xMAP® INSIGHTS Plant and Animal Issue

xMAP INSIGHTS ®

Plant and Animal Issue

Dutch Scientists Subtype Antibodies Against Avian Influenza Virus Subtypes in Poultry Sera with xMAP® Technology

Brazilian Scientists Produce Antitoxin and Antivenom Treatments with xMAP® Technology

Read more on page 4

Pictured: Dr. Milena Akamatsu

Pictured: Sandra Venema

Read more on page 6

xMAP INSIGHTS ®

A Magazine for the Multiplexing Community Hello, My name is Chris Haake, and I am pleased to be the guest editor of this edition of the xMAP® Insights Magazine. This publication started in 2016 and continues to connect scientists in the multiplexing community by sharing innovative applications of xMAP® Technology while providing valuable resources to catalyze your next experiment.

This issue focuses on the use of multispecies xMAP assays for the medical and agriculture industries. Here are just a few of the stories shared in this issue: • "Scientists in Brazil Produce Antitoxin and Antivenom Treatments." They have chosen Luminex technology to monitor the immune response in horses before collecting serum to make these lifesaving treatments.

• "Scientists in the Netherlands Subtype Antibodies Against Avian Influenza Viruses in Poultry Sera with xMAP Technology."

• "Luminex Partners Bring the Power of Multiplexing to Scientists Around the World."

I hope you enjoy this issue as much as I do. Don't hesitate to contact me directly at chaake@luminexcorp.com with future story ideas you might have.

Warm Regards,

Chris Haake M(ASCP), MBA Senior Strategic Manager, Marketing Global LTG Luminex Corporation A DiaSorin Company

2

CONTENTS 2

Letter from the Editor

4

Dutch Scientists Subtype Antibodies Against Avian Influenza Virus Subtypes in Poultry Sera with xMAP® Technology

6

Brazilian Scientists Produce Antitoxin and Antivenom Treatments with xMAP® Technology

8

xMAP® Houston Highlights

10

Luminex Partners Bring the Power of Multiplexing to Scientists Around the World

12

LuminexPLORE Lab Fast Facts

Dutch Scientists Subtype Antibodies Against Avian Influenza Virus Subtypes in Poultry Sera with xMAP® Technology Chris Haake

Multiplexing allowed them to reduce sample volume, save money, and report results faster Many countries in Europe and North America have reported their worst numbers of avian influenza (bird flu) in recent years. This dangerous virus—which infects food animals such as chickens, as well as dozens of wild bird species, can force farmers to cull their flocks.

One important way to monitor the avian influenza landscape is by detecting antibodies to characterize any relevant virus strains to which a flock has been exposed. Unfortunately, the diversity in influenza strains makes this very challenging. Each virus contains a specific subtype of haemagglutinin (HA) and neuraminidase (NA).

For avian flu, there are 16 known HA subtypes and nine known NA subtypes, which can occur in different combinations. With traditional testing techniques, individual assays must be run for each HA and NA subtype until the correct one is identified. This is a time-consuming, tedious approach.

At Wageningen Bioveterinary Research (WBVR) in the Netherlands, scientists have adopted xMAP® Technology for a multiplex, beadbased approach to avian influenza antibody testing. The assay they created contained all 25 subtypes of HA and NA, which allowed them to generate complete serotyping information from a single reaction.

Plus, multiple samples could be run at the same time for a truly scalable, high-throughput approach.

“Getting results within a day” Moreover, WBVR is responsible for testing the collected samples at farms throughout the year, wherein the lab then looks for the subtypes of the low pathogenic avian influenza viruses circulating throughout the Netherlands. The test, developed by a team of scientists which included veterinarian Evelien Germeraad and lab technician René Achterberg, is essential for avian influenza monitoring in poultry and in other birds. “This assay made it possible to do the subtyping of the antibodies in the poultry sera really quickly,” Germeraad says. “That process usually takes a lot of time and is expensive. Now we can get results within a day.”

Achterberg’s lab team turned to xMAP Technology when the assay they had previously used was discontinued by its manufacturer.

The team was already running Luminex xMAP Technology, so that platform “was the logical successor,” Achterberg says. Developing

the new test was a relatively straightforward process of coupling different variants of the HA and NA proteins to xMAP beads, and then optimizing the assay for cross-reactivity and other characteristics.

The flexibility of xMAP Technology allows for simultaneous sample testing

Evelien Germeraad and René Achterberg

The resulting test was a significant improvement compared to their prior methods, e.g., the hemagglutination-inhibition (HI) assay. The classical HI assay requires hundreds of microliters of serum for HA subtyping, while with xMAP Technology, only two microliters of serum is sufficient to generate comprehensive results. The workflow was reduced from at least two days to half a day. And since the chickens have limited serum, using less volume for the assay allows for repeats, if needed. The new assay also makes it possible to test many more samples at once and can increase throughput as needed.

The flexibility of xMAP Technology is another important advantage for scientists at WBVR. Since the team published their first xMAP-based avian flu assay design in 2019, they have continued to optimize it and have since added some new variants of key proteins. “That’s a great feature of Luminex assays, that you can easily add more proteins if needed,” Achterberg says. “If you use a commercial assay, you have to rely on what they include. We can include whatever we want.”

The assay now includes multiple variants for every HA and NA subtype. “Avian influenza has so many subtypes, so you really need to be able to multiplex in a test,” Germeraad says. “That’s why this assay is such a great solution.”

Learn more about Wageningen Bioveterinary Research here.

xMAP® INSIGHTS | Spring 2022 4

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xTEND YOUR

RESEARCH NOT YOUR WORKDAY

xMAP® Technology helps deliver more data from less sample, giving you some of your time back in an increasingly busy world. Additional benefits include: • A broad dynamic range, helping you avoid repeat testing due to out-of-range values • A compact footprint, with an embedded PC and an easy-to-use touchscreen interface • Intuitive and flexible software • An xTENSIVE menu of commercially available kits • Custom assay support, including assay development services through the LuminexPLORE Lab

Whatever your needs are, xMAP Technology offers ways to save. Learn more about the many benefits of multiplexing with xMAP INTELLIFLEX® on our website: luminexcorp.com/INTELLIFLEX.

For Research Use Only. Not for use in diagnostic procedures. xMAP® INSIGHTS | Spring 2022 © 2022 Luminex Corporation.

A DiaSorin Company.

All Rights reserved.

Luminex , xMAP INTELLIFLEX, and xMAP are Luminex supports life science research with our Research Use Only (RUO) product portfolio. Not for use in diagnostic procedures.

5 trademarks of Luminex Corporation, registered in the US and other countries.

Brazilian Scientists Produce Antitoxin and Antivenom Treatments with xMAP® Technology Chris Haake

They have chosen xMAP® Technology to monitor the immune responses in horses before collecting serum to manufacture treatments

Of course, the process is not nearly as simple as waiting for the animal to produce antibody-rich serum. From the time the horses are immunized with a toxin or pathogen, scientists Milena Akamatsu and Paulo Lee Ho must closely monitor their equine patients’ immune responses. To do so, they have chosen xMAP Technology.

Scientists at Instituto Butantan in Brazil produce a great majority of the antivenom, antitoxin, and vaccine treatments in the country. They manufacture the 140 million flu shots offered annually by Brazil’s Ministry of Health, making the institute the largest producer of influenza vaccines in the southern hemisphere.

While flu vaccines are common around the world, the institute’s other products are more specific to Brazil’s needs, such as antivenom treatments for snakes, scorpions, and spiders. Scientists there also produce antitoxins for diphtheria, tetanus, and botulinum, as well as an anti-rabies immunoglobulin.

Their team has incorporated a multiplex panel of 23 cytokines and chemokines for testing horse plasma collected at the time of immunization and again later as the immune system responds to the new threat. “We look for immunological markers associated with good response and bad response,” Akamatsu says.

“We can analyze several analytes at the same time from a small amount of sample” Each treatment can be adjusted slightly to optimize performance. “We are using new formulations for old antigens. We usually formulate with new adjuvant,” Ho says. “Then we want to know the kind of response the new adjuvant induces in this new formulation to see if the response is associated with better immunogenicity of the antigen.”

Their team is constantly looking for ways to improve both the products they generate, and the methods they use, Akamatsu notes. While there are other techniques they could apply when monitoring the immunological response in horses, “We chose Luminex technology because it’s easier,” she adds. “We can analyze several analytes at the same time from a small amount of sample.” And while small sample volume is not as critical for horses, it makes a difference with mice and other small animals that produce limited serum.

xMAP Technology offers a flexible, scalable solution With xMAP Technology, scientists can query up to 500 protein or nucleic acid targets in a single reaction using very low sample volume.

This low volume can allow flexibility for repeats, if needed. And by allowing users to run multiple samples at once, costs are reduced by minimizing the use of reagents.

Dr. Milena Akamatsu

Transforming serum into treatments through xMAP Technology Many of these treatments are based on serum produced from an animal’s natural immune response to toxins or venoms. At Instituto Butantan in Sao Paulo, scientists often work with horses—which are not only docile and easy to handle, but they’re large enough to produce significant volumes of serum that can be transformed into treatments.

At the institute, these features are essential as scientists seek to create a broad variety of antitoxin and antivenom treatments, in addition to their thriving vaccine production. Each treatment is different, and scientists must have a flexible, scalable solution that can be implemented as needed.

xMAP® INSIGHTS | Spring 2022 6

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Expanding Possibilities in Animal Health

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Verified: All assays are fully verified for use in the species/sample types indicated.

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All Rights Reserved. MilliporeSigma, the vibrant M, Millipore, and MILLIPLEX are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources.

MilliporeSigma is the U.S. and Canada Life Science business of Merck KGaA, Darmstadt, Germany.

xMAP Houston Highlights Caitlin Butterworth This past month, Luminex’s xMAP® Connect event took place in Houston, TX, providing occasion for leading multiplex experts to converge and build upon the collaborative reservoir of knowledge shared amongst this exclusive community of xMAP insiders.

Houston’s xMAP Connect conference offered an elite lineup of speakers specializing in diverse fields across health science, all of whom having furthered their research through xMAP Technology.

Here are some key highlights from their presentations.

in India while 100 million more vaccine doses have been approved and designated for individuals in Botswana.

Dr. Maria Walker, an assistant professor at the University of Houston who works in the University of Houston Eye Clinic, introduced xMAP Connect attendees to her experience implementing xMAP Technology to analyze human tear fluid in order to better understand the biology behind dry eye. Making use of xMAP Technology’s flexibility, Dr. Walker ran a cytokine panel on tear fluid—a notoriously complex substance—to search for interleukins and matrix metalloproteinases.

While her work with dry eye continues, Dr. Walker seeks to use all mechanisms of her research to build a better understanding of dry eye and its most effective treatment options; she also hopes her study will help establish protocol for tear fluid analysis in the future.

One such speaker was Dr. Fraser Symmans, pathologist and serving director at the MD Anderson Cancer Center. Dr. Symmans and his research team established a custom, xMAP-based assay to investigate a group of genes capable of predicting how individuals with HR+/HER2breast cancer will likely respond to endocrine therapy.

Illustrating the flexible application of xMAP Technology further, Dr. Jair Espindola-Netto of the Mayo Clinic expressed how he and his research team leveraged xMAP Technology to better understand cell senescence and its role within aging. In studying anti-aging senolytic drugs, Dr. Espindola-Netto relies upon xMAP Technology to help characterize proteins within cells linked with senescence so he and his team may better understand how susceptible cells will respond to senolytic medication. With the help of xMAP Technology to collect data efficiently, reliably, and precisely, Dr. Espindola-Netto and his team are better able to understand how individuals may best combat the degenerative effects of aging and remain healthier for longer.

Dr. Jackie Surls, Director of LuminexPLORELabs, Former Luminex intern Atharwa Monthe and Dr. Stephen Angeloni

In sharing how xMAP Technology has supported some of the most dynamic points of their research, the participants of xMAP Connect Houston were able to not only acquire a better understanding of xMAP’s flexible application and numerous capabilities, but build upon the knowledge, experiential understanding, and potential this technology empowers in a community environment defined by collaboration and collective progress.

On choosing xMAP Technology to advance his work, Dr. Symmans explained that he and his team “made the choice for (an) xMAPbased (assay) on some preferences and some workflow logistics that (they) thought would be advantageous.” Not only did xMAP Technology offer “outstanding performance;” the xMAP assays’ “strong reproducibility” and “excellent concordance in different labs,” made it a flexible and “very stable platform” for Dr. Symmans’ work.

Also presenting was Dr. Jeroen Pollet, an assistant professor at Baylor College of Medicine and a director on the Vaccine Development Team at Texas Children’s Hospital. Deploying xMAP Technology, Dr. Pollet and his team were able to test the novel, low-cost alternative COVID-19 vaccine he and his team had developed. With time of the essence, xMAP’s multiplex technology enabled Dr. Pollet to efficiently and effectively interrogate study participants’ immune response to his nascent COVID-19 vaccine. xMAP Technology allowed Dr. Pollet and his team to target multiple analytes simultaneously and acquire reliable data more quickly, expediting the time needed to clear the potentially life-saving vaccine for approval. Dr. Pollet’s COVID-19 vaccine, Corbevax, successfully received emergency use authorization and has already been used to vaccinate 10 million children aged 10-12

Dr. Bobby Bhatia, Luminex Business Manager

Missed the Houston event? We have more xMAP Connect events coming your way. Subscribe here for updates.

xMAP® INSIGHTS | Fall 2021 8

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Trademarks and registered trademarks are the property of their respective owners.

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Luminex Partners Bring the Power of Multiplexing to Scientists Around the World Josh Jenkins, Sales Manager, North America

Luminex’s bead-based multiplexing platform, known as xMAP® Technology, is one of the most widely used assay systems in the life sciences and has now been cited in more than 57,000 peer-reviewed research publications. But here’s something you may not know: xMAP Technology wouldn’t be where it is today without our assay development partners.

could look at the same 10 targets (actually, as many as 500 different targets) for each sample, run the same 96-well plate with minimal sample volume, and obtain results in just four hours.

When xMAP Technology was first invented decades ago, Luminex’s founders realized their small startup company simply did not have the resources to develop assays across the broad range of applications the technology made possible. One of xMAP’s most intriguing characteristics is that it’s basically a blank slate. xMAP beads can be coupled to nucleic acids, proteins, antibodies, modified oligos, carbohydrates, small molecules, and more.

So, the company founders made an unusual choice—one that really set xMAP Technology up for success. They decided to license their powerful technology to larger life science companies. These partners could then use xMAP Technology to build assays and transform them into off-the-shelf products.

Next, our scientific team works closely with the new partner to help them develop and/or optimize new assays. We have field-based scientists who can assist onsite, and we also offer custom assay development services through our LuminexPLORE Lab in Austin. We take our partners’ goals very seriously, and we ensure that each assay is high-quality and reproducible when delivering to customers the best results possible.

Luminex’s partners provide a valuable service to the life science community. If you’re interested in learning more about becoming a Luminex partner, please visit: luminexcorp.com/partners.

Today, our company has about 75 partners around the world who develop and sell xMAP-based assays across applications ranging from biomarker discovery and exosome detection to microRNA analysis and pathogen detection, among others. Collectively, they have brought the power of xMAP Technology to scientists around the world, enabling studies that would not have been affordable or feasible any other way.

Proven Technology

Efficiency

Flexibility

Unmatched in level of end-user adoption and validation

1 to 500 analytes

Allows for both

nucleic acid and protein applications

For our partners, adopting xMAP Technology makes a lot of sense, as they get to bypass the effort and cost it would take to try to build their own multiplexing platform. Instead, they are able to deploy a robust, well-validated system that is already widely installed in research labs.

Thus, their development work can focus on assays that are easier to commercialize. xMAP Technology also benefits our partners’ customers, who need only purchase a single instrument to run these immunoassays and molecular assays rather than having to buy assay-specific instruments. xMAP Technology is easy to use and generates multiple data points very quickly. For example, using standard 96-well ELISAs to examine 10 targets would require enormous amounts of sample and take a good two weeks. With xMAP Technology, a scientist

When new partners team up with Luminex, we help them commercialize by using xMAP Technology. We start by understanding each company’s field of use, and we ensure that the goal is viable and feasible with our technology. Each new partner is granted a nonexclusive license to develop and sell their assays or run them as a testing service (they can also choose to sell xMAP instruments).

>4,000

>57,000 scientific publications cite Luminex technology and/or our partners’ products

simultaneously tested from one sample,

in one well

analytes

can be accessed

on one platform

Meets a wide range of multiplexing and throughput needs with

3 instrument options

>65 510(k) clearances

510 (k)

Open architecture systems enables use of commercial kits and internally developed assays

Partner Facts

>75 Luminex Partner Companies

A wide install base - with

>12,500 xMAP® Instruments Sold Worldwide

1 IVD assay

st The using xMAP Technology was FDA cleared in 2001—

by a Partner

xMAP® INSIGHTS | Spring 2022 10

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Luminex supports life science research with our Research Use Only (RUO) product portfolio. Not for use in diagnostic procedures.

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