S. Elliott et al., FINE-STRUCTURE EPITOPE MAPPING OF ANTIERYTHROPOIETIN MONOCLONAL-ANTIBODIES REVEALS A MODEL OF RECOMBINANT-HUMAN-ERYTHROPOIETIN STRUCTURE, Blood, 87(7), 1996, pp. 2702-2713
We have isolated and mapped the rHuEPO epitopes for three noncompeting
anti-EPO monoclonal antibodies (MoAbs). The MoAb 9G8A recognizes a li
near epitope that includes amino acids 13, 16, and 17. MoAb F12 recogn
izes a conformational epitope that includes amino acids 31 through 33,
86 through 91, and 138. MoAb D11 recognizes a conformational epitope
that includes amino acids 64 through 78 and 99 through 110. MoAb D11 n
eutralizes rHuEPO activity which suggests that its epitope may contain
the receptor binding domain. Analysis of the effect of mutations on f
olding allowed the identification of buried residues, alpha-helical, a
nd non alpha-helical regions. This data along with epitope mapping dat
a of anti rHuEPO monoclonals was used to model rHuEPO protein structur
e. A model consistent with the data is a 4-helix bundle with short and
long interconnecting loops. (C) 1996 by The American Society of Hemat
ology.