Expression and purification of Hepatitis B virus core antigen using Escherichia coli and its utilization for the diagnosis of Hepatitis B virus infections

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2023-11-16 DOI:10.1016/j.biologicals.2023.101726

Mohammed El-Mowafy , Mohamed El-Mesery , Mahmoud A.F. Khalil , Ahmed El-Mesery , Abdelaziz Elgaml

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引用次数: 0

Abstract

Hepatitis B virus (HBV) is responsible for most of the viral hepatitis worldwide. HBV is a partially double stranded DNA virus that is composed of four main open reading frames (ORFs) encoding its important antigens, namely hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg), HBV polymerase and hepatitis B X antigen (HBxAg). In this study, we report a successful method for the cloning and expression of HBcAg. The ORF of HBcAg was successfully amplified using polymerase chain reaction (PCR), cloned into the expression vector pRSET-B and transformed to Escherichia coli (E. coli) BL-21 (DE3) pLysS strain for protein expression. Successful expression of HBcAg was accomplished, in which an induced protein with a molecular weight of 24 kDa was obtained and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The produced HBcAg was successfully used for the diagnosis of HBV infected patient through detection of antibodies against HBcAg (anti-HBcAg) in the serum of the patient utilizing Western blotting. Overall, this study provides a simple, convenient and efficient protocol for the production of HBcAg that can be used as an important candidate to study the diagnosis and prognosis of HBV disease, as well as for understanding the epidemiological prevalence of HBV cases and production of anti-HBcAg.

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用大肠杆菌表达纯化乙型肝炎病毒核心抗原及其在乙型肝炎病毒感染诊断中的应用

乙型肝炎病毒(HBV)是世界上大多数病毒性肝炎的罪魁祸首。HBV是一种部分双链DNA病毒，由编码其重要抗原的四个主要开放阅读框(orf)组成，即乙型肝炎表面抗原(HBsAg)、乙型肝炎核心抗原(HBcAg)、HBV聚合酶和乙型肝炎X抗原(HBxAg)。在这项研究中，我们报道了一种成功的HBcAg的克隆和表达方法。利用聚合酶链反应(PCR)成功扩增HBcAg的ORF，克隆到表达载体pRSET-B中，转化到大肠杆菌(e.c oli) BL-21 (DE3) pLysS菌株中进行蛋白表达。成功表达了HBcAg，获得了分子量为24 kDa的诱导蛋白，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western blotting进行了证实。通过Western blotting检测患者血清中针对HBcAg的抗体(anti-HBcAg)，成功地将所制备的HBcAg用于HBV感染患者的诊断。总的来说，本研究为HBcAg的产生提供了一个简单、方便、高效的方案，可作为研究HBV疾病诊断和预后，以及了解HBV病例流行情况和抗HBcAg产生的重要候选方案。

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

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.