Systematic exploration of the antigen binding activity of synthetic peptides isolated from the variable regions of immunoglobulins.
Sets of short (12 residues) cellulose-bound synthetic overlapping peptides derived from the sequences of the variable regions of the heavy and light chains of three different antibodies (an anti-thyroglobulin antibody, the HyHEL-5 anti-lysozyme antibody, and an anti-angiotensin II antibody) were used to systematically assess the antigen binding capacity of peptides from the antibody paratope outside their natural molecular context. Peptides enclosing one or several of the complementarity determining region (CDR) residues had antigen binding activity, although the most active peptides were not necessarily those bearing the greatest number of CDR residues. Several residues from the framework region, preceding or following the CDR, were found to play a role in binding. Affinity constants from 4.1 x 10(-7) to 6.7 x 10(-8) M-1 for the soluble form of 9 lysozyme-binding dodecapeptides were measured by BIAcore analysis. Alanine scanning of lysozyme-binding hexapeptides from the HyHEL-5 sequence identified 38 residues important for binding, of which 22 corresponded to residues that had been shown by x-ray crystallography to be at the interface between HyHEL-5 and lysozyme. Our results could be of interest for the rational identification of biologically active peptides derived from antibody sequences and in providing an experimental basis for mutagenesis of the antibody paratope.
Laune
D
D
CNRS UMR 9921. Faculté de Pharmacie, Avenue Charles Flahault, 34000 Montpellier, France.
Molina
F
F
Ferrieres
G
G
Mani
J C
JC
Cohen
P
P
Simon
D
D
Bernardi
T
T
Piechaczyk
M
M
Pau
B
B
Granier
C
C
D000595:Amino Acid Sequence; D000804:Angiotensin II / Q000276:immunology; D000818:Animals; D000941:Antigens / Q000378:metabolism*; D015374:Biosensing Techniques; D002645:Chickens; D018604:Epitope Mapping / Q000379:methods*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009113:Muramidase / Q000276:immunology; D010455:Peptides / Q000138:chemical synthesis*; D013954:Thyroglobulin / Q000276:immunology
D000941:Antigens; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D010455:Peptides; D000804:Angiotensin II; D013954:Thyroglobulin; D009113:Muramidase