Molecular modeling and preclinical evaluation of the humanized NR-LU-13 antibody
| Autor: | S S, Graves, S C, Goshorn, D M, Stone, D B, Axworthy, J M, Reno, B, Bottino, S, Searle, A, Henry, J, Pedersen, A R, Rees, R T, Libby |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Mice
Inbred BALB C Recombinant Fusion Proteins Molecular Sequence Data Drug Evaluation Preclinical Immunoglobulin Variable Region Antibodies Monoclonal Mice Nude Enzyme-Linked Immunosorbent Assay CHO Cells Neoplasms Experimental Epithelial Cell Adhesion Molecule Polymerase Chain Reaction Mice Antigens Neoplasm Cricetinae Drug Design Tumor Cells Cultured Animals Humans Immunoglobulin Light Chains Amino Acid Sequence Immunoglobulin Heavy Chains Cell Adhesion Molecules |
| Zdroj: | Clinical cancer research : an official journal of the American Association for Cancer Research. 5(4) |
| ISSN: | 1078-0432 |
| Popis: | A mouse-human chimeric monoclonal antibody (chNR-LU-13), specific for the EGP40 pancarcinoma antigen, was humanized through three-dimensional molecular modeling. Humanization of the chNR-LU-13 antibody is expected to enhance its use for patients undergoing immunotherapy. On the basis of the observed amino acid sequence identity, chNR-LU-13 complementary determining regions (CDRs) of the V(L) and V(H) regions were grafted onto the human anti-DNA-associated idiotype immunoglobulin clone, R3.5H5G'CL. Ten amino acids residues within the humanized framework were back-mutated to their corresponding chNR-LU-13 sequence, because they were predicted to disrupt the canonical classification of the CDRs or were within 5 A of a CDR. Synthesis of the V(L) and V(H) regions was accomplished by recursive PCR, and the dual-chain expression vector p451.C4 was positioned under control of the CMV(P+E). We observed by competitive ELISA that the recombinant humanized NR-LU-13 (huNR-LU-13) IgG1 antibody exhibited an indistinguishable immunoreactivity profile when compared with the murine monoclonal antibody (muNR-LU-10). The huNR-LU-13 antibody was effective in mediating both antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity when assayed against either the breast carcinoma cell line, MCF-7, or the colon adenocarcinoma cell line, SW1222. Biodistribution studies using i.v. coinjected 131I-muNR-LU-10 and 125I-huNR-LU-13 confirmed that the huNR-LU-13 specifically targets to the tumor in athymic BALB/c mice bearing the SW1222 human tumor xenograft. Humanization of the chNR-LU-13 antibody is expected to eliminate an undesired human antimouse antibody response, allowing for repeated i.v. administration into humans. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|