噬菌体展示系统的构建及其在疾病防控领域应用研究进展

doi:10.16431/j.cnki.1671-7236.2022.05.010

摘要/Abstract

摘要： 噬菌体展示技术作为目前应用广泛的展示抗体技术，逐渐成为生产基因工程抗体的重要工具。噬菌体展示技术是以噬菌体或噬菌粒为载体，通过将外源多肽基因整合到噬菌体基因中，以融合表达的形式将外源蛋白展示在噬菌体表面的分子生物学技术。近年来，噬菌体展示技术在抗体筛选领域的应用越来越广泛，与传统制备抗体的方法相比，噬菌体展示技术具有通量高、成本低、操作简单等特点，且通过该技术筛选得到的抗体不仅可在标签蛋白的辅助下进行选择和纯化，还可通过基因测序的方法得到单链抗体的完整基因序列。笔者首先对噬菌体抗体库进行分类，根据抗体的来源将噬菌体抗体库分为天然抗体库和免疫抗体库，通过免疫动物制备的抗体文库的特异性、抗体阳性率明显高于天然抗体文库;其次简述了噬菌体抗体库的构建流程，其中噬菌体表达载体的选择是展示技术的关键，整合了噬菌粒基因的辅助噬菌体侵染其特异性菌株，从而通过抗生素平板对该系统进行选择性筛选;最后讨论了噬菌体展示技术在疾病防控领域应用的研究进展，抗体的首次人源化使同源性抗体在临床上应用成为可能，后续用于预防和治疗病毒性疾病的动物同源性抗体的研发进一步证明了噬菌体展示技术用于生产诊断或潜在治疗试剂的能力。该综述主要聚焦于噬菌体展示系统的构建及其在疾病防控领域的应用，以期对后续通过噬菌体展示技术筛选单链抗体的应用提供指导。

关键词: 噬菌体展示技术; 融合表达; 单链抗体; 噬菌粒; 疾病防控

Abstract: As the most widely used antibody display technology,phage display technology has gradually become an important tool for the production of genetically engineered antibodies.Phage display technology is a molecular biology technology that uses phage or phagemid as a carrier to integrate exogenous polypeptide genes into phage genes and display the exogenous protein on the surface of the phage in the form of fusion expression.In recent years,phage display technology has been used more and more widely in the field of antibody screening.Compared with traditional methods for preparing antibodies,phage display technology has the characteristics of high throughput,low cost,and simple operation.And the antibodies screened by this technology can not only be selected and purified with the assistance of the tag protein,but also the complete gene sequence of the single-chain antibody can be obtained by the method of gene sequencing.This review first classified the phage antibody library.According to the source of the antibody,the phage antibody library could be divided into natural antibody library and immune antibody library.The specificity and positive rate of antibody libraries prepared from immunized animals were significantly higher than those of natural antibody libraries.Then the construction process of the phage antibody library was briefly described.The selection of the phage expression vector was the key to the display technology.The helper phage integrated with the phagemid gene infected its specific strains,so that the system could be selectively screened through antibiotic plates.Finally,the research progress of the application of phage display technology in the field of disease prevention and control was discussed.The first humanization of antibodies had made it possible for homologous antibodies to be used in clinical applications.Subsequent research and development of animal homologous antibodies for the prevention and treatment of viral diseases had proved the ability of phage display technology to produce diagnostic or potential therapeutic reagents.This review mainly focused on the construction of phage display system and its application in the field of disease prevention and control,in order to guide the subsequent screening of single-chain antibodies through this technology.

Key words: phage display technology; fusion expression; single chain antibody; phagemid; disease prevention and control

中图分类号:

侯伟. 噬菌体展示系统的构建及其在疾病防控领域应用研究进展[J]. 中国畜牧兽医, 2022, 49(5): 1688-1696.

HOU Wei. Research Progress on Construction of Phage Display System and Its Application in the Field of Disease Prevention and Control[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1688-1696.

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