State Key Laboratory of Food Science and Technology, Nanchang University, No.235 Nanjing East Road, Nanchang, 330029, China; Sino-German Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang, 330029, China; School of Food Science and Technology, Nanchang University, No.999Xuefu Avenue, Nanchang, 330031, China.
State Key Laboratory of Food Science and Technology, Nanchang University, No.235 Nanjing East Road, Nanchang, 330029, China.
Biochem Biophys Res Commun. 2019 May 7;512(3):531-536. doi: 10.1016/j.bbrc.2019.03.104. Epub 2019 Mar 22.
Vibrio fluvialis is considered as a human pathogen in developing countries. This bacterium is widely distributed in seawater and harbors that contains traces of salt. V. fluvialis can cause human enteritis and diarrhea, which has broken out at a global scale. Lipopolysaccharide (LPS) is a key bacterial antigen used to classify V. fluvialis serogroups. In this research, phage display technology was adopted to isolate nanobodies from a naïve phage library by using LPS as the target antigen. The isolated nanobody was tested in LPS ELISA and bacterial enzyme-linked immunosorbent assay Nanobody V23 had a high affinity toward the pathogen and was utilized to synthesize immunomagnetic beads for the enrichment of V. fluvialis. The capture efficiency of the immunomagnetic beads against V. fluvialis was 90.7 ± 3.2% (N = 3) through the plate-counting method. We generated a high-affinity nanobody against LPS from V. fluvialis and developed a rapid method of enriching V. fluvialis by using immunomagnetic beads.
弗氏弧菌被认为是发展中国家的人类病原体。这种细菌广泛分布在海水中和含有盐分的港湾中。弗氏弧菌可引起人类肠炎和腹泻，在全球范围内爆发过。脂多糖 (LPS) 是一种用于对弗氏弧菌血清群进行分类的关键细菌抗原。在这项研究中，采用噬菌体展示技术，以 LPS 作为靶抗原，从天然噬菌体文库中分离纳米抗体。分离出的纳米抗体在 LPS ELISA 和细菌酶联免疫吸附试验中进行了测试。纳米抗体 V23 对病原体具有高亲和力，并用于合成免疫磁珠，以富集弗氏弧菌。通过平板计数法，免疫磁珠对弗氏弧菌的捕获效率为 90.7±3.2%（N=3）。我们从弗氏弧菌中产生了一种针对 LPS 的高亲和力纳米抗体，并开发了一种使用免疫磁珠快速富集弗氏弧菌的方法。
