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制备和构建嵌合型人源化广谱反应性抗体 10H10 针对蜱传脑炎病毒蛋白 E。

Preparation and Construction of Chimeric Humanized Broadly Reactive Antibody 10H10 to Protein E of Tick-Borne Encephalitis Virus.

机构信息

State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.

Federal Research Center for Fundamental and Translational Medicine, Novosibirsk, Russia.

出版信息

Bull Exp Biol Med. 2024 Oct;177(6):770-773. doi: 10.1007/s10517-024-06265-y. Epub 2024 Oct 23.

Abstract

A full-length humanized chimeric antibody 10H10ch that specifically interacts with the surface glycoprotein E of flaviviruses was obtained. To construct it, we used variable fragments of the heavy and light chains of the monoclonal antibody 10H10 that form the active center of the antibody and a fragment of the constant part of the heavy chain of the human IgG1 antibody. The resulting full-length chimeric humanized antibody 10H10ch specifically interacted with the E protein of flaviviruses pathogenic to humans, such as tick-borne encephalitis, Zika, West Nile, and dengue viruses. An immunochemical assessment of the interaction constants of the 10H10ch antibody with a panel of native and recombinant flavivirus antigens by ELISA and biolayer interferometry showed that the dissociation constant (Kd) of the chimeric antibody is in the nanomolar region and is comparable to that of the high-affinity mouse monoclonal antibody 10H10. The possibility of using the resulting chimeric humanized antibody 10H10ch for the diagnosis, prevention, and treatment of various flavivirus infections is discussed.

摘要

获得了一种与人源全长嵌合抗体 10H10ch 特异性相互作用的全长嵌合抗体,该抗体能特异性结合黄病毒表面糖蛋白 E。为了构建它,我们使用了形成抗体活性中心的单克隆抗体 10H10 的重链和轻链可变片段以及人 IgG1 抗体重链的恒定部分片段。所得全长嵌合人源化抗体 10H10ch 特异性地与人源致病性黄病毒,如蜱传脑炎、寨卡、西尼罗河和登革热病毒的 E 蛋白相互作用。通过 ELISA 和生物层干涉测量法对 10H10ch 抗体与一组天然和重组黄病毒抗原的相互作用常数进行免疫化学评估表明,嵌合抗体的解离常数(Kd)在纳摩尔范围内,与高亲和力的鼠源单克隆抗体 10H10 相当。讨论了用于诊断、预防和治疗各种黄病毒感染的所得嵌合人源化抗体 10H10ch 的可能性。

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