Circular dichroism spectroscopy in protein engineering and pharmaceutical development: Applications in structural characterization and quality assessment

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-10-08 DOI:10.1016/j.pep.2025.106826

Taiji Oyama, Satoko Suzuki, Ken-ichi Akao

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引用次数: 0

Abstract

Circular dichroism (CD) spectroscopy is a rapid and versatile method for assessing the higher-order structures of proteins and peptides. Far-UV CD enables quantitative evaluation of secondary structures, while near-UV CD provides sensitive fingerprints of tertiary organization. With advances in recombinant protein expression, CD spectroscopy has become widely used for the characterization of novel generated proteins. This review focuses on the applications of CD spectroscopy in protein engineering and pharmaceutical sciences. Examples include strategies for reliable measurements with limited sample quantities and quality assessments such as lot-to-lot comparisons and biosimilar evaluations. CD spectroscopy also serves as a valuable tool for detecting conformational changes associated with protein–protein and protein–drug interactions, as well as for evaluating proteins and peptides in membrane-mimetic environments. Obtaining reliable CD spectra requires careful selection of buffers, water-miscible solvents, and excipients. Here, we summarize their properties and propose practical criteria for their selection to ensure high-quality CD measurements.

查看原文本刊更多论文

圆二色光谱在蛋白质工程和药物开发中的应用：结构表征和质量评价。

圆二色光谱（CD）是一种快速和通用的方法，用于评估蛋白质和肽的高阶结构。远紫外CD可以定量评价二级结构，而近紫外CD提供三级组织的敏感指纹。随着重组蛋白表达技术的进步，CD光谱已被广泛用于鉴定新生成的蛋白质。本文综述了CD光谱在蛋白质工程和制药科学中的应用。例子包括有限样本量的可靠测量策略和质量评估，如批对批比较和生物类似药评估。CD光谱学也是检测与蛋白质-蛋白质和蛋白质-药物相互作用相关的构象变化以及评估膜模拟环境中的蛋白质和肽的有价值的工具。获得可靠的CD光谱需要仔细选择缓冲液、水混溶溶剂和赋形剂。在这里，我们总结了它们的特性，并提出了选择它们的实用标准，以确保高质量的CD测量。

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.