Epitope mapping of PR81 anti-MUC1 monoclonal antibody following PEPSCAN and phage display techniques.
PR81 is an anti-MUC1 monoclonal antibody (MAb) which was generated against human MUC1 mucin that reacted with breast cancerous tissue, MUC1 positive cell line (MCF-7, BT-20, and T-4 7 D), and synthetic peptide, including the tandem repeat sequence of MUC1. Here we characterized the binding properties of PR81 against the tandem repeat of MUC1 by two different epitope mapping techniques, namely, PEPSCAN and phage display. Epitope mapping of PR81 MAb by PEPSCAN revealed a minimal consensus binding sequence, PDTRP, which is found on MUC1 peptide as the most important epitope. Using the phage display peptide library, we identified the motif PD(T/S/G)RP as an epitope and the motif AVGLSPDGSRGV as a mimotope recognized by PR81. Results of these two methods showed that the two residues, arginine and aspartic acid, have important roles in antibody binding and threonine can be substituted by either glycine or serine. These results may be of importance in tailor making antigens used in immunoassay.
Mohammadi
Mohammad
M
Tarbiat Modares University, School of Medical Sciences, Tehran, Iran.
Rasaee
Mohammad Javad
MJ
Rajabibazl
Masoumeh
M
Paknejad
Malihe
M
Zare
Mehrak
M
Mohammadzadeh
Sara
S
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry*; D000918:Antibody Specificity; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000276:immunology*; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D018396:Mucin-1; D019151:Peptide Library