title: Effect of core epitope modification on the antibody recognition of a MUC2 mucin peptide.
abstract: Identification of protein epitopes via combinatorial chemistry was one of the most important discoveries of the past three decades. Mapping of linear antibody epitopes can be achieved rapidly and cost-effectively by the polymer pin-bound peptide approach. In this article, the determination of the fine epitope structure of MUC2 mucin glycoprotein is described by using specific monoclonal antibody. We have used positional scanning combinatorial approach, and also parallel synthesis. The residues within the MUC2 epitope (18)PTGTQ(22) of MAb 996 were replaced by all other proteinogenic amino acids on pin-bound peptide libraries, and their antibody binding was studied in modified ELISA. Thr(19) was the least important of the residues in antibody recognition; most of the other amino acids could be replaced, except Pro. The other residues cannot be replaced without loss of antibody binding, where both the size and character of the amino acids were important. The significance of the non-chiral Gly(20) residue was further studied by competitive ELISA of parallelly synthesized soluble peptides containing L - or D-Ala instead of Gly residue. However, the D-Ala-containing oligopeptides showed no antibody binding; therefore, the backbone conformation is characteristic of that of L-amino acid containing peptides in this position as well. With the combinatorial approach we obtained relevant information about the contribution of individual amino acid side chains to the MAb 996 antibody binding within the PTGTQ predominant MUC2 mucin epitope. These results could be utilized for the design of synthetic antigens for detection of MUC2 protein core-specific antibodies related to carcinoma(s).
authors.affiliation: Research Group of Peptide Chemistry, Eötvös L. University, Hungarian Academy of Sciences, P.O.B. 32, Budapest 112, 1518, Hungary. uray@elte.hu
authors.forename: Katalin
authors.initials: K
authors.lastname: Uray
authors.forename: Ferenc
authors.initials: F
authors.lastname: Hudecz
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D006031:Glycosylation; D008969:Molecular Sequence Data; D055262:Mucin-2 / Q000737:chemistry / Q000276:immunology*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D011485:Protein Binding
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000939:Epitopes; D055262:Mucin-2; D019151:Peptide Library; D010455:Peptides