Biological activities on T lymphocytes of a baculovirus-expressed chimeric recombinant IgG1 antibody with specificity for the CDR3-like loop on the D1 domain of the CD4 molecule

Recombinant Anti-CD4 Antibody 13B8.2 Blocks Membrane-Proximal Events by Excluding the Zap70 Molecule and Downstream Targets SLP-76, PLCγ1, and Vav-1 from the CD4-Segregated Brij 98 Detergent-Resistant Raft Domains

“…Anti-CDR3-like rIgG 1 and Fab (rFab) 13B8.2 Abs were expressed in the baculovirus/insect cell system and purified from culture supernatant by protein A and G immunoaffinity chromatography, respectively, as previously described (17,18,26). Other anti-CD4 mAbs used were directed against the CDR2-like loop (ST4; Sanofi-Aventis), the CDR3-like loop (ST40; Sanofi-Aventis), the D2 domain (BF5; Diaclone), and the D4 domain (OKT4; Ortho Biotech).…”

“…Biological effects (6) and, above all, different abilities to relocalize CD20 in DRM (16) are also shown to be related to epitope recognition of anti-CD20 Abs. We evaluated a recombinant Ab (17,18) derived from the 13B8.2 murine anti-CD4 mAb, directed against the CDR3-like loop of CD4; this loop has not been fully exploited as a target by clinical Abs, mainly directed against other CD4 regions (11-13, 19, 20). This baculovirus-expressed recombinant Ab (rIgG 1 ) inhibits HIV replication at a post-gp120-binding step (17,18,21) and induces complement-mediated lysis, Ab-dependent cell cytotoxicity, and growth arrest of T lymphoma cells (22).…”

“…We evaluated a recombinant Ab (17,18) derived from the 13B8.2 murine anti-CD4 mAb, directed against the CDR3-like loop of CD4; this loop has not been fully exploited as a target by clinical Abs, mainly directed against other CD4 regions (11-13, 19, 20). This baculovirus-expressed recombinant Ab (rIgG 1 ) inhibits HIV replication at a post-gp120-binding step (17,18,21) and induces complement-mediated lysis, Ab-dependent cell cytotoxicity, and growth arrest of T lymphoma cells (22). The biological effects of rIgG 1 13B8.2 are partly due to signals that prevent NF-B nuclear translocation (23); ERK activation (24); and NF-AT, NF-B, and AP-1 binding to the IL-2 gene promoter (25).…”

Lepidopteran cells, an alternative for the production of recombinant antibodies?

“…Such antibodies have been shown to mediate efficient ADCC in vitro, as expected since Gal, GlcNAc and sialic acid do not play a major role in binding to FcγRIII. 23,27,28,31 Future work will no doubt see the engineering of insect cells for the production of afucosylated antibodies. Interestingly, different glycosylation patterns of recombinant antibodies have been reported in different insect tissues, such as lack of fucose reported for antibodies produced by B. mori silk gland.…”

“…Several immunoglobulin isotypes from many different species, including murine IgG, 2,3,[21][22][23][24][25] chimeric IgG, [26][27][28][29][30] human IgG, [31][32][33][34][35][36][37][38][39] IgA, [40][41][42] IgM 41 and IgE, [43][44][45] as well as pig Igs 30,45 have been successfully expressed in insect cells. These molecules are all correctly processed (signal peptide cleaved), glycosylated and assembled (H2L2) with a production rate varying from 0.75 to 100 mg/l, depending on the antibody ( …”