Development of immunological reagents for the identification of biomarkers of inflammation in the common marmoset

Marmosets (MAR, Callithrix jacchus) are widely studied nonhuman primates in a variety of preclinical biomedical research fields. However, the scarcity of available immunological reagents poses a significant limitation for these models. Researchers have now successfully developed a pair of monoclonal antibodies (mAbs) that can be utilized in immunoassays to identify and quantify biomarkers of inflammation, including C-reactive protein (CRP), CXCL-10 (IP-10), interleukin (IL)-4, IL-6, and IL-10.

To create these mAbs, recombinant MAR protein variants were produced in mammalian cells and purified, subsequently used to immunize mice. Hybridoma clones were selected based on their ability to bind these recombinant proteins. The researchers then tested all possible combinations of these mAbs to determine their effectiveness in recognizing their targets within MAR-derived samples.

These MAR samples comprised the supernatant of peripheral blood mononuclear cells (PBMCs) exposed ex vivo to various stimuli, as well as plasma samples from animals stimulated in vivo with a low-dose intravenous LPS inoculation.

While many of the mAbs that recognized the recombinant proteins failed to bind to the MAR-derived homologs, the researchers identified MAR-derived reactive mAbs with optimal sensitivity and signal-to-noise ratios for immunoassays. These assays encompassed Luminex multiplexing assays, Enzyme-Linked Immunosorbent Assay (ELISA), and Enzyme-Linked Immunosorbent Spot (ELISpot) assays. By providing these immunoreagents and assays, the researchers aim to enhance the translational value of various MAR biomedical models.