Document Detail


Construction, bacterial expression and characterization of a bifunctional single-chain antibody-phosphatase fusion protein targeted to the human erbB-2 receptor.
MedLine Citation:
PMID:  1369487     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have constructed genes expressing single-chain antigen binding proteins (scFv) which recognize the human erbB-2 receptor. These genes encode the heavy and light chain variable domains of an erbB-2 receptor specific monoclonal antibody, MAb FRP5, connected by a peptide linker. In order to express a bifunctional molecule, a bacterial alkaline phosphatase gene was fused 3' to the scFv gene. The scFv(FRP5) and scFv(FRP5)-alkaline phosphatase fusion protein (scFv(FRP5)-PhoA) expressed in E. coli specifically recognize the human erbB-2 protein and compete with MAb FRP5 for binding to the receptor. The bound scFv(FRP5)-PhoA protein can be detected directly on tumor cells using a substrate for alkaline phosphatase, showing that the chimeric protein retains both binding and enzymatic activity.
Authors:
W Wels; I M Harwerth; M Zwickl; N Hardman; B Groner; N E Hynes
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Bio/technology (Nature Publishing Company)     Volume:  10     ISSN:  0733-222X     ISO Abbreviation:  Biotechnology (N.Y.)     Publication Date:  1992 Oct 
Date Detail:
Created Date:  1992-11-06     Completed Date:  1992-11-06     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  8309273     Medline TA:  Biotechnology (N Y)     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1128-32     Citation Subset:  B    
Affiliation:
Friedrich Miescher Institute, Basel, Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / genetics,  metabolism*
Amino Acid Sequence
Antibodies, Monoclonal / metabolism*
Binding, Competitive
Cloning, Molecular*
Escherichia coli / genetics
Female
Genes, Bacterial
Humans
Immunohistochemistry
Molecular Sequence Data
Plasmids / chemistry*,  genetics
Proto-Oncogene Proteins / metabolism*
Receptor, erbB-2
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Proto-Oncogene Proteins; EC 2.7.10.1/Receptor, erbB-2; EC 3.1.3.1/Alkaline Phosphatase

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