Document Detail


A phage display approach for rapid antibody humanization: designed combinatorial V gene libraries.
MedLine Citation:
PMID:  9671778     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The development of a new strategy for antibody humanization is described. This strategy incorporates key recognition sequences from the parental rodent antibody into a phage display-based selection strategy. The original sequences of the third complementarity-determining regions (CDRs) of heavy and light chains, HCDR3 and LCDR3, were maintained and all other sequences were replaced by human sequences selected from phage-displayed antibody libraries. This approach was applied to the humanization of mouse mAb LM609 that is directed to human integrin alphav beta3 and has potential applicability in cancer therapy as an antiangiogenic agent. We demonstrate this approach (i) provides a rapid route for antibody humanization constraining the content of original mouse sequences in the final antibodies to the most hypervariable of the CDRs; (ii) generates several humanized versions with different sequences at the same time; (iii) results in affinities as high as or higher than the affinity of the original antibody; and (iv) retains the antigen and epitope specificity of the original antibody. The production of multiple humanized variants may present advantages in the selection of antibodies that are more readily expressed on a large scale and could be important in therapeutic regimens that call for long-term treatment with antibodies in which antiidiotypic responses might be avoided by administration of alternative antibodies.
Authors:
C Rader; D A Cheresh; C F Barbas
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  95     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1998 Jul 
Date Detail:
Created Date:  1998-08-20     Completed Date:  1998-08-20     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  8910-5     Citation Subset:  IM    
Affiliation:
Skaggs Institute for Chemical Biology and Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Antibodies / genetics*,  immunology
Base Sequence
Binding Sites, Antibody
Cell Line
Cloning, Molecular
DNA Primers
DNA, Complementary
Gene Library*
Humans
Immunoglobulin Variable Region / genetics
Mice
Molecular Sequence Data
Sequence Homology, Amino Acid
Grant Support
ID/Acronym/Agency:
AI 37470/AI/NIAID NIH HHS; AI 41944/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies; 0/Binding Sites, Antibody; 0/DNA Primers; 0/DNA, Complementary; 0/Immunoglobulin Variable Region
Comments/Corrections

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