it.

SPEAKER_00 54:15

Some LTS does some of that. I don't know, there's some arguments around how to do that, but there's you know, it's well I'm not gonna I'm certainly not here to promote the other when it comes to doing ISRs on biomarkers and things like that. But uh I like what you said about reinforcing the PK data through ISRs is a great way to kind of check yourself, make sure you got it doing it right.

SPEAKER_02 54:35

Yeah, and and and when you talk about no measurement of 220 biomarkers, that's not something I have to ever have to worry about on the mass spec side of things. You know, I'm it's it's it's not very often we're going for many analytics, you know.

SPEAKER_00 54:48

Um that's the that's the trend. That's another trend there. People are you know, biomarker high throughput panels there are never does they're not designed to test thousands of samples.

SPEAKER_01 54:58

Sure.

SPEAKER_00 54:59

Yeah, but people, but there are people who are starting to look at it the other way around, which is interesting. I I don't know. That makes sense. Some people want to they're coming to us and like, hey, we we got 500 samples, we want to do a retrospective study. Can we do them all on this, you know, 220 plex or whatnot? And we're kind of like it's better to maybe take a pilot sample, take a spattering of them, some sets of them, take it a look at, fill in a plate up, and then go from there, see if it's worthy of it. Yeah, and even then, after you get some hints, I would say, why am I testing 220 analytes? Maybe there's only five, seven, ten in there. I would want to run my other 500 or 5,000 samples on. So there's there is some strategy involved there when it comes to maybe biomarkers in general. But okay, John, we're we're moving right along, right? We got a couple more topics we want to touch on. Sure.

SPEAKER_02 55:45

Certainly near this is mainly you because uh I think that's well, I suppose it's both of it. It's like this is again a lot of discussion in the um in the bioanalytical field, and and a lot of this was again at EBF in Europe have really been driving this. Is that is the use of fit-for-purpose validations and biomarker asses. I mean, I've I've I've sat in a lot of sessions regarding this in in Barcelona, and and so I I mean it's and actually we're seeing it more in terms of you know talking to to clients, you know, that it used to be that we get on calls and and they'd say we want to validate this biomarker as I say, and it's like, but do you really need to be a lot to and fro? And I find that it's a bit it's but everyone's much more calibrated in terms of what their what their expectation is, and they are just talking, doing you know, uh uh precision accuracy run, limited stability, whatever. And and the ex and it was interesting to have a call earlier this week that we were both in where it was no these these two biomarkers are absolutely primary uh prime a primary data point that we need, these have to be validated, but that's an unusual request nowadays.

SPEAKER_00 57:00

Yeah, no, um, so it's it's a little bit to unpack, right? Because a lot of people use terms like intended use, context of use, fit for purpose. And I I think sometimes people just want to say, if you slap a fit-for-purpose validation, you can do whatever you want. That's just not a true statement, John. I want to be making sure that we understand that um, you know, when you do the a fit for purpose validation can have many, many flavors to it, but it still has to be driven by what are we doing with the data? Um, what do we have that we can do to make the method robust, um specific, uh sensitive enough. So there's a lot of things that go into play when you talk about intended use and context of use. And that's a whole nother I think that's a whole separate podcast.

SPEAKER_02 57:52

Sure, absolutely. Yep.

SPEAKER_00 57:54

But but I think for what I'll say about this why why why is it such a trend suddenly for fit for purpose validation? It's because there's a war on the word qualification, John, right? I think it's created a lot of confusion because I feel like back in 2010 there's some guidelines that came out that made it very clear around what your fit for purpose was, but they never talked about qualification versus validation. So what's tripping up our industries is in our world, John, our bioanalytical world, if someone said they've got a qualified method, you and I don't think that it's a bad method that couldn't meet performance characteristics. We just assume, hey, the the use of this was they didn't need a QA review. Yeah, qualification was good enough for their uses. But if you're a CMC person, if you're if you're running a qualified method, that means it failed validation, John.

SPEAKER_02 58:47

Oh, really? Okay.

SPEAKER_00 58:48

Yeah, that's where the confusion lies. When when you've got people who are uh, you know, if I if I say to someone, oh, you hey, we need a validated method, but if I'm talking to somebody who might be more of a CMC GMP expert, who's now maybe finds themselves needing to do bioanalysis, if I say, hey, we're gonna qualify the method, that that they take that as like, oh my God, what happened? Oh my god, it's gone disastrously wrong. Sure. Because you know, in the again, in this in the CMC space or GMP space, that's that's how they define something. When you're qualifying a method, it means that it failed validation. And by the way, it still gets a QA review. Now, in our world, the the term qualified means it lacks the QA, but everything everything we do is done in a GLP environment. It's still done in a similar fashion, it's just not having that back end Q it QA review.

SPEAKER_02 59:40

So now and it's also you know, it's also um it's it's basically showing that the assay as it's developed is suitable for use to meet the needs of what the client wants for that particular study. Yeah, that that's where we're that's where we're going with qualification.

SPEAKER_00 59:59

Yeah, I mean, I think uh to me, a qualified method in our world could have percent CVs, percent recovery, all less than 10 single-digit levels would fly through a validation. It just didn't, because of the intended use for yeah, and by the way, you can submit you can submit data in any sort of submission to a regulatory body that doesn't go through a QA review if you've a priori your data that way, right? You know, so that you've set it up the right way. But um, you know, back to the way we're kind of looking at it now, uh, because the world is see, I think there's a lot of members of EBF and there's a lot of members of um the APS community and uh even the WRIB communities, where they're just like, okay, we're gonna call everything fit for purpose validated, but nobody's really addressed that regulatory rigor. So I think it's still I still think this is still in flux something that might trip us up. So we're starting to talk about it in terms of like a regulated fit for purpose validation or a non-regulated fit for purpose validation. But some people don't like that because if it's fit for its purpose, and that purpose is three A and P runs with no qualification, no plan, no report, right? No, sorry, three runs, no, no formal qual, no formal validation report, no formal validation plan, and no QA review, that can still be a fit for purpose validated method. Yep. But that gets that gets hard. It's gonna create a lot of confusion. And and what the last thing we want to do is be deceptive and say, hey, this this we validated this, this is a fit-for-purpose validation, and all it is is one A and Q batch, one A and P batch, John, right? Yep. So that and and then you know we've started thinking about tiers. If we we do have a bunch of tiers, but there's a lot of people that are I'm starting to hear about oh, these tiers are silly and this and that. Well, if all I have to say to some people is if you keep hating on things, please come up with a solution. Yeah, because just just telling me that everything can be fit for purpose validated, and you just it's a blank canvas, that that right back to where I started with. Don't just slap a fit for purpose title and do whatever you want. That's bad science.

SPEAKER_02 1:02:06

That's no, no, you yeah, you have to you have to have agreement between you and your your customer that what you're suggesting for that fit for purpose validation makes sense to both us and them.

SPEAKER_00 1:02:17

Well, I mean, even think about the reviewer, right? How about the the person who's reviewing the submission? If you just put fit for purpose validation on it and you don't put any caveats around that, and then I start looking at your data on all you did with one AMP run, I'm gonna think that they know what they're doing. Yeah, but then maybe you have a fit-for-purpose validation that's got six AMP runs, you put a plan report and it's got, I don't know, 180 pages document for its validation report, but you're gonna call it the same title. So I still think there's some things to be ironed out there. And by the way, I sit on, I'm involved in a whole bunch of communities where we're trying to hash this stuff out. And the problem with that is it's an echo chamber, and there's maybe within it just a if if if we just let a small community of people dictate things, it's gonna be problematic because there's a whole bunch of people. John, remember if you're talking to a European client in general, if you use uh if you're in a bioanalytic bioanalysis setting and you use the word qualification, they're gonna think you that you don't know what you're doing. But regularly here, we take calls all the time from people that are very, very good in translational medicine translational sciences that say, hey, I need a qualified method. It's still to be determined on just where we're gonna land. Although there was a publication back in March that said you should avoid using the word qualification. So all right, I think I beat that one to a dead horse. But I got opinions, John. I got opinions on that.

SPEAKER_02 1:03:39

Let's move let's move on. Final like final topic, because I think this this really this is this is indicative of of why having this suite of instrumentation available is key. And um, it's what we're seeing. I mean, probably, you know, I always as a mass spec person, um, we had very little contact with oligonucleotides. Um I think the first time I ever worked with any was actually on the time of flight my days at Kratos. But when actually when it came to a bioanalytical lab, um we there was no real discussion of of of LCMS of oligo oligos because they're tough analytes. There's a lot of problems with adsorption, carryover, um, all sorts of stuff. Chromatography could be temperamental, um, you know, just just a lot of reasons why um you know we just weren't we weren't getting any requests as as there's been more and more emphasis on oligonucleotides, both in terms of just straight allegos or sir and a s or it it it arcs where you got you know the antibody with an SIRNA. Um the the my spec has become more integral to to um you know it's it's a an a more integral tool with everything else that was already being used in terms of giving answers for for the these oligonucleotide type of of analytes. Um again for us it's w we're uh having that full toolbox of triples and uh high res instruments is is vital to in terms of of meeting the needs of oligonucleotide analysis. And even I was having a discussion yesterday with Rashnor about you know what they do on triples, and although the API 6500 is a is a lesser sensitivity instrument than the API 7500, what they find for some oligonucleotides is because the the signal to noise on the 6500 is less than the seventy five hundred because the smaller orifice up front, they can actually get better performance for some oligos versus seven seventy five hundred, and then they can get obviously a uh complementary performance on the high res instrument so that it gets it's that variety of tools. But I didn't want to get too far down the mass specs. I'll pass it back to you. And because we're we're using a lot of tools for allego measurement.

SPEAKER_00 1:06:05

Yeah, it just goes to show you you can't you can't be absolute when you're talking about the sensitivity of each of those instruments. You've got to be careful because there are those caveats, right? But you know, y in terms of oligos, one of the things that I love working here and many, many reasons, but we got we got many tools, John, right? You've got mass spec, you've got high-res mass spec if needed. We have hybrid Eliza, and I'll touch on those in a minute.

SPEAKER_01 1:06:31

Yep.

SPEAKER_00 1:06:31

Very powerful tools. PCR is the fourth one that we offer. And then there's like a hybrid hybridization LCMS, right? Which we haven't quite tried yet, but I think that's in there. Yeah, so so we have those four buckets, but John, so maybe not surprising at all. The the PCR is the most sensitive, right? Yep. So um actually a a client, uh a pretent a prospective client asked us after we went over our, you know, we have a slide showing all four and the ranges, and I don't have it memorized what the ranges are. But um, they asked, hey, you know, um what what what would you recommend to start with? And that that's a tough one too, John, because you gotta kind of if I if if if you got to use the tools that you have, not everybody's got all those things like we have right. So that to me, it's like I would start with whatever I've got that gives me the answer I need. And putting that aside, right? You know, if if the if if you don't so so maybe you can do it by mass spec, but you know, we do have some case studies that show both the hybrid LISA and the PCR can get maybe two to five log more sensitive than whatever we were doing by mass spec. We were using high-res at the time. So so to me, it's like the real like uh the way I answer this is if if you if you have if you came to us today, we would probably default to a PCR reaction, John, unless there was some other reason. And and or we would look at what are you more accustomed to that might be beneficial. But it all the devil's in the details around how sensitive you need to be and that. But and then it it's challenging because are you talking about some sort of arc, right? Where there are huge advantages to doing that by mass spec because you're you're able to do a one and a half plex, or I think we've got a one and three-quarterplex, right? We got some terms, but that that becomes an advantage. But I do think when it comes to Oligos, it it's great to have so many tools at one exp one's disposal. And um PCR is a tried and true um uh platform for it. I'll say about hybrid Eliza, this has come a long way in the three to four years that we've been offering it. And what I mean by that is so you still gotta make the same probes, it doesn't matter the platform, you still gotta go and make a probe, whether it's PCR, high-res, right? You still got to go out and make some sort of nucleotide probe, right? So that's still a critical reagent that all of all of the platforms need. But the hybrid Eliza, because of some of the more commercially available reagents with the dig detector is what's used there, the ligation sort of reagents that are used. We're finding that over the last few years, we're we're we're getting much more success because people are doing more and more of it. So, like any capitalistic market, there are more and more vendors providing it, and the quality of those reagents has improved. So hybrid alias is a really good platform.

SPEAKER_02 1:09:20

And I I think that's that's a key point, and I I I've returned to this multiple times. We're lucky that as end users, we have you know suppliers in for on the mass spec side of things, and who are always looking for improvement too. So for Olegos, I talked about com uh temperamental chromatography, but the column technology is improving where they're actually developing phases to specifically deal with it with these types of analytes, and so you know, we're really benefiting from a from a very sort of I mean obviously it's commercially driven, but a sort of collaborative uh supplier set who who are looking for tools that that will help us and in turn help our customers.

SPEAKER_00 1:10:07

Yeah, no, definitely. And then one more thought that flew into my head is if there's any metabolites of your uh oligo, then mass spec becomes very attractive. If you look at the activity of each of them, that becomes huge.

SPEAKER_02 1:10:20

And and for high-res that become I mean that that high-res really steps into its own on that. Yeah, so because we can do full scan acquisition, and you know, you can actually then go back and um interrogate that data later to look for for metabolites, etc. So yes, mass spec can can certainly do a lot in terms of that piece.

SPEAKER_00 1:10:42

Only if you need it, right? Because that was the other factor that I want to be clear about when you when we're starting to assess the first platform, is if you if you know right at the gate that you got a metabolite that you need to deal with that has some activity to it, then you just you just gotta go right to the more high-res or mass spec, right? Um but but if if not, then then I still think PCR, although I think PCR has some ability to do that too, if you design it the right way. Okay, John. I think we're wrapping up, right?

SPEAKER_02 1:11:09

I think we probably are. I think we we I wasn't I think we've talked a lot on this, wasn't quite sure where it would go, but this thing this has been quite quite satisfying. So yes, I think let's um like I say, this is i it's always it's always fun see the developments and what we have to do to to address them, and no doubt the challenges will keep coming.

SPEAKER_00 1:11:28

Yeah, no, it it'll the moment you think you uh you think you got a grasp on it, it'll change, John. It's it's it's I can say if you don't like constant changing and so forth in the CRO space isn't the right one for actually I don't know if drug development in general is, but it is it's wonderful to you know think about what we just discussed right now a year or two ago would have sounded very differently, right? Yeah, absolutely. So much that's been out there. But let's uh let's do some quick uh we got weekend plans going on, John.

SPEAKER_02 1:11:57

Um nothing much. Um I I I I got a weekend trip which I have to do. Um and um I sort of probably end up I'm gonna be in Boston, I'm gonna link up with my eldest son, but I have other things I have to do around that. Um yeah, it's just not much on the weekend. Sorry, sorry.

SPEAKER_00 1:12:17

Is that is that part because it really uh you got any fourth of July plans? Are you going to bought what you're doing?

SPEAKER_02 1:12:21

Well, see, Fourth of July, I mean I'm quite happy to take the date, the time off, but ultimately I'm the reason you're having four Fourth of July, so it's not something we particularly s register. It's it's a it's an American holiday, and I'm obviously I'm British. I love having the time off, you know. It's like but but July for us is a big month because it's like there's multiple family, you know, multiple members of the family have birthdays in July, and then I'm actually going back to the UK late in July, and I've got much more planned for that. Um to go up a music festival in Leeds for a day, and there's a there's a site called Great British Car Journey where you can pay to drive like old cars from the 50s, 60s, and 70s that I'm doing. Um and so that that's what and then it's just you know going to some of the I've got a few extra days just visiting other small towns. I'm staying in Yorkshire. There's um I'm ho I'm hoping I can fit in the Keith Leonworth Valley Railway, which is uh uh you know an old steam steam. They they they basically do trips between uh two towns, and it it's either steam, steam train or uh or a diesel, depending on the timing. So there's there's like lots of little things I'm trying to do, it's just a nice trip, but it was actually the music festival was what triggered it. It's a band called Sunshine Underground, who I saw their last concert in Leeds several years ago, and that was that supposedly them done, and then they sort of they announced they're doing this one-day music festival in Leeds, and they're supported by um Future Heads, White Lies, Hard Fi, and another band who I haven't heard of, but there was enough bands there that I hadn't seen, and I'd quite like to see them. I thought I'm going to do that, so that's why I'm back in England in late July. All great stuff, John.

SPEAKER_00 1:14:10

We couldn't be further different from what we think of as vacations, but that's okay. I love that you're doing all like what a nostalgic trip driving old cars and taking steam trains and then going to things like I I so I've got a couple of days in York, which is a fantastic city.

SPEAKER_02 1:14:25

If you ever go to the UK, always do York. But you can get you can actually in York you can walk the city walls because it still has intact walls round the city, yeah. And you can, you know, you they've got a few gates round the and you go up the stairs on the gates, get on the wall, and just wander around. It's and York Minster is just a fantastic cathedral, and so yeah, it's it's just a lovely, lovely city. So it's just I haven't been there for uh a long time, so it's just been nice to catch up on some of the stuff.

SPEAKER_00 1:14:53

Yeah, and I I I don't know, John, you've been working with me long enough to know I like to try to blow up the state of Nebraska with live fireworks, but we got a curveball thrown at us as you know uh traditions are wonderful, but um my my son has decided to depart way. We're we're not gonna spend the 4th of July with him, which is fine. He's a grown man these days, he's 20. Um, but he's he's going down to the Ozarks for some fun and sun with the boating and the you know, good time down there. We'll we'll we will go to Nebraska, but won't just won't be a little different with in terms of uh not having him to help light the fireworks off. We'll see just how big the display is this year, but it'll still be a fun time. I do enjoy um it's a lot of fun, and then we do have a little bit of a family reunion with her, and one of them lives on a little cove, so we'll do some water skiing and tubing and stuff like that. So that'll be fun. I don't we'll see if I get up on water skis. It's been a few years, but I do like water skiing, and certainly tubing's real easy, John. That's that's not hard at all. But uh we'll we'll see if I wanna water skiing's a young man's sport, John. I don't know if you've ever done it, but it's for young people.

SPEAKER_02 1:16:02

No, no, I I to be honest, I'm terrified of water. Um and so yeah, you're not gonna get me. I'm quite happy to go in a boat, but anything that takes me outside of the safety of a boat, I'm not no no.

SPEAKER_01 1:16:13

I've it's not gonna happen.

SPEAKER_02 1:16:15

Oh, I love the ocean. That's that's that's I just I'm just boat.

SPEAKER_00 1:16:19

I don't want to be in it.

SPEAKER_02 1:16:21

Deep water freaks me out, so yes.

SPEAKER_00 1:16:23

Yeah, yeah. Well, all right, John. Uh I think that wraps it up. And you know, safe travels to you, and we'll see you in a month.

SPEAKER_02 1:16:30

Yeah, have yeah, have a great 4th of July. I I I I was not trying to be down about it at all. It's just like I say, it's it's one of those things that I I appreciate other people's traditions. It's just we have we have different traditions, different upbringings, you know, we look at things different ways, but I hope you have a blast.

SPEAKER_00 1:16:47

I don't know how we've gone this long without mentioning the World Cup, which is incredibly exciting here in Kansas City. We you know, we got our last game tomorrow night, and then there's a good chance that if Portugal and Argentina match up in one of the knockout rounds here, that that that'd be here in Kansas City. So that's got us a buzz. And but it's been the World Cup's been pretty, pretty enjoyable. I think it's a much better record for attending.

SPEAKER_02 1:17:07

We're sad to see Scotland on the brink of going out because I don't think they're gonna make it.

SPEAKER_00 1:17:12

And they had a tough draw, John.

SPEAKER_02 1:17:13

But absolutely that group that group was horrible, and so I'll sort of switch allegiance to England.

SPEAKER_00 1:17:19

They had a nice win, you know, they've drank they they've absolutely I don't know if you know the story about when they came over to they flew over into Boston, they they wiped the plane out of beer, they're wiping the Boston bars out of beer, they're wiping the Miami bars out of beer, you Scots are taking it serious, man. It's funny.

SPEAKER_02 1:17:35

I just I just look on and say, well, I'll I can choose to be a Scot occasionally, but ultimately I'm English. But when it comes to the world World Cup, because when we when they qualified, we were back in Scotland last November and everyone was so excited about qualifying. So even even Kath, my wife, um she she has isn't interested in football at all, but she's been insisting on watching the the football games or the Scottish matches because because she she was there and and got sort of whipped up in the the um Scottish fervor at qualifications. So yes. I was like I said, I'll switch to England because that's who I more typically follow. They struggled in their last game, though.

SPEAKER_00 1:18:21

They have a they have a potential to go far. If you really want to see something funny, go look at the 20,000 Dutch that were doing the little dance in the streets of Kansas City. I don't know if you saw that. There were 20,000 of them doing something. Moving back and forth and stuff, and it was wild. It's got a nice buzz here. Kansas City's really penetration. But okay, John, take care. Have a great vacation.