New Year, New Cookbook: Check Out This Great Resource for xMAP® Technology Users

The fifth edition of the xMAP Cookbook offers new protocols and application examples, with additional tips, tricks, and insight from our experts

We’re delighted to announce the xMAP Technology has been a game-changer for scientists around the world, but for those who are unfamiliar with it, we try to provide a primer on multiplexing and how it can benefit scientists across research fields. As a bead-based multiplexing tool, it can be used for both proteomic and genomic applications, and xMAP in particular helps reduce sample and reagent volumes while generating more information.

With xMAP, you can analyze up to 500 targets in a single run

For technology that’s so different from standard, single-analyte immunoassays, we’ve found that our customers really like to have a guide showing them the many different ways it can be used. That’s why we created the original xMAP Cookbook back in 2013. Since then, we’ve regularly updated this handy collection of protocols to include updated tips and tricks based on what we see in our labs, but also what our customers are doing in the field.

All xMAP Cookbooks include detailed instructions for designing, creating, optimizing, and validating your xMAP assays, plus assay development guides for various applications, a detailed list of common xMAP buffers, and a comprehensive list of equipment needed to run xMAP assays.

In our just-released fifth edition, we’ve added some exciting new material—plus tips and protocols for our newest xMAP platform. Here’s a quick look at some of the highlights:

Webinar: Designing a COVID-19 Vaccine Efficacy Assay with the xMAP INTELLIFLEX® System

In Chicago, scientists employed the dual reporter channel to generate more data without incurring more costs

We always enjoy seeing ® Systems” href=”/intelliflex/#overview”>xMAP INTELLIFLEX® System.

If you don’t have time for the ® System” href=”/flexmap-3d-system/#overview”>FLEXMAP 3D® System, the automated workflow permitted testing more than 1,000 samples per day. Dr. Borgia and his team validated the test on both plasma and dried blood spot samples.

Validation studies in more than 1,000 samples showed that the assay featured “near-perfect sensitivity and specificity,” Dr. Borgia said. “Our overall accuracy was over 99 percent.”

Vaccine efficacy testing with xMAP INTELLIFLEX

Later, the team created a separate serology assay to quantify the immune response to COVID-19 vaccination. By this time, the lab had adopted a beta version of the xMAP INTELLIFLEX® System, which provides enhanced multiplexing capabilities with the addition of a dual reporter channel. This feature makes it possible to evaluate two isotypes at a time—such as IgG and IgM—and generate twice as much data from the same sample and reagent volumes as any other xMAP assay.

The new assay focused on antibodies to the spike, nucleocapsid, and matrix proteins, analyzing IgG, IgM, and IgA for each. Dr. Borgia described a number of validation and optimization steps for the new instrument, allowing the team to identify the appropriate incubation time and evaluate intra-assay and inter-assay precision.

What they were really interested in, of course, were the results from the longitudinal monitoring of immune responses to vaccination, particularly among immunocompromised patients. The data they generated supports the recommendation for booster shots, and showed that a third dose helped trigger as much protection in immunocompromised individuals as two doses did in people with a normal immune system.

The xMAP INTELLIFLEX System allowed the team to generate high-quality results at higher throughput and lower cost, Dr. Borgia also noted.

As xMAP INTELLIFLEX systems ramp up next year and beyond, we’re really looking forward to seeing all of the innovative ways the xMAP community put the new dual reporter feature to good use.

For more information about this study, check out the webinar!

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2021 Year in Review: On Why We Do What We Do

After another challenging year, we’re grateful that the work we do here helps

In more precedented times, 2021 might’ve been described as a banner year for Luminex. Despite the persistent challenges presented by the ongoing pandemic, our teams delivered critical assays and reagents, scaled production of our COVID-19 offerings, expanded our team, services, and facilities, and supported scientists and healthcare providers across the world. We did this and more—in the field, from our international offices, from our homes—all while transitioning into a new, integrated company with DiaSorin following our acquisition back in July.

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While there is much to celebrate, we’re aware that against the backdrop of the rising toll of this outbreak—now, day 590 since the World Health Organization declared COVID-19 a global public health threat,1 with more than 276 million confirmed cases and 5.4 million deaths worldwide2 plus new variants to be cautious of—the close to a successful year for our business is hardly a silver lining for what we’ve lost to this pandemic. The last two years have flown by so quickly, most of us haven’t had time to grieve our losses. We’ve adapted to life at a new pace, and there hasn’t been much time to reminisce over our pre-pandemic lives.

There is still hope, however: there have also been 8.5 billion vaccine doses administered across the globe, with much of Europe, China, Australia, New Zealand, Chile, and Canada leading the way with over a 70% vaccination rate.3 Testing and treatment options have also come a long way in just two years—seeing the healthcare community and scientists from all over the world develop solutions at this pace has been incredibly humbling.

As a biotech company, most of us are just grateful that the work we do here helps. It helps improve diagnostics (which helps improve health outcomes), it helps support researchers investigating disease mechanics or developing therapeutics, it helps reduce bench time for busy lab techs. This is why we do what we do.

Forging a path forward

As COVID-19 persevered, our teams worked to increase production of the ® Monitoring Microspheres” href=”/blog/magplex-monitoring-microspheres-let-xmap-users-take-troubleshooting-into-their-own-hands/”>MagPlex® Monitoring Microspheres to our catalog, making good on our promise to continue to deliver tools that can help our customers take their research to the next level.

We also launched LuminexPLORE Lab supports assay optimization, conversion, validation options, and more. With their combined knowledge, no project is too big or too small for our team of experts.

Notably, as we look ahead to 2022, the LuminexPLORE team will move into a new and improved lab space in our Austin headquarters, enabling them to expand their capabilities and throughput. These services can provide a lot of time and resource-saving benefits, so we’re looking forward to hearing what the LuminexPLORE team and the scientists we collaborate with accomplish in the coming year.

A unique combination of specialists

DiaSorin’s acquisition of our company in July marked another big milestone in our 26th year. As experts in immuno- and molecular diagnostics, the merge with a larger, multi-national company will bring the best of both businesses to our customer bases. We’re fortunate to have built a strong community over the years, and with our combined expertise, we look forward to developing additional tools and solutions across the academic, biopharmaceutical, and clinical spaces. How we can help just got better.

Tackling COVID-19 and beyond

We may not know exactly what next year has in store, but as we shared at the close of our first year of the pandemic—we’re dedicated to supporting our customers and partners as long as this lasts. Whether it’s SARS-CoV-2 testing solutions, comprehensive diagnostic panels, or research systems, reagents, and tools, we’re here for you however you need it. We’ll be here when this is over, too.

Warm wishes from our families to yours. Happy holidays from Luminex!

References:

  1. Opening Remarks at the media briefing on COVID-19, 11 March 2020. Dr. Tedros Adhanom Ghebreyesus, WHO Director-General. WHO (Internet). Accessed 2021 Dec. Available from: https://www.who.int/director-general/speeches/detail/who-director-general-s-opening-remarks-at-the-media-briefing-on-covid-19—11-march-2020.
  2. COVID-19 Dashboard. The Center for Systems Science and Engineering at Johns Hopkins University (Internet). Accessed 2021 Dec. Available from: https://coronavirus.jhu.edu/map.html.
  3. Vaccination progress across the world. The Center for Systems Science and Engineering at Johns Hopkins University (Internet). Accessed 2021 Dec. Available from: https://coronavirus.jhu.edu/vaccines/international.

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How Multiplexing Can Support Vaccine Development

xMAP® Technology supported the development of HPV vaccines, continues to provide key results in after-market studies

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Vaccine development is traditionally a complex and lengthy process. It requires a clear and in-depth understanding of the variants of a given disease-causing organism and their modes of pathogenesis, as well as the antigens they express. Successful efforts require a high-quality assay to detect all relevant variants in a timely manner in order to understand immunogenicity, among other critical factors.

It’s no secret that conventional enzyme-linked immunosorbent assays (ELISAs) are time and labor-intensive. For vaccine development, ELISAs simply cannot provide the throughput needed. Thankfully, xMAP® bead-based multiplexing technology offers a flexible and customizable platform to support both gene and protein expression assays. And unlike traditional technologies that can only measure one or a few biomarkers, researchers have the ability to easily scale the number of biomarkers measured—from 1 to 500 analytes—and the ability to customize assays for genotyping, protein expression profiling, gene expression profiling, and more.

Supporting vaccine development when and how it’s needed

A noteworthy example of how scientists have used xMAP Technology in vaccine development comes from the development of human papilloma virus (HPV) vaccines. More than a dozen HPV genotypes are associated with cervical cancer, necessitating a highly robust multiplex assay for the vaccine development process.

Two of the three globally licensed HPV vaccines—Gardasil®, a quadrivalent vaccine (4vHPV), and ® Technology (2021)” href=”/?wpdmdl=49089″>The Development of HPV Vaccines and the Contributions of Luminex xMAP Technology (2021)

  • The Role of Multiplexed Assays in Vaccine Development (2016)

    • Although the vaccines have long since been approved, xMAP multiplex assays continue to power studies examining long-term vaccine safety and efficacy, along with the worldwide impact of vaccination against HPV-related diseases. To learn more about how xMAP Technology can be used for vaccine development and more, check out the xMAP Cookbook!

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    RD-Biotech Expands Antibody and Immunoassay Services with xMAP® Technology

    Joining Luminex’s Licensed Technologies Group enhanced the company’s capabilities

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    Our Licensed Technologies Group represents a who’s who of companies delivering impressive results by applying Luminex’s xMAP® Technology to a wide variety of experiments. One of our newest partners is RD-Biotech, a service provider based in France. The talented RD-Biotech team helps clients with services in immunology, molecular biology, and cellular engineering.

    Small team, big impact

    Founded in 2002, the company works primarily with pharmaceutical and biotech companies interested in outsourcing pre-clinical or clinical tasks for their research and development programs. While the team is small—with a staff of 40—its impact is sizable: RD-Biotech has 200 clients on their roster.

    “We are very active with pharmaceutical and biotech companies,” says CEO Philippe Dulieu. “We do what they cannot do internally.” Common services RD-Biotech handles include antibody development, engineering, and manufacturing; conjugating antibodies to beads; and immunoassay development.

    New ventures

    Recently, Dulieu and his colleagues got a request from a pharmaceutical client working on vaccine development: could they conjugate antibodies to beads for use with xMAP analyzers? The team got started with small batches of beads and quickly ramped up as the project expanded.

    Now, RD-Biotech works directly with Luminex as a Licensed Technologies Partner, which Dulieu says gives his team better access to reagents and customer support. They’re already looking to expand their service offerings around xMAP Technology, and purchased a visit our website.

    Learn more about the benefits of becoming a Luminex Partner here.

    For Research Use Only. Not for use in diagnostic procedures.

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    Getting Double the Data from Less Sample: An xMAP INTELLIFLEX® Dual Reporter Assay in Action

    Using the new INTELLIFLEX System’s second reporting channel, our scientists teamed up with experts at the University of Rochester to design, optimize, and validate their SARS-CoV-2 antibody assay

    If you’ve been eager to see how the new dual reporter channel works on the xMAP INTELLIFLEX® DR-SE System, you’re in luck! In a recent JoVE video, we walked through this process using a multiplex assay designed to detect multiple antibodies to SARS-CoV-2. Here are some of the highlights!

    The assay was designed using xMAP® Technology and is based on work from a team of scientists at the University of Rochester Medical Center. To ensure robust results, they validated the assay with thousands of samples, making it possible to reliably detect the levels of two immunoglobulins in a single reaction while measuring the generation of potential SARS-CoV-2 neutralizing antibodies.

    To help the team take the next step, Senior Field Applications Specialist Dr. Stephen Angeloni and other Luminex scientists worked with them to adapt the assay to the latest xMAP platform—the INTELLIFLEX DR-SE System. This new system offers a dual reporter channel that allows users to generate twice as much data from analytes in the sample, reducing the sample volume and time required to produce the information needed. This unique capability made it possible for the scientists to adapt the assay to detect IgM antibodies in addition to the original IgG responses in a single reaction. The video also demonstrates, for the first time ever, how the assay can be easily modified to measure the generation of potential neutralization capabilities of these immunoglobulins.

    “To our knowledge, this is the first example of a bead-based multiplexing protocol that uses two reporter signals to simultaneously measure two results per analyte,” says Dr. Sherry Dunbar, Luminex’s Senior Director of Global Scientific Affairs, in the introduction to the video. “While this dual reporter method is specific for antibody isotyping, it could be adapted to measure other analyte pairs, such as post-translational modifications or free versus bound drug forms.”

    The rest of the video follows Dr. Angeloni, who demonstrates the dual-reporter assay and key elements of the process, such as optimization. It includes clear, step-by-step instructions to help current xMAP users follow along throughout the protocol.

    The results of Dr. Angeloni & his teams’ efforts are reported in the JoVE video presentation: “A Rapid, Multiplex Dual Reporter IgG and IgM SARS-CoV-2 Neutralization Assay for a Multiplexed Bead-Based Flow Analysis System.” To learn more, check it out on JoVE!

    Watch the video here.

    For Research Use Only. Not for use in diagnostic procedures.

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    AMLI 2021: xMAP® Technology Posters Cover COVID-19 and Biotinylated Beads

    Check out highlights from this year’s conference posters using bead-based multiplexing technology

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    This year, the Luminex team was honored to attend the annual meeting of the Association of Medical Laboratory Immunologists (AMLI) which was held in our hometown of Austin, Texas. In addition to catching up on the latest findings about immunity as well as immune and autoimmune disorders, we presented several posters, and we’re excited to share the summaries with our readers:

    Conversion of a SARS-CoV-2 IgG Titer and Neutralization Assay to Detect IgG and IgM Titers and Neutralization in a Single Reaction

    We teamed up with scientists at the University of Rochester Medical Center to redesign a multiplex assay for SARS-CoV-2 antibodies for use on the new Download the Poster

    Using xMAP Technology to Rapidly Develop and Iterate Antibody Assays for Viral SARS-CoV-2 Proteins: A Feasibility Study

    This poster reports efforts from scientists at the University of Nebraska Medical Center along with Luminex’s Heather Darby, who worked together to rapidly design assays for the COVID-19 pandemic. “As new challenges continue to emerge during the global response to the COVID-19 pandemic, testing that can rapidly deliver tailored and relevant results will be critical to outbreak surveillance,” they report in the poster.

    The team used xMAP Technology to develop multiplex assays for detecting SARS-CoV-2 antibodies. In this feasibility study, they evaluated a number of variables—such as protein coupling concentration and serum sample dilution factor—and developed and validated the assay in just one week.

    “This assay is ready for further characterization and validation for implementation as a research tool, enabling serological surveillance at a population scale for exposure to SARS-CoV-2, as well as its variants,” the scientists conclude.

    Download the Poster

    Effect of Streptavidin-R-Phycoerythrin Conjugate Size on Fluorescence Signal in Biotinylated Bead Assays

    Scientists from Luminex and Agilent Technologies reported on this project, which aimed to assess the signal given off by different sizes of streptavidin-r-phycoerythrin (SA-RPE) conjugates. “Size differences in SA-RPE conjugates have the potential to affect the fluorescence signal obtained in different assay setups,” they note.

    They found that the use of a longer linker with biotin-coated beads leads to stronger signals with SA-RPE, regardless of size. In addition, larger conjugates of SA-RPE produced more signal in certain situations.

    “Overall, the optimal SA-RPE conjugate for a given assay depends on the assay chemistry,” the team writes. “Users should test different SA-RPEs to determine what works best for their assay.”

    Download the Poster

    For the latest developments from our colleagues and collaborators + all things xMAP, check out our blog series!

    For Research Use Only. Not for use in diagnostic procedures.

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    Developing a New Multiplex Assay for Early Detection of Colorectal Cancer

    Luminex Licensed Technologies Partner Spotlight: EDP Biotech

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    Early detection makes a real difference in outcomes for cancer patients. But for cases of colorectal cancer, detecting tumors in the earliest stages has been virtually impossible. The team at EDP Biotech is committed to changing that by developing a simple, accurate, blood-based diagnostic test for biomarkers of colorectal cancer. They’ve chosen to build this assay on colorectal cancer. We are working on our first multiplex assay to aid in early detection. Most of the other testing options available today are expensive, highly invasive, or simply messy—patients don’t want to take the test. We want to build on our experience in colorectal cancer to offer a simple blood test. For our staff, this is a personal mission.

    How would early detection make a difference?

    If colon cancer is caught early, the patient outcome is drastically improved. Five-year survival rates just plummet as you move from early stage to later stages. Most of these cancers are caught in late stage, when five-year survival is less than 15%. Most patients aren’t getting screened, even when they have increased risk.

    How are you using xMAP® Technology?

    The xMAP platform offered a really attractive option for screening a large number of biomarkers quickly while consuming a small amount of serum. We started with a large number of biomarkers and then narrowed those down to a dozen or so that we’re interested in putting into a single kit. We worked with MilliporeSigma for the custom development and manufacturing reagents for the single kit. We started working with the ®” href=”/flexmap-3d-system/”>FLEXMAP 3D® and paired it with an automated liquid handler for higher throughput. The technology has allowed us to grow and expand as our knowledge increased.

    What will the EDP colorectal test look like when it’s ready?

    We are hoping to launch the test next year. The final product will be a combination of the xMAP-based assay with our proprietary machine learning algorithm which takes those quantified values and converts them into a yes/no answer to help physicians determine whether they should recommend a follow-up colonoscopy to their patients.

    How has your experience as a Licensed Technologies Partner been?

    As a small company looking for a very robust and high-throughput technology that’s very sensitive, working with Luminex has been a good fit. There are also a lot of other Licensed Technologies Partners out there willing to help us. It feels like being part of a really tight-knit community of experts.

    What’s next for the EDP team?

    We are a small diagnostics company, and during the pandemic we responded quickly to help our community with COVID testing. As the world begins to return to its “new normal,” we will return to all hands on deck for the colorectal cancer assay. Based on the success of this test, though, we do have some other tests in the pipeline for early detection of other solid-tumor cancers and infectious diseases.

    You can view a full list of our partners by visiting our website, or if you’re interested in becoming a partner, you can learn more here.

    Learn more about the benefits of becoming a Luminex Partner.

    For Research Use Only. Not for use in diagnostic procedures.

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    MagPlex® Monitoring Microspheres Let xMAP® Users Take Troubleshooting into Their Own Hands

    Monitoring beads help users troubleshoot unexpected median fluorescent intensity patterns and weak signals, providing added confidence in your assay results

    We’re excited to announce the availability of our new “It’s very handy to have these beads in the reaction so if a problem does arise, you can quickly identify whether it’s on the instrument or the assay side,” says Stephen Angeloni, PhD, Senior Field Application Scientist at Luminex. “They’re useful for quality control in an assay.”

    Before these beads became a commercially available product, Dr. Angeloni used them to help customers who were having technical challenges with their assays, such as weak signals or unexpected fluorescence patterns. Adding monitoring beads made it easier to troubleshoot the cause of those issues.

    Since using the monitoring beads, Dr. Angeloni has helped customers identify some common culprits of assay issues, including degraded reagents, forgotten detection reagents, and reagents that were pipetted in the wrong amount. When these types of issues arise, being able to pinpoint which one is the cause behind an assay that just isn’t working optimally can save researchers a lot of time.

    A glimpse into the beads

    The beads are magnetic polystyrene microspheres that have been color-coded to classify to a specific region while providing reporter MFI values specific to instrument type and reporter modes. The reporter dyes are inside the beads rather than on the surface, so there is no modification necessary to use them in the regular xMAP workflow. They can simply be combined with the other beads needed for an assay.

    MagPlex Monitoring Microspheres require the use of xPONENT® or INTELLIFLEX software, and two products are available depending on the reporter channel being monitored:

    We’re constantly impressed by the different ways our customers use our products to meet their unique needs, and we’re committed to developing solutions that optimize the xMAP Technology experience, like these new microspheres. To learn more about them, take a look at our ® Monitoring Microsphere Sales Sheet” href=”/?wpdmdl=49087″>download a flyer with more information.

    Learn more about MagPlex Monitoring Microsphere here.

    For Research Use Only. Not for use in diagnostic procedures.

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    Luminex Licensed Technologies Partner Spotlight: Wageningen Prime Diagnostics

    Multiplex molecular testing for plant pathogens

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    In the agricultural community, there’s a need for high-quality molecular testing. Pathogens can appear without warning and decimate entire crops. Farmers, plant breeders, and other members of the agricultural community need access to simple, affordable, and accurate tests for known and emerging threats to monitor plant pathogens when they first appear and prevent catastrophic crop or resource loss.

    Wageningen Prime Diagnostics was founded two decades ago to address this need. As part of the Wageningen University & Research in the Netherlands, Prime Diagnostics develops and sells a large portfolio of test kits for crops, garden flowers, and other economically important plants.

    Scientists at Wageningen Prime Diagnostics often design multiplex assays to allow customers to test for many pathogens at once. They use Luminex’s bead-based multiplexing disease in potato plants.

    Implementing xMAP Technology was important for achieving the level of multiplexing needed for plant pathogen tests. “Some garden plants are notorious for having a lot of pathogens attack them,” van Beckhoven says. “In plant propagation, they multiplex a lot because they have to focus on so many different pathogens. Luminex is perfect for that.” Recently developed tests include an 18-plex kit for petunias, as well as a group of tobamoviruses that infect tomatoes, peppers, potatoes, and eggplants.

    “Our experience with Luminex has been very good,” says Bergervoet, who develops many of their xMAP-based tests. “The technology is very robust and the bead quality is very consistent.”

    That consistency is especially important because customers often run high-volume operations, testing thousands of samples each day in fully automated pipelines. “We looked at what our customers wanted and what Luminex could offer, and it was a good match,” van Beckhoven says. “We have different platforms we develop tests on, and Luminex is the easiest and most robust. It’s almost foolproof.”

    Wageningen Prime Diagnostics is one of our exclusive partners that span a wide range of industries and specialties. You can view a full list of our partners by visiting our website, or if you’re interested in becoming a partner, you can learn more here.

    Learn more about the benefits of becoming a Luminex Partner here.

    For Research Use Only. Not for use in diagnostic procedures.

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