A modified ELISA assay differentiates CCL20 locked dimers from wild-type monomers

• Published in: Journal of immunological methods Vol. 515; p. 113453
• Main Authors: Wu, Xuesong, Clarke, William R., Koplinski, Chad A., Peterson, Francis C., Dwinell, Michael B., Wei, Grace, Chao, Ellen, Huynh, Mindy, Yamada, Daisuke, Volkman, Brian F., Hwang, Samuel T.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2023
• Subjects: Animals; Antibodies, Monoclonal - therapeutic use; Chemokine CCL20 - genetics; Chemotaxis; Cytokine; ELISA; Enzyme-Linked Immunosorbent Assay; Locked dimer; Mice; Psoriasis; Psoriasis - drug therapy; Psoriasis - metabolism; IL23MC; Psoriasis; Locked dimer; CCL20; Cytokine; CCL20LD; ELISA
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2023.113453

A novel engineered CCL20 locked dimer (CCL20LD) is nearly identical to the naturally occurring chemokine CCL20 but blocks CCR6-mediated chemotaxis and offers a new approach to treat the diseases of psoriasis and psoriatic arthritis. Methods for quantifying CCL20LD serum levels are needed to assess pharmacokinetics parameters and evaluate drug delivery, metabolism, and toxicity. Existing ELISA kits fail to discriminate between CCL20LD and the natural chemokine, CCL20WT (the wild type monomer). Herein, we tested several available CCL20 monoclonal antibodies to be able to identify one clone that can be used both as a capture and a detection antibody (with biotin-labeling) to specifically detect CCL20LD with high specificity. After validation using recombinant proteins, the CCL20LD-selective ELISA was used to analyze blood samples from CCL20LD treated mice, demonstrating the utility of this novel assay for preclinical development of a biopharmaceutical lead compound for psoriatic disease.