Screening and initial identification of human anti-vibrio vulnificus haemolysin ScFv

Abstract

Objective To screen specific human antibodies against Vibrio vulnificus hemolysin ( VVH ) from natural single stranded large capacity phage antibody library and to perform initial identification of the antibodies, for the treatment of vibrio vulnificus infection. Methods The specific antibody against purified VVH was screened from large natural phage antibody library by 4 rounds ofadsorption-elution-amplificationscreening process. Soluble ScFvs were prepared through infecting E coli. HB2151 and inducing expression with IPTG. The antigen binding activity and DNA sequence of the antibodies against VVH were identified by ELISA and DNA fingerprint analysis. Results After 4 rounds of panning, 22 positive clones with specific binding ability to VVH were obtained, of which 8 obtained soluble expressions. DNA sequence analyses showed that the variable region genes of the clones belonged to different subgroups of immunoglobulin gene family. Conclusions The acquirement of human anti-VVH ScFv from large phage antibody library has laid a good foundation for the treatment of vibrio vulnificus infection. Key words: Large phage library; Vibrio vulnificus; Haemolysin protein; Soluble antibody