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

• Published in: Biochemical and biophysical research communications Vol. 512; no. 3; pp. 531 - 536
• Main Authors: Wu, Mengjuan, Tu, Zhui, Huang, Fengchun, He, Qinghua, Fu, Jinheng, Li, Yanping
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 07-05-2019
• Subjects: Cell Surface Display Techniques; Enzyme-Linked Immunosorbent Assay; Humans; Immunomagnetic beads; Immunomagnetic Separation; Lipopolysaccharide; Lipopolysaccharides - immunology; Nanobody; Phage display; Single-Domain Antibodies - immunology; Vibrio - immunology; Vibrio - isolation & purification; Vibrio fluvialis; Vibrio Infections - microbiology; Lipopolysaccharide; Vibrio fluvialis; Nanobody; Phage display; Immunomagnetic beads
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2019.03.104

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.
•Lipopolysaccharide was extracted as a target for panning nanobodies.•Anti-LPS nanobody for Vibrio fluvialis was obtained from naïve nanobody library.•Anti-LPS nanobody can be served as a capture target to enrich Vibriofluvialis.