Preparation and preclinical evaluation of humanised A33 immunoconjugates for radioimmunotherapy Journal Article
| Authors: | King, D. J.; Antoniw, P.; Owens, R. J.; Adair, J. R.; Haines, A. M. R.; Farnsworth, A. P. H.; Finney, H.; Lawson, A. D. G.; Lyons, A.; Baker, T. S.; Baldock, D.; Mackintosh, J.; Gofton, C.; Yarranton, G. T.; McWilliams, W.; Shochat, D.; Leichner, P. K.; Welt, S.; Old, L. J.; Mountain, A. |
| Article Title: | Preparation and preclinical evaluation of humanised A33 immunoconjugates for radioimmunotherapy |
| Abstract: | A humanised IgG1/k version of A33 (hA33) has been constructed and expressed with yields up to 700 mg l-1 in mouse myeloma NS0 cells in suspension culture. The equilibrium dissociation constant of hA33 (KD = 1.3 nM) was shown to be equivalent to that of the murine antibody in a cell-binding assay. hA33 labelled with yttrium-90 using the macrocyclic chelator 12N4 (DOTA) was shown to localise very effectively to human colon tumour xenografts in nude mice, with tumour levels increasing as blood concentration fell up to 144 h. A Fab’ variant of hA33 with a single hinge thiol group to facilitate chemical cross-linking has also been constructed and expressed with yields of 500 mg l-1. Trimaleimide cross-linkers have been used to produce a trivalent Fab fragment (hA33 TFM) that binds antigen on tumour cells with greater avidity than hA33 IgG. Cross-linkers incorporating 12N4 or 9N3 macrocycles have been used to produce hA33 TFM labelled stably and site specifically with yttrium-90 or indium-111 respectively. These molecules have been used to demonstrate that hA33 TFM is cleared more rapidly than hA33 IgG from the circulation of animals but does not lead to accumulation of these metallic radionuclides in the kidney. 90Y-labelled hA33 TFM therefore appears to be the optimal form of the antibody for radioimmunotherapy of colorectal carcinoma. © 1995 Stockton Press. All rights reserved. |
| Keywords: | controlled study; human cell; nonhuman; cancer radiotherapy; binding affinity; animal cell; animals; mice; cancer immunotherapy; multiple myeloma; gene expression; tumor xenograft; drug screening assays, antitumor; tumor cells, cultured; colorectal neoplasms; cloning, molecular; indium radioisotopes; amino acid sequence; molecular sequence data; immunoglobulin g; isotope labeling; tissue distribution; mice, nude; colon tumor; tumor cell; transplantation, heterologous; neoplasm transplantation; antigen binding; radioimmunotherapy; indium 111; antibody; yttrium; yttrium radioisotopes; immunoglobulin g antibody; dna, complementary; cho cells; cricetinae; dissociation constant; immunoglobulin f(ab) fragment; cross linking; immunoglobulin fragments; antibody conjugate; immunoconjugates; yttrium 90; myeloma cell; antibody blood level; chelating agent; hybridomas; genes, immunoglobulin; plasma clearance; humans; human; priority journal; article; thiol group; tri-fab |
| Journal Title: | British Journal of Cancer |
| Volume: | 72 |
| Issue: | 6 |
| ISSN: | 0007-0920 |
| Publisher: | Nature Publishing Group |
| Date Published: | 1995-12-01 |
| Start Page: | 1364 |
| End Page: | 1372 |
| Language: | English |
| DOI: | 10.1038/bjc.1995.516 |
| PUBMED: | 8519646 |
| PROVIDER: | scopus |
| PMCID: | PMC2034099 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 28 August 2018 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work