Open AccessCharacterization and large production of human monoclonal antibodies against the HIV‐1 envelope
Author(s) -
BOYER V.,
BROLY H.,
SOUCHE S.,
MADAULE P.,
ROSSIER J.,
ZAGURY D.,
DESGRANGES C.
Publication year - 1991
Publication title -
clinical & experimental immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 135
eISSN - 1365-2249
pISSN - 0009-9104
DOI - 10.1111/j.1365-2249.1991.tb05660.x
Subject(s) - monoclonal antibody , epitope , virology , biology , antibody , gp41 , microbiology and biotechnology , virus , vaccinia , recombinant dna , cell fusion , viral envelope , poxviridae , cell culture , immunology , gene , genetics
SUMMARY Peripheral blood lymphocytes from a volunteer immunized with a recombinant vaccinia virus VSC‐25 expressing the gp160 env protein of HTLV‐IIIB strain and from an asymptomatic HIV‐infected individual were immortalized by Epstein‐Barr (EBV). Clones which secrete human monoclonal antibodies from the two individuals (DZ, IgGl, λ and C31, IgG1, κ) were obtained and were stable for more than 2 years. The two monoclonals were directed against the gp 160 env protein of HIV, DZ directed against the gp41 and C31 directed against the gp120. C31 was group‐specific, whereas DZ was directed against the HTLV‐IIIB and HTLV‐RF strains. The epitope recognized by DZ was mapped to the carboxy terminus of the gp41, by expression of HIV DNA fragments in a yeast system and peptide analysis. The C31 epitope was not expressed by the yeast library and not present among the peptides which were tested. Monoclonal antibodies had no inhibitory effect in an HIV‐induced cell fusion assay, but DZ showed a weak neutralizing activity against the HTLV‐IIIB strain. Cloned EBV‐transformed cell lines were fused to a murine myeloma, which allowed the heteromyeloma to be cultivated in serum‐free medium. The monoclonal antibodies were produced in large quantity in a hollow‐fibre reactor at defined culture conditions and purification procedures.