Skip Navigation


PEDS Advance Access originally published online on April 27, 2006
Protein Engineering Design and Selection 2006 19(7):299-307; doi:10.1093/protein/gzl013
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow All Versions of this Article:
19/7/299    most recent
gzl013v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (8)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by McDonagh, C. F.
Right arrow Articles by Carter, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McDonagh, C. F.
Right arrow Articles by Carter, P.
Related Collections
Right arrow 2006
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Engineered antibody–drug conjugates with defined sites and stoichiometries of drug attachment

Charlotte F. McDonagh3, Eileen Turcott, Lori Westendorf, Jennifer B. Webster, Stephen C. Alley, Kristine Kim, Jamie Andreyka, Ivan Stone, Kevin J. Hamblett1, Joseph A. Francisco2 and Paul Carter3

Seattle Genetics, Inc. 21823 30th Drive SE, Bothell, WA 98021, USA

3To whom correspondence should be addressed. E-mail: pcarter{at}seagen.com; cmcdonagh{at}seagen.com

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.

Keywords: auristatin/CD30/conjugate/engineered antibody

Received March 7, 2006; accepted March 14, 2006.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Clin. Cancer Res.Home page
E. Oflazoglu, I. J. Stone, K. Gordon, C. G. Wood, E. A. Repasky, I. S. Grewal, C.-L. Law, and H.-P. Gerber
Potent Anticarcinoma Activity of the Humanized Anti-CD70 Antibody h1F6 Conjugated to the Tubulin Inhibitor Auristatin via an Uncleavable Linker
Clin. Cancer Res., October 1, 2008; 14(19): 6171 - 6180.
[Abstract] [Full Text] [PDF]

Home page
 Molecular Cancer TherapeuticsHome page
C. F. McDonagh, K. M. Kim, E. Turcott, L. L. Brown, L. Westendorf, T. Feist, D. Sussman, I. Stone, M. Anderson, J. Miyamoto, et al.
Engineered anti-CD70 antibody-drug conjugate with increased therapeutic index
Mol. Cancer Ther., September 1, 2008; 7(9): 2913 - 2923.
[Abstract] [Full Text] [PDF]

Home page
 Molecular Cancer TherapeuticsHome page
M. C. Ryan, M. Hering, D. Peckham, C. F. McDonagh, L. Brown, K. M. Kim, D. L. Meyer, R. F. Zabinski, I. S. Grewal, and P. J. Carter
Antibody targeting of B-cell maturation antigen on malignant plasma cells
Mol. Cancer Ther., November 1, 2007; 6(11): 3009 - 3018.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
D. P. Humphreys, S. P. Heywood, A. Henry, L. Ait-Lhadj, P. Antoniw, R. Palframan, K. J. Greenslade, B. Carrington, D. G. Reeks, L. C. Bowering, et al.
Alternative antibody Fab' fragment PEGylation strategies: combination of strong reducing agents, disruption of the interchain disulphide bond and disulphide engineering
Protein Eng. Des. Sel., May 1, 2007; 20(5): 227 - 234.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.