title: [Preparation and characterization of monoclonal antibody against gp120 V1/V2 domain of HIV-1 subtype CRF01_AE].
abstract: Objective To express the V1/V2 domain of HIV-1 subtype CRF01_AE gp120 in eukaryotic cells, and then prepare its monoclonal antibody (mAb) and identify its antigen reactivity. Methods Eukaryotic expression vector of pTriEx-3-V1/V2CNE55 was constructed and transiently transfected into HEK293F cell line. The V1/V2-His chimera protein was purified and injected into BALB/c mice. After the fusion between spleen cells of immunized BALB/c mice and myeloma cells SP 2/0, ELISA was used for screening the positive hybridoma cell clones against the V1/V2 recombinant protein. The specificity, titer and type of its mAb were characterized. Results We obtained a stable hybridoma cell line which secreted anti-HIV-1 AE subtype gp120 V1/V2 domain mAb. The ascite titer of the mAb was 1:81 000, and the type of the mAb was IgG1/κ. Western blotting showed that the mAb could recognize recombinant HIV-1 gp120 of different HIV-1 subtypes. Conclusion The study prepared successfully the anti-HIV-1 V1/V2 domain mAb.
authors.affiliation: Central Laboratory, Affiliated Wujin Hospital, Jiangsu University, Changzhou 213017, China. *Corresponding authors, E-mail:chy7994@163.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chu
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Xiangdan
authors.initials: X
authors.lastname: Gong
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Gao
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Airong
authors.initials: A
authors.lastname: Su
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Deyan
authors.initials: D
authors.lastname: Chen
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Hongyong
authors.initials: H
authors.lastname: Song
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China.
authors.forename: Xilin
authors.initials: X
authors.lastname: Wu
authors.affiliation: Center for Public Health Research, Medical School, Nanjing University, Nanjing 210093, China. *Corresponding authors, E-mail: wzhw@nju.edu.cn.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Wu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D005260:Female; D015483:HIV Antibodies / Q000032:analysis / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015658:HIV Infections / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D000072417:Protein Domains
chemical_list: D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120