Immune recognition of human colonic-tumour-associated MUC-2 mucins using an anti-peptide antibody.
In human intestinal malignancy, alterations occur in the expression of mucins defined by the MUC-2 gene. These changes include the unmasking of epitopes in the mucin protein core. In order to probe these modifications associated with mucins of the malignant phenotype, a monoclonal antibody (MAb) was developed against synthetic peptide with a sequence based upon that of the protein core of the MUC-2 mucin. The antibody (designated 996) was shown to recognize a high-molecular-weight glycoprotein from colonic carcinoma tissue. The material reacted uniformly with Concanavalin A but variably with other lectins, indicating heterogeneity in the associated oligosaccharide side chains. The protein core was accessible both to 996 antibody binding and to degradation with proteases. Immunization with the affinity-purified mucin-like material elicited antibodies reactive with both the immunogen and the synthetic peptides, confirming the immunogenic character of protein-core determinants. Epitope mapping studies, using synthetic peptides in solution and synthetic peptides tethered to the heads of plastic pins, indicated that the minimum epitope for the 996 antibody is a tetramer of T G T Q. Antibody interaction with the glutamine (Q) residue was determined to be of major importance in the antigen-antibody reaction. The findings illustrate the characterization of an anti-peptide antibody which may be used to probe alterations in MUC-2 mucin expression associated with human intestinal malignant disease.
Price
M R
MR
Cancer Research Laboratories, University of Nottingham, UK.
Sekowski
M
M
Ladányi
A
A
Uray
K
K
Ma
Y
Y
Durrant
L
L
Tendler
S J
SJ
D000327:Adsorption; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002918:Chymotrypsin / Q000494:pharmacology; D003110:Colonic Neoplasms / Q000276:immunology*; D003208:Concanavalin A; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005973:Glutamine / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D055262:Mucin-2; D009077:Mucins / Q000276:immunology*; D009363:Neoplasm Proteins / Q000276:immunology*; D014357:Trypsin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000939:Epitopes; C524625:MUC2 protein, human; D055262:Mucin-2; D009077:Mucins; D009363:Neoplasm Proteins; D005973:Glutamine; D003208:Concanavalin A; D002918:Chymotrypsin; D014357:Trypsin