Expression, purification, and in vivo administration of a promising anti-idiotypic HIV-1 vaccine.
To date only a few neutralizing antibodies against HIV-1 exist. Since these neutralizing antibodies are only rarely found in sera of HIV-1 infected individuals an active vaccine is required. We recently developed murine anti-idiotypic antibody Ab2/3H6 against monoclonal antibody (mAb) 2F5, which is one of the most prominent neutralizing antibodies. Anti-idiotypic antibody Ab2/3H6 has been partially humanized and expressed as whole immunoglobulin G in Chinese hamster ovary cells in order to minimize the human anti-mouse antibody response. Here we describe the expression, purification, and immunohistochemical characterization of the chimeric Ab2/3H6 Fab fragment, which was finally used beside the whole IgG1 as an antigen for immunization of guinea pigs. The crude sera were screened for specific antibodies against the epitope of mAb 2F5 ELDKWA as well as for reactivity against HIV-1 gp41.
Gach
Johannes Simon
JS
Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Muthgasse 18, A-1190 Vienna, Austria. johannes.gach@boku.ac.at
Quendler
Heribert
H
Ferko
Boris
B
Katinger
Hermann
H
Kunert
Renate
R
D016915:AIDS Vaccines / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology; D000918:Antibody Specificity; D016466:CHO Cells; D002850:Chromatography, Gel; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D006168:Guinea Pigs; D015483:HIV Antibodies / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology; D015497:HIV-1 / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology; D014415:Tunicamycin / Q000494:pharmacology
D016915:AIDS Vaccines; D000888:Antibodies, Anti-Idiotypic; D015483:HIV Antibodies; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D014415:Tunicamycin