Development of a bridging ELISA for detection of antibodies against ZV0203 in cynomolgus monkey serum

IF 1.3 4区医学 Q4 PHARMACOLOGY & PHARMACY

Journal of pharmacological and toxicological methods Pub Date : 2023-01-01 DOI:10.1016/j.vascn.2022.107210

Tingting Liu , Yongliang Tong , Jinli Gao , Wenjing Fang , Junliang Wu , Xionghua Peng , Xiaotian Fan , Xiaoqiang Chen , Jianhua Sun , Sheldon Cao , Zhaohui Li , Likun Gong , Qiuping Qin , Dianwen Ju

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引用次数: 2

Abstract

Immunogenicity has been a major concern in the safety evaluation of therapeutic proteins. The assessment of the unwanted immunogenicity of the therapeutic proteins performed in animals prior to clinical trials has been a regulatory requirement. In preclinical studies of therapeutic proteins, cynomolgus monkeys are usually the most relevant animal species. ZV0203, a recombinant humanized anti-human epidermal growth factor receptor 2 monoclonal antibody covalently bound to a cytotoxic drug (Duo-5), possesses a novel format of antibody drug conjugates. In this study, we reported the development, validation, and application of a bridging enzyme-linked immunosorbent assay for the detection of antibodies against ZV0203 in cynomolgus monkey serum. Drug interference at low positive control (18.0 ng/mL) and high positive control (130 ng/mL) of anti-ZV0203 antibodies was not observed when ZV0203 concentration is below 1.74 μg/mL and 1.49 μg/mL, respectively. In addition, no interference was found from mouse IgG1, but interference was observed with human IgG1. No effect of hemolysis was found on the analysis results of the testing samples present in 100% pooled rabbit serum containing 2% (V/V) erythrocyte hemolysates. Besides, spiked anti-ZV0203 antibody in rabbit serum was stable after 5 freeze/thaw cycles. The results showed that the method is suitable for the detection of anti-ZV0203 antibodies in cynomolgus monkey serum. The assay was also successfully applied in the repeated dose study of ZV0203.

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建立一种桥接ELISA检测食蟹猴血清ZV0203抗体的方法

免疫原性一直是治疗性蛋白安全性评价的一个主要问题。在临床试验之前，对治疗性蛋白在动物身上进行的不必要的免疫原性评估是一项监管要求。在治疗性蛋白的临床前研究中，食蟹猴通常是最相关的动物物种。z0203是一种与细胞毒性药物(Duo-5)共价结合的重组人源化抗人表皮生长因子受体2单克隆抗体，具有一种新型的抗体药物偶联物。在这项研究中，我们报道了桥接酶联免疫吸附法的开发、验证和应用，用于检测食蟹猴血清中ZV0203抗体。当ZV0203浓度分别低于1.74 μg/mL和1.49 μg/mL时，抗ZV0203抗体低阳性对照(18.0 ng/mL)和高阳性对照(130 ng/mL)无药物干扰。此外，小鼠IgG1未发现干扰，但对人IgG1有干扰。在含有2% (V/V)红细胞溶血物的100%混合兔血清中，未发现溶血对检测样品的分析结果有影响。经5次冻融循环后，兔血清中抗zv0203抗体稳定存在。结果表明，该方法适用于食蟹猴血清中抗zv0203抗体的检测。该方法也成功地应用于ZV0203的重复给药研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of pharmacological and toxicological methods PHARMACOLOGY & PHARMACY-TOXICOLOGY

CiteScore

3.60

自引率

10.50%

发文量

审稿时长

26 days

期刊介绍： Journal of Pharmacological and Toxicological Methods publishes original articles on current methods of investigation used in pharmacology and toxicology. Pharmacology and toxicology are defined in the broadest sense, referring to actions of drugs and chemicals on all living systems. With its international editorial board and noted contributors, Journal of Pharmacological and Toxicological Methods is the leading journal devoted exclusively to experimental procedures used by pharmacologists and toxicologists.