Panning anti-LPS nanobody as a capture target to enrich Vibrio fluvialis

Mengjuan Wu; Zhui Tu; Fengchun Huang; Qinghua He; Jinheng Fu; Yanping Li

doi:10.1016/j.bbrc.2019.03.104

Panning anti-LPS nanobody as a capture target to enrich Vibrio fluvialis

Biochem Biophys Res Commun. 2019 May 7;512(3):531-536. doi: 10.1016/j.bbrc.2019.03.104. Epub 2019 Mar 22.

Authors

Mengjuan Wu¹, Zhui Tu¹, Fengchun Huang², Qinghua He¹, Jinheng Fu¹, Yanping Li³

Affiliations

¹ 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.
² State Key Laboratory of Food Science and Technology, Nanchang University, No.235 Nanjing East Road, Nanchang, 330029, China.
³ 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. Electronic address: liyanping@ncu.edu.cn.

PMID: 30905409
DOI: 10.1016/j.bbrc.2019.03.104

Abstract

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.

Keywords: Immunomagnetic beads; Lipopolysaccharide; Nanobody; Phage display; Vibrio fluvialis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Surface Display Techniques
Enzyme-Linked Immunosorbent Assay
Humans
Immunomagnetic Separation
Lipopolysaccharides / immunology*
Single-Domain Antibodies / immunology*
Vibrio / immunology*
Vibrio / isolation & purification*
Vibrio Infections / microbiology

Substances

Lipopolysaccharides
Single-Domain Antibodies

Supplementary concepts

Vibrio fluvialis