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

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 Luminex® 200™ System since “Luminex instruments are of high interest because they can be found in some mobile lab settings and are more sensitive compared to other similar devices.”

As the scientists note, peptide-based reagents have become an appealing alternative to monoclonal antibodies for biological detection. “Peptides are advantageous due to their high thermal stabilities, which could help eliminate cold chain transport, improve long-term storage, and allow at-point detection in austere environments. Peptides also allow on-demand scalability,” they write. Using peptides in a multiplex system could permit high-throughput interrogation of samples, even in remote locations or under suboptimal lab conditions.

In the publication, Coppock and Stratis-Cullum describe a method for “the attachment of any peptide to dyed magnetic microspheres, regardless of peptide length, size, or sequence.” The technique makes use of single-arm or multi-arm PEG linkers. The authors add that “the method exploits ‘click’ chemistry with short reaction times in a mixed organic/water system for simultaneous selective surface functionalization and reduction of microsphere dye leaching.” They demonstrated the method using a Streptavidin binding peptide and biotin, among other examples.

Further development of this method could expand its use to other types of reagents and microspheres, the authors note.

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

Now Available: Custom Assay Services from Luminex

From full development to providing guidance, we can help you get to a functional assay—faster.

Now Available: Custom Assay Services from Luminex

The Luminex team is pleased to announce the launch of our custom development service, LuminexPLORE Lab, for assays based on xMAP® Technology. For more than 20 years, we have collaborated with customers and partners around the world in designing and developing high-quality, high-performing assays. Now, we are able to offer even more support and guidance for your research needs. This service will be run by Jackie Surls, our Development and Applications Scientist.

Many xMAP Technology users rely on published literature to design and develop new assays, and with good reason—there are vast resources available from peer-reviewed journals demonstrating great advances with xMAP-based assays. But not everyone has the time or bandwidth to wade through dozens of publications to find just the right method for their specific project.

We know that our customers are more than qualified to build their own assays. We have the most capable user base out there! But for those who simply don’t have time to design a new assay, or to fine-tune one that is already in development, our custom service group has you covered.

Our team has more than two decades of experience designing and developing xMAP-based assays, as well as advanced immunoassays and genomic assays. We have expertise in infectious disease, immunology, cell signaling, toxicity, allergy testing, metabolism, serology, and much more. Already, we’ve worked on several service projects involving full assay development, as well as custom coupling. We have expertise in a broad range of services:

  • Assay development
  • Assay optimization of existing homebrew xMAP assay
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  • Assay conversion
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Whether you are a new assay developer or trying to accelerate your project, our custom services program can help your team reach its goals—faster.

Please note: This service is for Research Use Only.

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Now Available: Custom Assay Services from Luminex

METHODS Special Issue: Modifying the Luminex Workflow for Hazardous Pathogen Research

New protocol allows scientists to create assays in containment labs and perform analysis elsewhere

Scientists in the UK have developed a new protocol that utilizes Luminex multiplex assays even in specialized laboratories where space for instruments is limited. A recent hazardous pathogen study has broad implications for any research team, especially those facing space limitations.

In a paper recently published in METHODS, Stuart Dowall, Victoria Graham, and Roger Hewson from Public Health England (PHE), along with Tom Fletcher from the Liverpool School of Tropical Medicine, describe the protocol they established to perform multiplex assays in specialized Containment Level 4 (CL4) facilities, which are designed for the most dangerous pathogens.

These laboratories differ from Biosafety Level 4 (BSL-4) facilities because “work is conducted in class III microbiological safety cabinets for primary containment instead of using positive pressure suits,” which rely on HEPA-filtered air in a positively pressurized microenvironment to protect researchers from the pathogens in their laboratory space, the authors report. “This presents unique challenges associated with the physical restrictions of working in a limited space, and prohibits the use of many techniques and specialized equipment.” As a result, research labs require innovative solutions to perform multiplex assays.

Multiplex assays are paving the way for improved pathogen research

Because multiplex immunoassays make a real difference in scientists’ ability to investigate the mechanisms of disease induction, the UK team came up with a different approach to perform these critical assays. Their idea was simple: what if they could prepare the Luminex assay within the containment lab’s safety cabinets, but then actually run it somewhere else?

The paper reports the validation of this method, which succeeded thanks to a fixation step. “We have developed an approach in which the Luminex assay is conducted within the CL4 laboratory, and a formalin-fixation stage is introduced to allow for analysis to be undertaken outside of containment,” the scientists write. After fixation, assay plates are loaded into a bag, sealed, and transported to another laboratory for analysis. In this study, an analysis was performed within 24 hours of fixation on a Luminex MAGPIX® System.

“Our data demonstrate that Luminex is an applicable tool for use at CL4 and that assays can be run reliably to generate reproducible, standardized data across different plates and individual experiments,” the scientists conclude. “This approach will be utilized at PHE … to determine the contribution of immunological markers in disease pathogenicity of viral hemorrhagic fever.” The research team highlighted that following this protocol, Luminex multiplex assays can be performed safely and effectively on CL4 pathogens.

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METHODS Special Issue: Modifying the Luminex Workflow for Hazardous Pathogen Research