Interaction of the octapeptide angiotensin II with a high-affinity single-chain Fv and with peptides derived from the antibody paratope

P Cohen; D Laune; I Teulon; T Combes; M Pugnière; G Badouaille; C Granier; J C Mani; D Simon

doi:10.1016/s0022-1759(01)00395-7

Interaction of the octapeptide angiotensin II with a high-affinity single-chain Fv and with peptides derived from the antibody paratope

J Immunol Methods. 2001 Aug 1;254(1-2):147-60. doi: 10.1016/s0022-1759(01)00395-7.

Authors

P Cohen¹, D Laune, I Teulon, T Combes, M Pugnière, G Badouaille, C Granier, J C Mani, D Simon

Affiliation

¹ Department of Immunology/Oncology, Sanofi-Synthelabo, 371 rue du Professeur Blayac, 34184 Montpellier Cedex 4, France.

PMID: 11406160
DOI: 10.1016/s0022-1759(01)00395-7

Abstract

The amino-acid sequence of the very high-affinity anti-angiotensin II monoclonal antibody 4D8 was predicted from the nucleotide sequence of the heavy and light chain variable genes. The single-chain variable fragment (scFv) was constructed and expressed in Escherichia coli as a soluble protein and at the surface of the filamentous M13 phage and was compared with the full-length antibody (Ab). The scFv showed the same specificity profile and affinity constant as the intact antibody (5.0x10(10) and 8.0x10(10) M(-1), respectively, by Scatchard analysis). Several peptides from the set of overlapping dodecapeptides covering the variable domains of 4D8 mAb were found to specifically bind biotinylated angiotensin II: peptides from the L1, L2, L3 and H1 regions had the strongest capacity to bind the antigen.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Angiotensin II / immunology*
Antibody Affinity
Antibody Specificity
Base Sequence
Escherichia coli / metabolism
Gene Expression
Immunoglobulin Fragments / genetics
Immunoglobulin Fragments / immunology*
Immunoglobulin Fragments / isolation & purification
Immunoglobulin Heavy Chains / genetics
Immunoglobulin Heavy Chains / immunology*
Immunoglobulin Heavy Chains / isolation & purification
Immunoglobulin Light Chains / genetics
Immunoglobulin Light Chains / immunology*
Immunoglobulin Light Chains / isolation & purification
Immunoglobulin Variable Region / genetics
Immunoglobulin Variable Region / immunology*
Immunoglobulin Variable Region / isolation & purification
Molecular Sequence Data
Oligopeptides / immunology
Recombinant Proteins / genetics
Recombinant Proteins / immunology
Recombinant Proteins / isolation & purification
Sequence Alignment
Solubility

Substances

Immunoglobulin Fragments
Immunoglobulin Heavy Chains
Immunoglobulin Light Chains
Immunoglobulin Variable Region
Oligopeptides
Recombinant Proteins
immunoglobulin Fv
Angiotensin II