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

Autor: Wu X; Department of Dermatology, University of California, Davis, Sacramento, CA, USA., Clarke WR; XLock Biosciences, LLC, West Allis, WI, USA; Boston Children's Hospital, Boston, MA, USA., Koplinski CA; Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA; XLock Biosciences, LLC, West Allis, WI, USA., Peterson FC; Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA; XLock Biosciences, LLC, West Allis, WI, USA., Dwinell MB; Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI, USA., Wei G; USF Health Morsani College of Medicine, Tampa, FL, USA., Chao E; California Northstate University, College of Medicine, Elk Grove, CA, USA., Huynh M; Department of Dermatology, University of California, Davis, Sacramento, CA, USA., Yamada D; Department of Dermatology, University of California, Davis, Sacramento, CA, USA., Volkman BF; Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA; XLock Biosciences, LLC, West Allis, WI, USA., Hwang ST; Department of Dermatology, University of California, Davis, Sacramento, CA, USA. Electronic address: sthwang@ucdavis.edu.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2023 Apr; Vol. 515, pp. 113453. Date of Electronic Publication: 2023 Feb 28.
DOI: 10.1016/j.jim.2023.113453
Abstrakt: 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.
Competing Interests: Declaration of Competing Interest STH, BFV, MBD, WFC, FCP, and CAK have financial interest in and are officers of XLock Biosciences, which produces the CCL20LD.
(Copyright © 2023. Published by Elsevier B.V.)
Databáze: MEDLINE
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