[Preparation and preliminary application of monoclonal antibody against recombinant human erythropoietin].
OBJECTIVE
To prepare monoclonal antibody (mAb) against erythropoietin(EPO), characterize its biological properties and use it to purify the rhEPO from transgenic goat milk.


METHODS
A crude rhEPO product was used as the antigen to immunize BALB/c mice for preparing mAbs against rhEPO. The mAbs were characterized by Western blot and indirect ELISA. Purified mAb 2E6 was coupled with pre-activated Sepharose 4B to prepare the immunoaffinity chromatography column for purifying the rhEPO from transgenic goat milk.


RESULTS
Two hybridoma cell lines(1E7 and 2E6)were obtained. mAbs 1E7 and 2E6 were shown to be IgG1 and IgG2b respectively, and their light chains were both kappa. Western blot analysis confirmed that the two mAbs could bind to rhEPO. The immunoaffinity chromatography column could adsorb 70% of rhEPO in purifying the rhEPO from transgenic goat milk.


CONCLUSIONS
Two hybridoma cell lines secreting anti-rhEPO mAbs were successfully established. The mAb-immunoaffinity chromatography column could be used to purify the rhEPO from transgenic goat milk.
Wang
Yan-fang
YF
College of Life Science and Technology, Tongji University, Shanghai 200092, China. yanfangw976@sohu.com
Peng
Jin
J
Cai
Yin-feng
YF
Ni
Ya-ming
YM
Zhao
Jian-yang
JY
Cheng
Guo-xiang
GX
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004921:Erythropoietin / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006041:Goats; D006825:Hybridomas / Q000276:immunology*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins; D004921:Erythropoietin