title: Preparation and evaluation of human-murine chimeric antibody against protective antigen of Bacillus anthracis.
abstract: The aim of this research is to develop a human/murine chimeric Fab antibody which neutralizes the anthrax toxin, protective antigen (PA). The chimeric Fab was constructed using variable regions of murine anti-PA monoclonal antibody in combination with constant regions of human IgG. The chimeric PA6-Fab was expressed in E. coli. BL21 and evaluated by ELISA and co-immunoprecipitation- mass spectra. The potency of PA6-Fab to neutralize LeTx was examined in J774A.1 cell viability in vitro and in Fisher 344 rats in vivo. The PA6-Fab did not have domain similarity corresponding to the current anti PA mAbs, but specifically bound to anthrax PA at an affinity of 1.76 nM, and was able to neutralize LeTx in vitro and protected 56.9% cells at 20 μg/mL against anthrax LeTx. One hundred μg PA6-Fab could neutralize 300 μg LeTx in vivo. The PA6-Fab has potential as a therapeutic mAb for treatment of anthrax.
authors.affiliation: Huadong Medical Institute of Biotechniques, Nanjing 210002, China. estherhaonj@163.com.
authors.forename: Lina
authors.initials: L
authors.lastname: Hao
authors.affiliation: Huadong Medical Institute of Biotechniques, Nanjing 210002, China. zhengf82@gmail.com.
authors.forename: Feng
authors.initials: F
authors.lastname: Zheng
authors.affiliation: Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China. xiongsp_2011@163.com.
authors.forename: Siping
authors.initials: S
authors.lastname: Xiong
authors.affiliation: Huadong Medical Institute of Biotechniques, Nanjing 210002, China. hudan1234@163.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Hu
authors.affiliation: Huadong Medical Institute of Biotechniques, Nanjing 210002, China. lvheng570@gmail.com.
authors.forename: Heng
authors.initials: H
authors.lastname: Lv
authors.affiliation: Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China. pippot@139.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China. yangjin27@139.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
authors.affiliation: Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, China. fengzhenqing@njmu.edu.cn.
authors.forename: Zhenqing
authors.initials: Z
authors.lastname: Feng
authors.affiliation: Huadong Medical Institute of Biotechniques, Nanjing 210002, China. science2008@hotmail.com.
authors.forename: Changjun
authors.initials: C
authors.lastname: Wang
authors.affiliation: Huadong Medical Institute of Biotechniques, Nanjing 210002, China. zhujin1968@njmu.edu.cn.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000881:Anthrax / Q000276:immunology / Q000628:therapy*; D057134:Antibodies, Neutralizing / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D000942:Antigens, Bacterial / Q000276:immunology; D001408:Bacillus anthracis / Q000276:immunology*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D007167:Immunotherapy; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D011916:Rats, Inbred F344; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*
chemical_list: D057134:Antibodies, Neutralizing; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins; C030325:anthrax toxin