1. Selective formation of covalent protein heterodimers with an unnatural amino acid.
- Author
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Hutchins BM, Kazane SA, Staflin K, Forsyth JS, Felding-Habermann B, Smider VV, and Schultz PG
- Subjects
- Antibodies, Monoclonal genetics, Antibodies, Monoclonal metabolism, Antibodies, Monoclonal, Humanized, Antineoplastic Agents, Phytogenic immunology, Antineoplastic Agents, Phytogenic toxicity, Cell Line, Tumor, Dimerization, Humans, Immunoglobulin Fab Fragments chemistry, Immunoglobulin Fab Fragments genetics, Immunoglobulin Fab Fragments metabolism, Immunotoxins immunology, Immunotoxins toxicity, Maleimides chemistry, Phenylalanine chemistry, Receptor, ErbB-2 immunology, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Ribosome Inactivating Proteins, Type 1 genetics, Ribosome Inactivating Proteins, Type 1 metabolism, Saporins, Trastuzumab, Antibodies, Monoclonal chemistry, Antineoplastic Agents, Phytogenic chemistry, Immunotoxins chemistry, Phenylalanine analogs & derivatives, Ribosome Inactivating Proteins, Type 1 chemistry
- Abstract
We report a strategy for the generation of heterodimeric protein conjugates using an unnatural amino acid with orthogonal reactivity. This paper addresses the challenges of site-specificity and homogeneity with respect to the synthesis of bivalent proteins and antibody-drug conjugates. There are numerous antibody-drug conjugates in preclinical and clinical development, yet these are based either on nonspecific lysine coupling chemistry or on disulfide modification made difficult by the large number of cysteines in antibodies. Here, we describe a recombinant approach that can be used to rapidly generate a variety of constructs with defined conjugation sites. Moreover, this methodology results in homogeneous antibody conjugates whose biological, physical, and pharmacological properties can be quantitatively assessed and subsequently optimized. As proof of concept, we have generated anti-Her2 Fab-Saporin conjugates that demonstrate excellent potency in vitro., (Copyright © 2011 Elsevier Ltd. All rights reserved.)
- Published
- 2011
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