Generating Cytokines and Growth Factors for Functional Activity: Feasibility of Method Using MIF Protein.

IF 2.3 Q3 BIOCHEMICAL RESEARCH METHODS Methods and Protocols Pub Date : 2024-09-12 DOI:10.3390/mps7050072
Hiba Osmani, Ishrya Sharma, Shannon Moonah
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引用次数: 0

Abstract

Cytokines and growth factors are signaling molecules that regulate a variety of biological processes. Understanding their role is essential for basic research and clinical utilization. Thus, cytokines and growth factors are widely used throughout research labs in a significant number of applications. Additionally, genetic polymorphisms result in variant forms of cytokines and growth factors, which can alter their function. Becoming more common, researchers will need to generate these important proteins and their variants themselves in functional forms for activity studies. The expression systems used to generate these proteins can have a major impact on their function. In some instances, post-translational modifications are needed to produce a functionally active protein, which can only be conducted using eukaryotic expression systems. Ideally, for functional relevance, a human expression system should be used for human-related research and applications. Most human cell-based expression systems primarily use HEK (Human Embryonic Kidney) cells; however, relying on just one cell type can lead to several issues, considering the variety of proteins derived from various cell sources. Here, we provide a protocol to effectively and efficiently generate functional recombinant proteins, taking into consideration the diverse range of proteins from different cell types throughout the human body.

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产生细胞因子和生长因子的功能活性:使用 MIF 蛋白的方法的可行性。
细胞因子和生长因子是调节各种生物过程的信号分子。了解它们的作用对于基础研究和临床应用至关重要。因此,细胞因子和生长因子在研究实验室中被广泛应用。此外,基因多态性会导致细胞因子和生长因子的变异形式,从而改变它们的功能。研究人员需要以功能形式生成这些重要的蛋白质及其变体,以进行活性研究,这种情况正变得越来越普遍。用于生成这些蛋白质的表达系统会对其功能产生重大影响。在某些情况下,需要进行翻译后修饰才能生成具有功能活性的蛋白质,而这只能通过真核表达系统进行。理想情况下,与功能相关的人类相关研究和应用应使用人类表达系统。大多数基于人体细胞的表达系统主要使用 HEK(人胚胎肾)细胞;然而,考虑到从不同细胞来源获得的蛋白质种类繁多,仅仅依赖一种细胞类型可能会导致一些问题。在此,我们提供了一种有效、高效地生成功能性重组蛋白的方案,其中考虑到了来自人体不同细胞类型的各种蛋白质。
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
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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