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制备及单克隆抗体针对乙型肝炎病毒聚合酶的功能评价。

Preparation and functional evaluation of monoclonal antibodies targeting Hepatitis B Virus Polymerase.

机构信息

Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology , Wuhan, Hubei Province, China.

School of Life Science & School of Public Health; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics; National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University , Xiamen, China.

出版信息

Virulence. 2021 Dec;12(1):188-194. doi: 10.1080/21505594.2020.1869391.

Abstract

HBV pol plays a critical role in the replication of hepatitis B virus (HBV). Previous studies conducted on HBV pol have produced limited evidence on HBV pol expression due to the lack of effective detection methods. The present study used the HBV pol (159-406 aa) protein as a target to screen for specific monoclonal antibodies that recognize HBV pol and subsequently evaluate their diagnostic and therapeutic value. Four antibodies (P3, P5, P12, P20) against HBV pol were obtained. Among them, the P20 antibody indicated optimal binding with HBV pol as demonstrated by Western blotting (WB) in a cell model transfected with the HBV genome. We also expressed P5 and P12 antibodies in mouse liver cells by transfection and the results indicated significant antiviral effects caused by these two antibodies especially P12. In summary, the present study established an antibody which was denoted P20. This antibody can be used to detect HBV pol expression by four HBV genomes via WB analysis. In addition, the antibody denoted P12 could exert antiviral effects via intracellular expression, which may provide a promising approach for the treatment of chronic hepatitis B.

摘要

HBV pol 在乙型肝炎病毒 (HBV) 的复制中起着关键作用。由于缺乏有效的检测方法,之前对 HBV pol 的研究仅提供了有限的 HBV pol 表达证据。本研究使用 HBV pol(159-406 aa)蛋白作为靶标,筛选出特异性识别 HBV pol 的单克隆抗体,并评估其诊断和治疗价值。获得了针对 HBV pol 的四种抗体(P3、P5、P12、P20)。其中,P20 抗体在转染 HBV 基因组的细胞模型中通过 Western blot(WB)显示出与 HBV pol 的最佳结合。我们还通过转染在小鼠肝细胞中表达了 P5 和 P12 抗体,结果表明这两种抗体特别是 P12 具有显著的抗病毒作用。总之,本研究建立了一种抗体,称为 P20。该抗体可通过 WB 分析检测四种 HBV 基因组中 HBV pol 的表达。此外,通过细胞内表达的 P12 抗体可以发挥抗病毒作用,这可能为慢性乙型肝炎的治疗提供一种有前途的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1299/7834045/6e1f584dc7fb/KVIR_A_1869391_F0001_OC.jpg

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