Identification of distinct antibody epitopes and mimotopes from a peptide array of 5520 randomly generated sequences.
We used a relatively small library of 5520 randomly generated single 15-mer peptides prepared by SPOT synthesis as an array of 28.5x19.0 cm to identify epitopes for three distinct monoclonal antibodies, namely anti-p24 (human immunodeficiency virus (HIV)-1) monoclonal anibody (mab) CB4-1, anti-interleukin-10 (IL-10) mab CB/RS/13, and anti-transforming growth factor alpha (TGFalpha) mab Tab2. Initially identified peptide ligands mostly had very low affinities for the antibodies with dissociation constants around 10(-4) M. Subsequent identification of residues critical for the antibody interactions involved complete L-amino acid substitutional analyses. Several substitutions resulted in analogs with dissociation constants in the low micromolar and high nanomolar range. Specifically binding peptides with key residue patterns matching the wild-type epitopes were identified for all three antibodies. In addition, for antibody CB4-1 mimotopes that showed no homology to the known epitope were selected. Our results suggest that a very limited library diversity, although far from covering the entire sequence repertoire, can suffice to rapidly and economically select peptidic antibody epitopes and mimotopes.
Reineke
Ulrich
U
Jerini AG, Invalidenstr. 130, D-10115 Berlin, Germany. reineke@jerini.de
Ivascu
Claudia
C
Schlief
Marén
M
Landgraf
Christiane
C
Gericke
Seike
S
Zahn
Grit
G
Herzel
Hanspeter
H
Volkmer-Engert
Rudolf
R
Schneider-Mergener
Jens
J
D020650:Combinatorial Chemistry Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods* / Q000639:trends; D000939:Epitopes; D010452:Peptide Biosynthesis / Q000276:immunology*; D019151:Peptide Library
D000939:Epitopes; D019151:Peptide Library