BMB Reports
- Volume 30 Issue 5
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- Pages.315-319
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- 1997
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- 1976-670X(eISSN)
Monoclonal Antibody CFC-6, which Binds to Helix II, Inhibits Erythropoietin-Induced Bioactivity
- Ha, Byung-Jhip (R & D center, Cheiljedang Co.) ;
- Kim, Suk-Joon (R & D center, Cheiljedang Co.) ;
- Park, Ji-Sook (R & D center, Cheiljedang Co.) ;
- Yoo, Ree-Ann (R & D center, Cheiljedang Co.) ;
- Lee, Dong-Eok (Korean Advanced Institute of Science and Technology) ;
- Yoo, Ook-Joon (Korean Advanced Institute of Science and Technology) ;
- Woo, Koo (Pharmaceutical Division, Cheiljedang Co.) ;
- Kim, Hyun-Su (R & D center, Cheiljedang Co.) ;
- Oh, Myung-Suk (R & D center, Cheiljedang Co.)
- Published : 1997.09.30
Abstract
It was discovered that monoclonal anti-erythropoietin (EPO) antibody CFC-6 can neutralize EPO-induced cell activation. To know the binding site of CFC-6, recombinant human erythropoietin (rhEPO) was digested with Glu-C, followed by a separation using high performance liquid chromato graphy (HPLC). Each HPLC fraction was blotted on the nitrocellulose membrane and the membrane was treated with anti-EPO antibody CFC-6 and anti-mouse antibody which is modified with peroxidase. Only one spot showed the color and the fraction was sequenced by Edman degradation. The results suggest that CFC-6 recognizes amino acid sequence V63-W-Q-G-L-A-L-L-S-E72 which is a part of helix II of the EPO molecule. Binding of CFC-6 to EPO may inhibit EPO binding to its receptor, which implies that the antibody binding site and the receptor binding site are close or overlapping.