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Engineered antibody-drug conjugates with defined sites and stoichiometries of drug attachment.
McDonagh, Charlotte F; Turcott, Eileen; Westendorf, Lori; Webster, Jennifer B; Alley, Stephen C; Kim, Kristine; Andreyka, Jamie; Stone, Ivan; Hamblett, Kevin J; Francisco, Joseph A; Carter, Paul.
Afiliación
  • McDonagh CF; Seattle Genetics, Inc. 21823 30th Drive SE, Bothell, WA 98021, USA. cmcdonagh@seagen.com
Protein Eng Des Sel ; 19(7): 299-307, 2006 Jul.
Article en En | MEDLINE | ID: mdl-16644914
The chimeric anti-CD30 IgG1, cAC10, conjugated to eight equivalents of monomethyl auristatin E (MMAE) was previously shown to have potent antitumor activity against CD30-expressing tumors xenografts in mice. Moreover, the therapeutic index was increased by lowering the stoichiometry from 8 drugs/antibody down to 2 or 4. Limitations of such 'partially-loaded' conjugates are low yield (10-30%) as they are purified from mixtures with variable stoichiometry (0-8 drugs/antibody), and heterogeneity as the 2 or 4 drugs are distributed over eight possible cysteine conjugation sites. Here, the solvent-accessible cysteines that form the interchain disulfide bonds in cAC10 were replaced with serine, to reduce the eight potential conjugation sites down to 4 or 2. These Cys-->Ser antibody variants were conjugated to MMAE in near quantitative yield (89-96%) with defined stoichiometries (2 or 4 drugs/antibody) and sites of drug attachment. The engineered antibody-drug conjugates have comparable antigen-binding affinities and in vitro cytotoxic activities with corresponding purified parental antibody-drug conjugates. Additionally, the engineered and parental antibody-drug conjugates have similar in vivo properties including antitumor activity, pharmacokinetics and maximum tolerated dose. Our strategy for generating antibody-drug conjugates with defined sites and stoichiometries of drug loading is potentially broadly applicable to other antibodies as it involves engineering of constant domains.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Oligopéptidos / Inmunoconjugados / Anticuerpos Monoclonales / Antineoplásicos Límite: Animals Idioma: En Revista: Protein Eng Des Sel Asunto de la revista: BIOQUIMICA / BIOTECNOLOGIA Año: 2006 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Oligopéptidos / Inmunoconjugados / Anticuerpos Monoclonales / Antineoplásicos Límite: Animals Idioma: En Revista: Protein Eng Des Sel Asunto de la revista: BIOQUIMICA / BIOTECNOLOGIA Año: 2006 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido