食品科学2017，Vol.38Issue(6)：111-116,6.DOI:10.7506/spkx1002-6630-201706017
弧菌外膜蛋白OmpK单克隆抗体的制备及其特性
Preparation and Characterization of Monoclonal Antibodies against Vibrio Outer Membrane Protein OmpK
摘要
Abstract
This study aimed to prepare and characterize monoclonal antibodies (Mab) against outer membrane protein K (OmpK) of Vibrio.We injected Balb/c mice with OmpK of the Vibrio parahaemolyticus wild-type strain B expressed by a prokaryotic expression system.Spleen cells from the immunized mice were fused with SP2/0 tumor cells.Then the hybrid cell lines which can stably secrete monoclonal antibody were screened out by using the limited dilution method and indirect enzyme linked immunosorbent assay (ELISA) method.Ascites was produced in the mice and then purified by saturated ammonium sulfate precipitation and Protein G affinity chromatography.Finally,we obtained two hybridoma cell lines namely OmpK-Mab-4B7 and OmpK-Mab-3C5,which can stably secrete anti OmpK monoclonal antibody.The Mab subtypes secreted by both hybrid tumor cell lines were IgG1 and their titers reached 1:128 000.The sensitivity IC50 of 3C5 and 4B7 antibodies were 2.5 and 5.0 μg/mL,respectively.The results of Western blotting showed that the Mab could combine with the outer membrane proteins of 12 strains Vibrioparahaemolyticus (A,B,C,D,E,F,G,H,I,J,ATCC17802,and ATCC33847),3 (A,B,and C) of 4 Hbrio alginolyticus strains and 1 Vibrio anguillarum strain (MVM).The monoclonal antibodies can be used for basic research and rapid detection of OmpK.关键词
弧菌/弧菌外膜蛋白K/单克隆抗体Key words
Vibrio/outer membrane protein K/monoclonal antibody分类
轻工纺织引用本文复制引用
李杰,董庆利,刘箐,丁承超,翟续昭,王广彬,刘武康,谢曼曼,曾海娟,王淑娟,孙静娟..弧菌外膜蛋白OmpK单克隆抗体的制备及其特性[J].食品科学,2017,38(6):111-116,6.基金项目
上海市科技创新行动计划项目(15395810900) （15395810900）
上海理工大学研究生创新基金资助项目 （）
上海理工大学微创励志创新基金项目 （）
