Department of Dermatology, University of California, Davis, Sacramento, CA, USA.
XLock Biosciences, LLC, West Allis, WI, USA; Boston Children's Hospital, Boston, MA, USA.
J Immunol Methods. 2023 Apr;515:113453. doi: 10.1016/j.jim.2023.113453. Epub 2023 Feb 28.
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.
一种新型工程化的 CCL20 二聚体（CCL20LD）与天然存在的趋化因子 CCL20 几乎相同，但可阻断 CCR6 介导的趋化作用，并为治疗银屑病和银屑病关节炎等疾病提供了一种新方法。需要定量检测 CCL20LD 血清水平的方法，以评估药代动力学参数，并评估药物输送、代谢和毒性。现有的 ELISA 试剂盒无法区分 CCL20LD 和天然趋化因子 CCL20WT（野生型单体）。在此，我们测试了几种现有的 CCL20 单克隆抗体，以能够鉴定出一种既能作为捕获抗体又能作为检测抗体（带有生物素标记）的克隆，从而能够特异性地高特异性检测 CCL20LD。使用重组蛋白进行验证后，使用 CCL20LD 选择性 ELISA 分析了接受 CCL20LD 治疗的小鼠的血液样本，证明了这种新型检测方法在用于银屑病疾病的生物制药先导化合物的临床前开发中的实用性。
