Development of Enzyme Linked Immunosorbent Assay for Erythropoietin.
10.3343/kjlm.2006.26.3.185
- Author:
Ki Hong KIM
1
;
Jung Hyun SHIM
;
Min Chul CHO
;
Jeong Woo KANG
;
Hyo Eun YOON
;
Do Young YOON
;
Jong Wan KIM
;
Dong Ju SON
;
Jae Woong LEE
;
Eun Sook JEONG
;
Jin Tae HONG
;
Dong Cheul MOON
Author Information
1. Department of Bioscience and Biotechology, Konkuk University, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
EPO;
ELISA;
HRP;
Biotinylated Ab;
Streptavidin-HRP
- MeSH:
Antibodies;
Enzyme-Linked Immunosorbent Assay*;
Erythropoietin*
- From:The Korean Journal of Laboratory Medicine
2006;26(3):185-191
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: The aim of our study was to optimize and establish erythropoietin (EPO) enzyme linked immunosorbent assay (ELISA) system. METHODS: We prepared several monoclonal and polyclonal antibodies specific to human-EPO. The best combinations of antibodies for coating and detecting antibodies were selected for the establishment of ELISA. We tested several methods such as a competitive EIA and a sandwich ELISA. RESULTS: The best sandwich ELISA was optimized compared to competitive EIA when purified polyclonal antibody (PoAb) was used as a coating antibody and biotinylated PoAb as a detecting antibody. This sandwich ELISA easily detected EPO when PoAb pairs were used compared to the ELISA using monoclonal antibody and PoAb. There were no significant differences between the effects of various blocking solutions on the performance of sandwich ELISA using biotinylated antibody. The ELISA system using PBST containing 3% BSA as a blocking solution can sensitively detect EPO (10 mU/mL) in a broad range of EPO concentrations (10-2,000 mU/mL) and there were cross-reactions with other cytokines). CONCLUSIONS: EPO can be easily determined by using biotinylated PoAb as a detecting antibody and another PoAb as a coating antibody.
