Multiplexing offers cost and time savings for generating critical kinetic data
Across the drug discovery and development process, understanding the kinetics of how antibodies bind is essential. This information is important for antibody selection, quality control processes, assay development for analyte detection, and much more.
Conventional techniques for generating kinetic data on antibodies tend to be tedious and time-consuming. Each antibody must be tested individually. For such important data, having a higher-throughput alternative would be ideal.
A new way to generate kinetic data on antibodies
Recently, scientists at the Fraunhofer Institute who study bioanalytics and bioprocessing (in the cell therapy and immunology division) evaluated a multiplex method for antibody kinetics. The Germany-based team is responsible for analyte detection in human health applications ranging from food contaminants and disease markers to drug abuse indicators and more. Led by Harald Seitz, head of the biomarker validation and assay development department, the team also included post-doc Sandra Muekusch and PhD student Timo Ramm. “The main aim of my work is trying to analyze all kinds of different samples,” says Seitz. “That involves a lot of quality control and other elements that are essential for assay development.”
For this project, they used the Luminex FLEXMAP 3D® System with bead-based xMAP® Technology capable of detecting up to 500 analytes in a single sample. Their study offers the first demonstration of how xMAP multiplexing can be used to generate kinetic data on antibody binding.
xMAP Technology passes the test
The scientists evaluated the xMAP-based approach by incubating different concentrations of a primary antibody with a fixed concentration of a secondary antibody, as well as direct labeling of the primary antibody. They calculated EC50 (effective dose, when 50 % of binding occurred) as the KD (binding affinity) value and compared the outcome to values obtained through conventional methods for measuring antibody binding kinetics. The work was based on peptides developed by Muekusch for phosphorylation studies.
“We compared Luminex to the gold standard of SPR, and the data looked good.”
“I had worked with Luminex before, and it was quite easy.” The team notes that Luminex results would be particularly useful for ranking antibody binding kinetics, allowing researchers to quickly identify the best ones and move ahead to assay development.
The comparable results indicate that xMAP multiplexing technology is an efficient, high-throughput alternative for generating essential data about antibody binding patterns. xMAP Technology works with extremely small sample volumes, making it simpler and more cost-effective to test antibody binding for drug discovery and other applications.
Learn more about this project in our new tech note
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