Definition of essential amino acid residues in the recognition of a peptide by a mouse monoclonal antibody.
A mouse monoclonal antibody reacting in ELISA with a synthetic peptide representing a linear amino acid stretch of the protein antigen was tested on all overlapping 5-mer to 9-mer fragments of the peptide, as prepared by multi-pin synthesis. Analysis of the binding data suggests that several residues in the peptide might be relatively unrelevant for recognition, while few others seem to play a critical role as key residues. On the basis of such observations, we attempted to reconstruct an alternative essential epitope by introducing multiple amino acid substitutions in the 9-mer peptide exhibiting the best binding activity, and then tested its ability to be recognized by the monoclonal antibody.
Rosanó
L
L
Istituto Regina Elena for Cancer Research, Laboratory of Biochemistry, Roma, Italy.
Di Modugno
F
F
Romagnoli
G
G
Chersi
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry*; D007074:Immunoglobulin G; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D010446:Peptide Fragments