Epitope mapping of the anti-human progesterone receptor monoclonal antibody, AB-52.
The monoclonal antibody AB-52 has high affinity and specificity for the two natural human progesterone receptor forms, receptor A (hPRA) and receptor B (hPRB), but it does not bind the PR of chick, mice, rats or rabbits. We have used a novel method to map its epitope. Based on a series of site-directed mutants of hPRA, together with immunoblotting, DNA gel mobility shift and antibody super shift assays, we have mapped the epitope of AB-52 to a 17 amino acid sequence lying between Val221 and Leu237 of the 933 amino acid hPRB protein. This N-terminal sequence is common to both hPRB and hPRA but is missing in chick PR and differs extensively from mouse PR. No anti-rabbit PR antibodies map to the homologous rabbit PR sequence which differs from hPR by four amino acids, suggesting that one or more of these four amino acids form a critical subset of residues in hPR that define the human specificity of AB-52. Knowledge of the AB-52 epitope is useful in structural analysis of PR, and in competition studies. Additionally, this 17 amino acid peptide whose antigenicity would not be predicted from computer analysis, should be useful for generating additional hPR specific antibodies.
Takimoto
G S
GS
Department of Medicine, University of Colorado Health Sciences Center, Denver 80262.
Tasset
D M
DM
Miller
L A
LA
Horwitz
K B
KB
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002645:Chickens; D003001:Cloning, Molecular; D000939:Epitopes / Q000032:analysis* / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011817:Rabbits; D051381:Rats; D011980:Receptors, Progesterone / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011980:Receptors, Progesterone