Faculty Collaboration Database

A modified ELISA assay differentiates CCL20 locked dimers from wild-type monomers. J Immunol Methods 2023 Apr;515:113453

Date

03/03/2023

Pubmed ID

36863695

Pubmed Central ID

PMC10715733

DOI

10.1016/j.jim.2023.113453

Scopus ID

2-s2.0-85150034077 (requires institutional sign-in at Scopus site)

Abstract

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.

Author List

Wu X, Clarke WR, Koplinski CA, Peterson FC, Dwinell MB, Wei G, Chao E, Huynh M, Yamada D, Volkman BF, Hwang ST

Authors

Michael B. Dwinell PhD Director, Professor in the Microbiology and Immunology department at Medical College of Wisconsin
Francis C. Peterson PhD Professor in the Biochemistry department at Medical College of Wisconsin
Brian F. Volkman PhD Professor in the Biochemistry department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Antibodies, Monoclonal
Chemokine CCL20
Chemotaxis
Enzyme-Linked Immunosorbent Assay
Mice
Psoriasis