Mini review for niche downstream processes

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-02-18 DOI:10.1016/j.pep.2025.106690

Tsutomu Arakawa , Daisuke Ejima , Yui Tomioka , Chiaki Sakuma , Teruo Akuta

引用次数: 0

Abstract

We review here several niche downstream purification processes that are not covered by other articles in this special issue. The first is the use of activated carbon to capture contaminants and clarify culture medium for purification of proteins, including antibodies. Flow-through operation of the activated carbon filter showed over 80 % recovery of antibodies with 10--fold reduction of host cell proteins and effective lipopolysaccharide and virus removal. The second is salt or arginine-tolerant column chromatography, in which the respective resins, such as hydroxyapatite and mixed-mode resins, can bind proteins in the presence of salt or arginine at high concentrations. Effective use of arginine was also suggested in size exclusion and affinity chromatography. The last is the isolation of proteins using gel electrophoresis and simple extraction procedure. These technologies offer simple and cost-effective methods for purifying proteins and protein complexes in the native state.

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我们在此回顾本特刊其他文章未涉及的几种利基下游纯化工艺。首先是使用活性炭捕获污染物并澄清培养基，以纯化包括抗体在内的蛋白质。活性炭过滤器的流过式操作表明，抗体的回收率超过 80%，宿主细胞蛋白减少了 10 倍，脂多糖和病毒也被有效去除。第二种是耐盐或耐精氨酸柱层析法，其中相应的树脂（如羟基磷灰石和混合模式树脂）可在高浓度盐或精氨酸存在的情况下结合蛋白质。在尺寸排阻和亲和层析中，精氨酸也得到了有效利用。最后一种是利用凝胶电泳和简单的提取程序分离蛋白质。这些技术为纯化原生状态的蛋白质和蛋白质复合物提供了简单而经济有效的方法。

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

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and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.