白鲢肌原纤维结合型丝氨酸蛋白酶单域抗体文本库构建及淘选

Construction and Panning of Single Domain Antibody Library against Silver Carp Myofibrill-Bound Serine Protease

DOI：10.3969/j.issn.1673-1689.2020.03.012

中文关键词: 肌原纤维结合型丝氨酸蛋白酶 铰口鲨 单域抗体 噬菌体展示技术

英文关键词: myofibril-bound serine proteinase,nurse shark,single-domain antibody（sdAbs）,phage-display technology

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中文摘要:

鱼肉中的肌原纤维结合型丝氨酸蛋白酶（myofibril-bound serine proteinase，MBSP）在加工过程中可降解肌原纤维，引起鱼糜制品的凝胶劣化。一种新型特异性高的鲨源抗体——单域抗体（single-domain antib odies,sdAbs），能直接与抗原结合并抑制酶的活性。研究主要完成了针对 MBSP 单域抗体制备的前期工作：文本库建立及淘选过程，以克隆表达的 MBSP 为抗原免疫铰口鲨鱼（nurse shark，Ginglymostoma cirratum），构建鲨鱼单域抗体文本库，并从中淘选针对 MBSP 的特异性单域抗体噬菌粒，最终得到 31 个能与 MBSP 特异性结合的噬菌粒，为获得抗 MBSP 单域抗体鉴定了基础，以期抑制或消除 MBSP 的活性，解决现有鱼糜制品凝胶劣化的问题。

英文摘要:

Myofibril-bound serine proteinase（MBSP）in fish can degrade myofibrils during processing，resulting in gel deterioration of surimi products. A new highly specific shark-derived antibody，single-domain antibodies（sdAbs)，binds directly to antigens and inhibits enzyme activity. In this paper, nurse shark (Ginglymostoma cirratum) was immunized with expressed MBSP to construct the library of single domain antibody, and 31 phagemids specific to MBSP were panned from the library. The results could provide the basis for the expression and identification of specific single domain antibody which was expected to overcome the gel deterioration of the surimi products by inhibiting or eliminating the activity of MBSP.

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