Screening and initial identification of human anti-vibrio vulnificus haemolysin ScFv
- Author:
- No author available
- Journal Title:
- Chinese Journal of Nautical Medicine and Hyperbaric Medicine
- Issue:
- 2
- DOI:
- 10.3760/cma.j.issn.1009-6906.2016.02.014
- Key Word:
- 大容量噬菌体抗体库;创伤弧菌;溶血素蛋白;可溶性抗体;Large phage library;Vibrio vulnificus;Haemolysin protein;Soluble antibody
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 of " adsorption-elution-amplification" screening 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.
- [1]乔媛媛,王琰,陈晓穗,等.大容量噬菌体抗体库的构建及鉴定[J].2004,(3).
- [2]王欲晓,乔媛媛,周丽君,等.从大容量抗体库中筛选抗TNF-α人单链抗体[J].2008,(9).
- [3]付凯飞,马骢,郭建巍,等.抗创伤弧菌单克隆抗体杂交瘤细胞系的建立及特性鉴定[J].2012,(3).
- [4]Cho-Rok Jung,Min-Jung Park,Moon-Soo Heo.Immunization with Major Outer Membrane Protein of Vibrio vulnificus Elicits Protective Antibodies in a Murine Model[J].2005,43(5).
- [5]Li ZJ,Lei LC,Han WY,etc.Klebsiella pneumoniae MrkD adhesin-mediated immunity to respiratory infection and mapping the antigenic epitope by phage display library[J].2009,46(3).
- [6]R W, Parker,D H, Lewis.Sandwich enzyme-linked immunosorbent assay for Vibrio vulnificus hemolysin to detect V. vulnificus in environmental specimens.[J].1995,61(2).
- [7]Tae Hee Lee,Mi Hyun Kim,Chang-Seop Lee,etc.Protection against Vibrio vulnificus infection by active and passive immunization with the C-terminal region of the RtxA1/MARTXVv protein[J].2014,32(2).
- [8]Barrera R,Larsen JL,Amaro C,etc.Field testing of a vaccine against eel diseases caused by Vibriovulnificus[J].2001,45(3).
- [9]Lee,T.H.,Cha,S.-S.,Lee,C.-S.,etc.Monoclonal antibodies against Vibrio vulnificus RtxA1 elicit protective immunity through distinct mechanisms[J].2014,82(11).
- [10]van Egmond M,Lewis MJ,Pleass RJ,etc.A novel human IgA monoclonal antibody protects against tuberculosis.[J].2011,186(5).
- [11]Xian-Chun Tang,Agnihothram, Sudhakar S.,Yongjun Jiao.Identification of human neutralizing antibodies against MERS-CoV and their role in virus adaptive evolution.[J].2014,111(19).
- [12]Jones, Benjamin,Burkhart, Joshua,Sonpavde, Guru.Danger on a Half Shell: Vibrio vulnificus Septicemia[J].2015,128(5).
- [13]Hong, Zhao,Lichen, Xu,Huihui, Dong,etc.Correlations between Clinical Features and Mortality in Patients with Vibrio vulnificus Infection.[J].2015,10(8).
- [14]Ornnuthchar Poungpair,Kunan Bangphoomi,Prapaipit Chaowalit,etc.Generation of human single-chain variable fragment antibodies specific to dengue virus non-structural protein 1 that interfere with the virus infectious cycle[J].2014,6(2).
- [15]Kaus,K.,Lary,J.W.,Cole,J.L.,etc.Glycan specificity of the vibrio vulnificus hemolysin lectin outlines evolutionary history of membrane targeting by a toxin family[J].2014,426(15).
- [16]Angela, Cardaci,Salvatore, Papasergi,Angelina, Midiri,etc.Protective activity of Streptococcus pneumoniae Spr1875 protein fragments identified using a phage displayed genomic library.[J].2012,7(5).
- [17]Esteve-Gassent MD,Nielsen ME,Amaro C.The kinetics of antibody production in mucus and serum of European eel (Anguilla anguilla L.) after vaccination against Vibrio vulnificus: development of a new method for antibody quantification in skin mucus[J].2003,15(1).
- [18]Chung, D.-H.,Kassim, N.,Mtenga, A.B.,etc.The in vitro and in vivo efficacy of hen IgY against Vibrio parahaemolyticus and Vibrio vulnificus[J].2012,22(10).
- [19]Xiaodong Xiao,Zhongyu Zhu,Dankmeyer, Jennifer L.,etc.Human Anti-Plague Monoclonal Antibodies Protect Mice from Yersinia pestis in a Bubonic Plague Model.[J].2010,5(10).
- [20]S-J, Lee,Y H, Jung,S Y, Oh,etc.Vibrio vulnificus VvhA induces NF-κB-dependent mitochondrial cell death via lipid raft-mediated ROS production in intestinal epithelial cells.[J].2015,6.