title: Interaction of the octapeptide angiotensin II with a high-affinity single-chain Fv and with peptides derived from the antibody paratope.
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.
authors.affiliation: Department of Immunology/Oncology, Sanofi-Synthelabo, 371 rue du Professeur Blayac, 34184 Montpellier Cedex 4, France.
authors.forename: P
authors.initials: P
authors.lastname: Cohen
authors.forename: D
authors.initials: D
authors.lastname: Laune
authors.forename: I
authors.initials: I
authors.lastname: Teulon
authors.forename: T
authors.initials: T
authors.lastname: Combes
authors.forename: M
authors.initials: M
authors.lastname: Pugnière
authors.forename: G
authors.initials: G
authors.lastname: Badouaille
authors.forename: C
authors.initials: C
authors.lastname: Granier
authors.forename: J C
authors.initials: JC
authors.lastname: Mani
authors.forename: D
authors.initials: D
authors.lastname: Simon
mesh_terms: D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D016415:Sequence Alignment; D012995:Solubility
chemical_list: D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D009842:Oligopeptides; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D000804:Angiotensin II