两步纯化HBV中间蛋白，应用离子交换色谱法提高蛋白均匀性。

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-06-28 DOI:10.1016/j.pep.2025.106765

Nadia Hadiji-Abbes, Nihel Ammous-Boukhris, Istabrak Borchani-Chabchoub, Ali Gargouri, Raja Mokdad-Gargouri

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引用次数: 0

摘要

重组乙型肝炎表面抗原（PreS2-S）在酿酒酵母中产生，并被评价为乙型肝炎病毒（HBV）感染的候选疫苗。重组病毒样颗粒（vlp）是首个获批的基于vlp的疫苗。我们之前的研究表明，利用毕赤酵母和酿酒酵母，以及优化的VLP提取方法，可以在细胞内高产出重组HBsAg。在本研究中，我们旨在简化重组M蛋白（PreS2 + S）的纯化和表征。为了实现这一目标，我们采用了两步纯化过程，从尺寸排除色谱开始，然后是离子交换色谱，它具有有效分离中颗粒和大颗粒的优势。纯化的重组M蛋白成功地揭示了VLPs的存在，这是抗原性所必需的。

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Purification of the HBV middle protein by two simple steps, application of ion exchange chromatography to improve protein homogeneity

Recombinant hepatitis B surface antigen (PreS2-S) was produced in the yeast Saccharomyces cerevisiae and evaluated as a vaccine candidate against Hepatitis B Virus (HBV) infection. Recombinant virus-like particles (VLPs) were the first VLP-based vaccines to be approved. Our previous studies demonstrated high intracellular production of recombinant HBsAg using Pichia pastoris and Saccharomyces cerevisiae, along with optimized VLP extraction methods. In the present study, we aim to simplify the purification and characterization of recombinant M protein (PreS2 + S). To achieve this, we employed a two-step purification process, beginning with size exclusion chromatography followed by ion exchange chromatography that has the advantage of effectively separate the medium and large particles. The purified recombinant M protein successfully revealed the presence of VLPs, which are essential for antigenicity.

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.