Construction and characterization of a single-chain catalytic antibody

Richard A. Gibbs; Bruce A. Posner; David R. Filpula; Steven W. Dodd; Malcolm A J Finkelman; Timothy K. Lee; Marie Wroble; Marc Whitlow; Stephen J. Benkovic

doi:10.1073/pnas.88.9.4001

Construction and characterization of a single-chain catalytic antibody

Richard A. Gibbs, Bruce A. Posner, David R. Filpula, Steven W. Dodd, Malcolm A J Finkelman, Timothy K. Lee, Marie Wroble, Marc Whitlow, Stephen J. Benkovic

Research output: Contribution to journal › Article › peer-review

61 Scopus citations

Abstract

The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage λ. By inserting the variable regions of this Fab coding sequence into a (NH₂)-V_L-linker-V_H-(COOH) construct (where V_L and V_H represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. This protein has been expressed in Escherichia coli and exhibits the same catalytic parameters as the parent monoclonal antibody NPN43C9. Single-chain forms of catalytic antibodies may prove valuable for structural and site-directed mutagenesis studies as well as for large-scale applications of catalytic antibodies.

Original language	English (US)
Pages (from-to)	4001-4004
Number of pages	4
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	88
Issue number	9
DOIs	https://doi.org/10.1073/pnas.88.9.4001
State	Published - 1991

Keywords

Amide hydrolysis
Antibody cloning
Escherichia coli expression
Single-chain Fv
Single-chain antigen-binding protein

ASJC Scopus subject areas

General

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10.1073/pnas.88.9.4001

Cite this

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title = "Construction and characterization of a single-chain catalytic antibody",

abstract = "The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage λ. By inserting the variable regions of this Fab coding sequence into a (NH2)-VL-linker-VH-(COOH) construct (where VL and VH represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. This protein has been expressed in Escherichia coli and exhibits the same catalytic parameters as the parent monoclonal antibody NPN43C9. Single-chain forms of catalytic antibodies may prove valuable for structural and site-directed mutagenesis studies as well as for large-scale applications of catalytic antibodies.",

keywords = "Amide hydrolysis, Antibody cloning, Escherichia coli expression, Single-chain Fv, Single-chain antigen-binding protein",

author = "Gibbs, {Richard A.} and Posner, {Bruce A.} and Filpula, {David R.} and Dodd, {Steven W.} and Finkelman, {Malcolm A J} and Lee, {Timothy K.} and Marie Wroble and Marc Whitlow and Benkovic, {Stephen J.}",

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doi = "10.1073/pnas.88.9.4001",

language = "English (US)",

volume = "88",

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T1 - Construction and characterization of a single-chain catalytic antibody

AU - Gibbs, Richard A.

AU - Posner, Bruce A.

AU - Filpula, David R.

AU - Dodd, Steven W.

AU - Finkelman, Malcolm A J

AU - Lee, Timothy K.

AU - Wroble, Marie

AU - Whitlow, Marc

AU - Benkovic, Stephen J.

PY - 1991

Y1 - 1991

N2 - The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage λ. By inserting the variable regions of this Fab coding sequence into a (NH2)-VL-linker-VH-(COOH) construct (where VL and VH represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. This protein has been expressed in Escherichia coli and exhibits the same catalytic parameters as the parent monoclonal antibody NPN43C9. Single-chain forms of catalytic antibodies may prove valuable for structural and site-directed mutagenesis studies as well as for large-scale applications of catalytic antibodies.

AB - The antigen-binding (Fab) fragment of the catalytic monoclonal antibody NPN43C9 has recently been cloned by using bacteriophage λ. By inserting the variable regions of this Fab coding sequence into a (NH2)-VL-linker-VH-(COOH) construct (where VL and VH represent the heavy and light chain variable regions), we have assembled a recombinant gene encoding a catalytic single-chain antigen-binding protein. This protein has been expressed in Escherichia coli and exhibits the same catalytic parameters as the parent monoclonal antibody NPN43C9. Single-chain forms of catalytic antibodies may prove valuable for structural and site-directed mutagenesis studies as well as for large-scale applications of catalytic antibodies.

KW - Amide hydrolysis

KW - Antibody cloning

KW - Escherichia coli expression

KW - Single-chain Fv

KW - Single-chain antigen-binding protein

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JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

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ER -

Construction and characterization of a single-chain catalytic antibody

Abstract

Keywords

ASJC Scopus subject areas

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