摘要
制备抗幽门螺杆菌(Helicobacter pylori)尿素酶B(UreB)纳米抗体。用灭活的全幽门螺杆菌免疫羊驼,以重组蛋白UreB为靶分子,从噬菌体展示文库中筛选出能与重组蛋白UreB特异性结合的纳米抗体,构建原核表达载体,IPTG诱导表达,ELISA鉴定纳米抗体特异性。获得5×10^(7)cfu的噬菌体文库,将筛选的两个OD450值较高且序列不同的纳米抗体HU2-9和HU2-36分别进行表达SDS-PAGE鉴定均为可溶性且表达量分别为92和32 mg·L^(-1)。ELISA结果表明两个纳米抗体能特异性结合Hp菌体蛋白,并对尿素酶活性具有一定的抑制作用。成功淘选出6种独特序列的抗幽门螺杆菌UreB纳米抗体。
To prepare and characterize the nanobody against urease B(UreB)of Helicobacter pylori.The alpaca was immunized with the lysate of inactivated Hp.Using the recombinant protein UreB as target molecule,nanobodies that can specifically recognize recombinant protein UreB were screened from phage displayed immune libraries.Prokaryotic expression vector was constructed and induced by IPTG.Specificity of the nanobodies was confirmed by ELISA.A 5×10^(7) phage display library was obtained.The screened nanobodies HU2-9and HU2-36 with high OD450 in phage-ELISA and different sequences were recombinantly expressed.The SDS-PAGE analysis showed that the production was soluble and the production yield were 92 mg·L^(-1) and 32 mg·L^(-1),respectively.ELISA showed that production which was recombinantly expressed could bind to the lysate of Hp.The production could inhibit urease activity.nanobodies against urease B of Helicobacter pylori were successfully obtained.
作者
钟引凤
刘晓芳
刘贝
黄云祥
何庆华
李燕萍
涂追
刘琼
付金衡
ZHONG Yin feng;LIU Xiao fang;LIU Bei;HUANG Yun xiang;HUE Qing hua;LI Yan ping;TU Zhui;LIU Qiong;FUJin heng(School of Life Sciences,Nanchang University,Nanchang 330031,China;OAI Sino-German United Research Institute,Nanchang 330047,China;State Key Laboratory of Food Science and Technology,Nanchang 330047,China;School of Basic Medicine,Nanchang University,Nanchang 330031,China)
出处
《南昌大学学报（理科版）》
CAS
北大核心
2021年第1期41-47,共7页
Journal of Nanchang University(Natural Science)
基金
国家自然科学基金资助项目(31471648)。
