xMAP® Connect

Welcome and Introduction to Luminex

Angelo Rago
President, Luminex

Originally presented October 2023 at xMAP^® Connect in Shanghai, China

Luminex kicks off the 2023 xMAP^® Connect event in Shanghai, China and provides an insightful overview of Luminex and xMAP^® Technology. Discover how xMAP Technology is transforming the landscape of multiplexing and driving innovation in various fields.

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xMAP^® Technology: The Year in Review

Sherry Dunbar, PhD, MBA
Senior Director, Global Scientific Affairs Programs, Luminex

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

xMAP^® Technology has been the gold standard multiplexing platform for biological research and discovery for 28 years. Beginning as a novel suspension array multiplexing technology, it has become the worldwide leader for bead-based multiplexed analysis and an important tool in the toolbox for research and clinical laboratories around the world. As of September 2023, more than 70,000 peer-reviewed publications have been reported by investigators who use xMAP Technology in their work. This session will review some recent xMAP literature that highlight the versatility and flexibility of this innovative platform. We will discuss a variety of topics that all have one thing in common – xMAP Technology. Join us as we discuss some of the newest publications on biomarker research in cancer research, cytokines, serology, and immunology.

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Illuminating the Host Responses to Visible Light in the Respiratory Tract Via Use of Thermo Fisher Assays on the Luminex Platform

Jacob F. Kocher, PhD
Director of Virology, EmitBio

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

Biotech startup EmitBio is developing a handheld device for at-home treatment of respiratory infections using specific light wavelengths to eliminate pathogens. In this presentation, EmitBio’s Director of Virology, Jacob Kocher explains how the company employed xMAP^® assays to study cytokine responses, using the data to prioritize or rule out certain wavelengths. Notably, they discovered that 425 nm light is broadly effective against various SARS-CoV-2 variants.

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Evaluating Neuroinflammation During COVID-19 and Other Neurotropic Viral Diseases Using MILLIPLEX^® Kits

Mukesh Kumar, PhD
Associate Professor Georgia State University

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

Increased production of cytokines and chemokines in the central nervous system (CNS) is a common event observed during infection with neurotropic viruses. These virus-induced cytokines and chemokines play an important role in the modulation of host defense and disease manifestation. Mukesh Kumar, from Georgia State University, routinely uses MILLIPLEX^® Mouse Cytokine/Chemokine Magnetic Bead Panel to examine the inflammatory response in neurotropic flavivirus infection in mice and its association with disease severity. They have used various knockout mouse models to directly address the role of various inflammatory and inflammasome molecules in flavivirus neuropathogenesis. Interestingly, we successfully assessed the protein levels of multiple cytokines, chemokines and growth factors in Zika virus-infected guinea pigs using a MILLIPLEX^® Rat Cytokine/Chemokine Magnetic Bead Panel. Their data demonstrate that ZIKV infection resulted in a significant increase in protein levels of IL-2, IL-5, IL-18, IL-12 (p70), TNF-α, G-CSF, MCP-1, MIP-1α, LIX, Fractalkine and VEGF in guinea pigs. My laboratory was one of the first to establish a K18-hACE2 mouse model to study COVID-19 disease. Using this model, they demonstrated that SARS-CoV-2 attacks the brain, not lungs, which triggers severe disease in this mouse model. Using a MILLIPLEX^® Mouse Cytokine/Chemokine Magnetic Bead Panel, we detected significant production of cytokines/chemokines in the CNS of K18-hACE2 mice. In addition, Georgia State University and others have used the Luminex platform and Milliplex kits to show that infection of mice with SARS-CoV-2 variants result in distinct tissue-specific proinflammatory cytokine signatures. Together, these data provide insights into the pathogenesis of a neurotropic virus infection in mice.

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Antibody Status Made Simple An Innovative Application of Luminex xMAP^® Technology

William F. Shea
Chief Executive Officer ImmunoProfile

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

ImmunoProfile^® has built the first ever serology test panel to simultaneously measure IgG antibody levels across multiple vaccine-preventable infectious diseases to determine protection levels (titer tests). Specimens are collected by finger-prick at any location convenient for the user. Standardized test results are provided electronically via a single page PDF that can be easily presented, uploaded, forwarded, or stored in a private portal making credentialing faster, more convenient, and significantly less expensive.

Diseases reported on the panel are Diphtheria, Pertussis, Tetanus, H. Influenza B, Hepatitis A, Hepatitis B, Measles, Mumps, Rubella, Polio, Varicella Zoster/Chicken Pox.

The test for each disease is built to the World Health Organization (WHO) standards for antibody levels that are shown to provide protection to that disease. The test panel has been validated and approved for use in all 50 States and Washington DC. Comparative test results are shown on the ImmunoProfile website at www.immunoprofile.com.

The ImmunoProfile proprietary assay runs on all Luminex Platforms including the MAGPIX^®, LUMINEX^® 100/200^™, and FLEXMAP 3D^®. The specimen is collected on a Dry Blood Spot card from a finger-prick. Comparison studies between Serum and a Dry Blood Spot have been performed and showed equivalency. An FDA Usability Study was passed on the specimen collection kit and workflow process of a user collecting their own specimen, registering on the ImmunoProfile website, and sending the specimen to the lab using the preaddressed UPS shipping pouch.

This multiple infectious disease panel is ideal for individuals wanting to know their protection levels to infectious disease (for multiple reasons; including travel, preconception planning, visiting grandchildren, etc..), for individuals needing to present proof of their status to third parties (medical professionals, immigration seekers, occupational health, etc..), and for surveillance by public health authorities to learn the prevalence of protection levels to dangerous diseases in any geographical region or community.

Whatever the reason, ImmunoProfile makes knowing an individual’s protection level, or a herd’s protection level, to these infectious diseases easy and inexpensive to obtain. Anyone can purchase the entire 11-disease panel. The specimen is collected via a finger -prick onto dry blood spot card at any location avoiding the blood draw visit.

For more information about this test panel please visit ImmunoProfile at www.immunoprofile.com.

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Optimizing Cytokine Analysis: Choosing the Right Assay for Accurate Detection and Interpretation of NHP Cytokine Expression

Laura L. Holcum Stone, PhD
Senior Research Associate University of Minnesota

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

Cytokine profiling is a valuable tool used in preclinical research for monitoring an immune response and cell signaling pathways during disease progression and treatment. The increasing availability of commercial multiplexed cytokine assays has made measurement of circulating cytokine levels more common in the study of the pathogenesis of disease, but interpretation can be complex, often due to the lack of assay validation and consideration of how unintended model-imposed variables may affect expression. Recognizing the burden of chronic illnesses and evolving infectious disease threats, the ability to accurately interpret preclinical data in disease and therapeutic safety and efficacy studies is more important than ever. Determination of the ‘fitness’ of an assay for a given application as well as a thorough understanding of how elements of experimental design may impact study outcomes are critical to the generation of robust and rigorous research data.

Nonhuman primates (NHPs) are often chosen as a preclinical model of disease and treatment due to their close evolutionary relationship with humans, including a complex immune system. These studies are often the critical last step before translation to clinical application. Similar to the clinical situation, it is critical to understand how demographic characteristics and experimental design, such as species and sex, may effect therapeutic efficacy or safety. Likewise, experimental factors that might have no clinical relevance, sedation or restraint, can also impact study outcomes in ways that may confound results and hinder clinical translation. Here we will outline the importance of qualifying commercial cytokine assays prior to use in preclinical research, as well as demonstrate how common study-imposed variables may impact the expression and detection of cytokines.

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Development of a Pharmacodynamics Assay for TOS-358, the First Covalent Inhibitor of PI3Kα in Clinical Development, with LuminexPLORE Lab

Raymond Mak, PhD
Scientist, Totus Medicines

Originally presented November 2023 at xMAP Connect^® in Atlanta, GA

At Totus Medicines, xMAP^® Technology is being used to support a pharmacodynamics assay to validate the clinical performance of a molecule designed to inhibit PI3Kα, one of the most mutated oncogenes. Raymond Mak described working with the LuminexPLORE Lab to convert assays originally developed with western blot technology to a dual-reporter xMAP assay for the xMAP INTELLIFLEX^® DR-SE System that would measure the percentage of PI3Kα that bound to the candidate drug.

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Immunoassay Design and Development for Early Detection of Colorectal Cancer on the xMAP^® Platform

Jason Liggett, PhD
Chief Scientific Officer New Day Diagnostics (formerly EDP Biotech)

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

A look at the principles and process of immunoassay design, the significance of biomarkers selection, the optimization of assay parameters for enhanced sensitivity and specificity, and strategies to overcome challenges in assay development.

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How the best multiplexing technology got even better – xMAP INTELLIFLEX^®

Sherry Dunbar, PhD, MBA
Senior Director, Global Scientific Affairs Luminex

Originally presented November 2023 at xMAP Connect in Amsterdam

Fall in love with xMAP^® Technology all over again. Over the past 28 years, xMAP Technology has become the worldwide leader in bead-based multiplexing. The new xMAP^® INTELLIFLEX lever- ages all of the benefits of xMAP to deliver capabilities that will allow users to expand their research even further. INTELLIFLEX is the only compact, flow-based multiplex platform that combines the proven performance of xMAP Technology with modern features to empower assay developers with enhanced performance and an improved overall user experience. No other multiplex platform combines low- and high-plex capabilities, quick time to reliable results, and the ability to acquire data for two parameters per analyte simultaneously. In this presentation, you’ll learn all about the INTELLIFLEX system’s hardware and software features. We will discuss some of the exciting new applications made possible by the xMAP INTELLIFLEX System and review key studies that focus on recent work in dual reporter applications in serology and cancer research.

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Measuring the humoral response to SARS-CoV-2 using plasma and fingerstick blood

Dylan George
Graduate Research Assistant, University of Nebraska

Originally presented November 2023 at xMAP^® Connect in Atlanta, GA

Background
Understanding the role of antibody-mediated immunity at the population level is valuable to inform public health decisions relating to SARS-CoV-2. While many immunoassays currently exist that can each provide their own unique insights into the humoral response, many of these are complicated, time-consuming, and require specific specimens. Additionally, many of these off-the-shelf ELISAs and LFAs only give results relating to a single antibody target.

Methods
Using the xMAP INTELLIFLEX^®, they developed a multiplex immunoassay (HumorX) that simultaneously measures antibodies targeting the SARS-CoV-2 nucleoprotein (N) as well as multiple variants of the receptor binding domain (RBD) including those from canonical and Omicron BA.5 strains. Recombinant proteins were coupled in house to MagPlex^® Microspheres. Assay performance was characterized using plasma samples collected before and during the COVID-19 pandemic. Samples represent multiple study cohorts. This testbed has been developed for use in human serum and plasma, and while this has enabled great results from subjects admitted to the hospital, it leaves a gap of individuals who may not be presenting for care. To enable community screening and large scale sample collection, we adopted the MITRA fingerstick device (Neoteryx) that allows volumetric sampling of fingerstick capillary blood. Processing of fingerstick samples was optimized to correlate antibody levels with plasma.

Results
HumorX demonstrated high clinical specificity (99.3% anti-RBD/N IgG, 99.6% anti-RBD/N IgM) and high clinical sensitivity (100% positivity 15-90 days post-PCR confirmed COVID-19). HumorX shows an excellent correlation between plasma/ serum and fingerstick collected blood for anti-RBD (R2=0.9191) and anti-N (R2=0.9414) antibodies.

Conclusions
This test has been made possible thanks to the use of xMAP^® Technology combined with in-lab developed protocols. Fingerstick blood is a promising sample matrix that is less invasive to collect and can be done at home or widely applied at community events.

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