Recognition of peptidyl epitopes by polymorphic epithelial mucin (PEM)-specific monoclonal antibodies.
Peptidyl epitope recognition by several murine monoclonal antibodies (MAbs E29, H23, HMFG-1, HMFG-2, MA5, MA6 and MA9) which react with the polymorphic epithelial mucins [PEM; epithelial membrane antigen (EMA)] was studied by using ten synthetic peptides representative of the 20 residue tandem repeat as test antigens. Antibody binding to 6-10 residue overlaps and to peptides having a common carboxy-terminus and staggered amino-termini (8-31 residues) was assessed by solid phase and competition ELISA techniques. From these analyses, all MAbs except MA9 were found to react predominantly with the carboxy-terminal half of the repeat motif. Polyclonal antibody responses in mice immunized with intact EMA/PEM-containing preparations also displayed significant reactivities against synthetic repeat peptide antigens and, conversely, synthetic peptides as carrier-conjugated immunogens induced antibodies recognizing intact antigens. These results are discussed vis-à-vis peptide conformation, the potential effects of O-glycosylation on secondary structure, and the possible effects of these parameters on immunogenicity and antigenicity.
Dion
A S
AS
Institute of Molecular Genetics, Center for Molecular Medicine and Immunology, Newark, NJ 07103.
Smorodinsky
N I
NI
Williams
C J
CJ
Wreschner
D H
DH
Major
P P
PP
Keydar
I
I
D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000276:immunology*; D001943:Breast Neoplasms / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology*; D008895:Milk, Human / Q000276:immunology; D008969:Molecular Sequence Data; D018396:Mucin-1; D009077:Mucins / Q000276:immunology*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D012091:Repetitive Sequences, Nucleic Acid
D000951:Antigens, Neoplasm; D008562:Membrane Glycoproteins; D018396:Mucin-1; D009077:Mucins; D010446:Peptide Fragments