High-level expression and characterization of a mouse-human chimeric CD4 antibody with therapeutic potential.
The use of murine anti-CD4 monoclonal antibodies (MAbs) has shown considerable promise for the treatment of allograft rejection and rheumatoid arthritis. We have constructed mouse-human anti-CD4 antibodies with the goal of increasing their clinical potential by decreasing immunogenicity and improving effector functions. The chimeric antibodies were constructed by cloning the heavy and light chain variable regions of M-T412, a murine antibody raised against the human CD4 antigen, and joining them to the human G1, G4, or kappa constant regions in mammalian expression vectors. After transfection into mouse myeloma cells, stable cell lines were isolated that secrete up to 140 micrograms/ml chimeric antibody in static culture. The chimeric antibodies were equivalent to the murine antibody in their binding characteristics and relative affinities. However, the chimeric M-T412 MAbs have enhanced activity when compared to the murine G2a MAb in mediating antibody-dependent cell-mediated cytotoxicity using human CD4+ target and effector cells.
Looney
J E
JE
Department of Molecular Biology, Centocor, Inc., Malvern, PA 19355.
Knight
D M
DM
Arevalo-Moore
M
M
Trinh
H
H
Pak
K Y
KY
Dalesandro
M R
MR
Rieber
E P
EP
Riethmuller
G
G
Daddona
P E
PE
Ghrayeb
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000627:therapeutic use; D000920:Antibody-Dependent Cell Cytotoxicity; D015704:CD4 Antigens; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005803:Genes, Immunoglobulin; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D016176:T-Lymphocyte Subsets / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D011994:Recombinant Proteins; D004247:DNA