Patent ID: 8535890

Claim:
A method for determining the immunogenicity of a variant humanized CC49 antibody, comprising: contacting anti-idiotypic antibodies which bind a parental humanized CC49 antibody with the parental humanized CC49 antibody and with the variant humanized CC49 antibody; and determining the ability of the anti-idiotypic antibodies to bind the variant humanized CC49 antibody and the parental humanized CC49 antibody; wherein an at least five-fold higher molar concentration of the variant humanized CC49 antibody than the molar concentration of the parental humanized antibody being required to elicit 50% inhibition of the parental humanized CC49 antibody binding to the anti-idiotypic antibodies indicates that the variant humanized CC49 is minimally immunogenic, and wherein the parental humanized CC49 antibody comprises one of: (a) four variable light framework regions and four variable heavy framework regions of a human antibody; a light chain complementarity determining region (L-CDR)1, a L-CDR2, a L-CDR3, a heavy chain complementarity determining region (H-CDR)1, a H-CDR2, and a H-CDR3; a non-conservative substitution of a first residue at position 91, wherein the first residue is in the L-CDR3 of the antibody; and a substitution of a second residue at position 27b, wherein the second residue is in the L-CDR1 of the antibody; wherein the non-conservative substitution of the first residue at position 91 is a tyrosine to proline substitution, the substitution of the second residue at position 27b is a valine to leucine substitution, the L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3 are a HuCC49V10 antibody L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2, and H-CDR3, respectively, and the parental humanized CC49 antibody has a high binding affinity for TAG-72 and is minimally immunogenic, as compared to the HuCC49V10 antibody, deposited as ATCC Accession No. PTA-5416; (b) a light chain complementarity determining region (L-CDR)1, a L-CDR2, and a L-CDR3, a heavy chain complementarity determining region (H-CDR)1, a H-CDR2, and a H-CDR3, all from the HuCC49V10 antibody; wherein the L-CDR3 of the parental humanized CC49 antibody comprises a non-conservative amino acid substitution at position 91 and has a high binding affinity for TAG-72, as compared to the HuCC49V10 antibody, deposited as ATCC Accession No. PTA-5416; (c) humanized CC49 antibody deposited as ATCC Accession number PTA-4182; or (d) humanized CC49 antibody deposited as ATCC Accession number PTA-4183.