title: The production, characterisation and application of monoclonal antibodies generated by immunisation with HIV-1C clade RGP140 envelope protein.
abstract: The native HIV-1 envelope spike exists as trimers on the virion surface. Therefore antibodies elicited following immunisation with trimeric envelope vaccines may be directed against epitopes found on functional envelope spikes. Novel monoclonal antibodies were produced by priming mice with plasmid DNA expressing either HIV-1 CN54 or ZM96 gp140 sequences, and boosting with CN54 trimeric rgp140. Spleen cells were fused with NS-0 cells and hybridomas were screened for production of antibodies which bound trimeric rgp140. 18 monoclonal antibodies (MAbs) were isolated from 16 colonies, which were characterised by binding in ELISA and Western blotting, neutralisation of pseudotyped viruses and isotype. Specificity for the third variable (V3) region was detected in 5 of these antibodies, suggesting that the trimeric protein did not alter the main focus of the response compared with monomeric protein. 3 MAbs were identified as neutralising in a pseudotype assay, all of which were mapped to the V3 region. Although cross clade binding of antibodies was detected the neutralisation was C clade specific.
authors: Division of Virology, National Institute for Biological Standards and Control, Blanche Lane, Potters Bar, Herts EN6 3QG, UK.
authors: Mark
authors: M
authors: Hassall
authors: Mark
authors: M
authors: Page
authors: Mark
authors: M
authors: Robinson
authors: Simon
authors: S
authors: Jeffs
authors: Ian
authors: I
authors: Jones
authors: Hongying
authors: H
authors: Chen
authors: Michael S
authors: MS
authors: Seaman
authors: Debbie
authors: D
authors: Ferguson
authors: Neil
authors: N
authors: Almond
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D015483:HIV Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D051379:Mice; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D011994:Recombinant Proteins; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1
keywords: 4-chloro-1-naphthol/diaminobenzidine tetrahydrochloride; CHO; CN/DAB; Chinese hamster ovary; Clade C; HIV-1; Mab; Monoclonal antibody; Neutralisation; TMB/DMSO; Trimer; V3; gp140; monoclonal antibody; tetramethylbenzidine/dimethylsulphoxide; third variable region