Cloning and expression of a chimeric antibody directed against the human transferrin receptor.
The cloning, construction and expression of chimeric Ig genes, encoding a mAb directed against the human transferrin receptor, is described. From a mouse hybridoma cell line, secreting an antitransferrin receptor antibody, mRNA was prepared and converted into cDNA using Ig-specific oligonucleotides. H and L chain encoding cDNA fragments were isolated and sequenced. Chimeric genes were constructed by linking the murine V region cDNA fragments to human C region exons. After sequential transfection of nonproducing mouse hybridoma cells with the expression vectors containing the chimeric H and L chain genes, antibody secreting transfectomas were obtained. ELISA and immunoblot analysis clearly demonstrate the secretion of human kappa- and gamma-1 chain. Flow microfluorimetry analysis of the chimeric antibody shows that the Ag-binding capacity has been retained. The chimeric antibody most likely will be less immunogenic then the original mouse antibody when used in human cancer therapy.
Hoogenboom
H R
HR
Dr. L. Willems Instituut, Limburgs Universitair Centrum, Diepenbeek, Belgium.
Raus
J C
JC
Volckaert
G
G
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005803:Genes, Immunoglobulin; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011233:Precipitin Tests; D011990:Receptors, Transferrin / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D004247:DNA