Custom Assays: ‘If You Dream It, We Can Multiplex It’

This video presentation details assay development services available from MilliporeSigma

[arve url=”https://www.youtube.com/embed/C_RMvyldKTE” /]

With new biomarkers being published every day, fewer and fewer scientists can get exactly what they need from an off-the-shelf immunoassay kit. That’s why MilliporeSigma offers extensive custom xMAP® Technology assay design and development services, which are showcased in this 10-minute video presentation from Mike Godeny, Head of the Milliplex Reagent Business.

Recognizing that researchers have unique needs and widely varied interests in analytes, Godeny described two approaches MilliporeSigma has for delivering custom assays. The simpler method involves reconfiguring existing analytes from as many as three panels into a single, new multiplex assay.

The more intensive service requires the development of new analytes, multiplexing and verification of performance, and careful validation. For this service, Godeny and his team consult with customers about desired markers, sample type, and species before launching into assay development and design.

Importance of Customer Consultation

In the presentation, he offered examples of multiplex assays developed for Bayer to replace gold-standard ELISA tests, walking through the various steps of the process and the extensive validation protocols followed to ensure that the new assay met a customer’s needs, generated data that was comparable to the previous tests, and performed as well in the customer’s lab as it did in the hands of MilliporeSigma scientists.

Development Team Leads to Success

Godeny also shared some interesting statistics from the custom assay development team, which has developed everything from single-plex to 48-plex xMAP assays. They’ve developed more than 50 new analytes and had a success rate better than 98 percent.

If you’re curious about custom assay services, we encourage you to take a few minutes to watch Godeny’s informative presentation.

Resources

METHODS Special Issue: Multiplexing Enables More Robust Surveillance for Avian Influenza

New assay developed for routine poultry monitoring

METHODS Special Issue: Multiplexing Enables More Robust Surveillance for Avian Influenza

In the journal Methods, scientists from Wageningen Bioveterinary Research report the development of a multiplex assay for avian influenza. The assay is built on Luminex’s Advances in Bead-Based Biomarker Detection [Methods xMAP Issue]

  • ® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP® Cookbook to Design Your Own Assays [Download]
  • METHODS Special Issue: Multiplexing Enables More Robust Surveillance for Avian Influenza

    • METHODS Special Issue: New Assays to Characterize Vaccination and Infection History

    Single-plex targets developed will become components for future multiplex assays

    Scientists in the Waterboer lab at the German Cancer Institute have developed the building blocks for a multiplex assay that will make it possible to quickly interrogate a person’s history of infection and vaccination. In a paper from Methods, the team reports developing individual assays for antibodies to several pathogens, noting that these assays could easily be combined into a multiplex assay in the future. These types of assays “are useful and versatile instruments” that can be utilized for “clinical diagnosis, personal risk evaluation, and seroepidemiological studies,” the scientists write.

    While the team makes it clear that the ultimate goal is to develop multiplex serology assays that cover antibodies for many pathogens to economize time, cost, and sample volume, they first had to expand the number of available single-pathogen targets using a bead-based system. To accomplish this, they used xMAP® Technology from Luminex.

    “We developed and validated bead-based pathogen-specific Monoplex Serology assays … to detect antibodies against Corynebacterium diphtheriae and Clostridium tetani toxins, rubella virus, and parvovirus B19,” they report, noting that each assay features a single antigen to improve the efficiency of future multiplexing. These assays “can be efficiently incorporated into larger multiplex serology panels,” the scientists add.

    Validation was performed using commercially available reference assays. Specificity, sensitivity, and other metrics were found to be acceptable for routine clinical use. This work provides a critical foundation for future multiplex development, and will streamline the characterization of vaccination and infection history for any individual.

    Resources

    METHODS Special Issue: New Assays to Characterize Vaccination and Infection History

    Webinar: Protein Conformational Analysis Goes High-Throughput with xMAP Technology

    Technique provides critical data for development of monoclonal antibodies and biosimilars

    At a 2018 xMAP® Connect user group meeting, Xing Wang, President of Array Bridge, spoke about how his team is using 30-minute presentation focuses on protein conformational analysis (PCA), a multiplex approach that replaces previous testing performed with ELISAs.

    The need for better tools to develop biologics — Wang’s talk centered on monoclonal antibodies — is clear from the rapidly growing market, he said, noting that by 2020 it is expected that more than 70 monoclonal antibody therapies will be on the market. Pharmaceutical and biotech companies are investing heavily in the development of these therapeutics, and existing products are eventually expected to be mirrored with new biosimilars.

    How PCA Helps

    A major challenge in that development, however, lies in understanding the three-dimensional conformation of a protein — including how that influences its biological function and its reaction to stressors. PCA offers a straightforward way to do this by using antibodies to recognize epitopes on the surface of the mAb, Wang said. It’s also highly sensitive and high-throughput, two major advantages over many other techniques.

    PCA has already been used in many biosimilar applications to the FDA and EU, including the first biosimilar mAb approved in the world with the highest standards of regulation. It has been used by the majority of biosimilar developers in the world with PCA data included in several successful approvals from US, EU, and other countries, and the technology has a number of potential applications, Wang said.

    Development of PCA

    The presentation also looked at how PCA was developed as Wang’s team shifted from sandwich-based ELISA tests to xMAP Technology for increased throughput. He highlighted results from real samples comparing both techniques to show that the data profiles are very similar, and also cited a publication from Bristol-Myers Squibb scientists using PCA for 48-fold higher throughput than ELISAs. The multiplex assay requires little sample and minimal hands-on time and is also very fast with high precision and broad dynamic range, Wang added.

    Finally, Wang reviewed data from analyses of immunogenicity and higher order structure correlation. These studies assess drug response and spot issues associated with immunogenicity, including human whole blood assay for cytokine release and complement activation, which explain why some people respond differently to the same treatment. The antibody arrays developed by Wang’s team generated a unique signature for each monoclonal antibody, producing sensitive results and detecting changes that may be missed by other bioassays and bioanalytical technologies.

    Related Content

    METHODS Special Issue: A Universal Protocol for Attaching Any Peptide to Dyed Magnetic Beads

    A new technique eliminates the use of monoclonal antibodies and is robust enough for field use

    [thumbnail]

    Monoclonal antibodies may soon become a thing of the past. While their high selectivity and affinity for their targets once made them the “gold standard” for detection applications, the time it takes to generate one and the variability in sensitivity has made the need for an alternative more prominent in recent years.

    This is the challenge scientists at the US Army Research Laboratory tackled and reported in a recent Methods publication.

    In the publication, authors Matthew Coppock and Dimitra Stratis-Cullum describe efforts to attach peptides to magnetic beads for potential use in the field. Their studies were performed using the Advances in Bead-Based Biomarker Detection [Methods xMAP Issue]

  • ® Cookbook to Design Your Own Assays” href=”http://info.luminexcorp.com/en-us/research/download-the-xmap-cookbook”>xMAP® Cookbook to Design Your Own Assays [Download]
  • METHODS Special Issue: A Universal Protocol for Attaching Any Peptide to Dyed Magnetic Beads