Effect of core epitope modification on the antibody recognition of a MUC2 mucin peptide

Research output: Contribution to journalArticle

3 Citations (Scopus)

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 18PTGTQ 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).

Original languageEnglish
Pages (from-to)103-112
Number of pages10
JournalMolecular Diversity
Volume16
Issue number1
DOIs
Publication statusPublished - Feb 2012

Fingerprint

Epitopes
Mucins
antibodies
Antibodies
Peptides
peptides
Amino acids
amino acids
Amino Acids
Enzyme-Linked Immunosorbent Assay
Proteins
Peptide Library
Oligopeptides
Glycoproteins
Synthetic Vaccines
Monoclonal antibodies
proteins
antigens
Conformations
Polymers

Keywords

  • Combinatorial scanning
  • Mapping of linear antibody epitope
  • MUC2 mucin
  • Pin-bound peptides
  • Positional scanning

ASJC Scopus subject areas

  • Molecular Biology
  • Drug Discovery
  • Information Systems
  • Physical and Theoretical Chemistry
  • Inorganic Chemistry
  • Catalysis
  • Organic Chemistry

Cite this

Effect of core epitope modification on the antibody recognition of a MUC2 mucin peptide. / Uray, K.; Hudecz, F.

In: Molecular Diversity, Vol. 16, No. 1, 02.2012, p. 103-112.

Research output: Contribution to journalArticle

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