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Application of a bridging ELISA for detection of anti-erythropoietin binding antibodies and a cell-based bioassay for neutralizing antibodies in human sera

Authors
 Sug Kyun Shin  ;  Sung Kyu Ha  ;  Kang Wook Lee  ;  Tae-Hyun Yoo  ;  Sung-Ro Yun  ;  Se-Hee Yoon  ;  Sung-Jo Kim  ;  Soo-Kyeong Lee  ;  Tae-Hwe Heo 
Citation
 JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, Vol.52(2) : 289-293, 2010 
Journal Title
 JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 
ISSN
 0731-7085 
Issue Date
2010
MeSH
Antibodies, Anti-Idiotypic/immunology* ; Antibodies, Neutralizing/immunology* ; Biological Assay/methods ; Enzyme-Linked Immunosorbent Assay/methods* ; Erythropoietin/immunology* ; Humans ; Recombinant Proteins ; Serum/immunology*
Keywords
Erythropoietin ; Pure red-cell aplasia ; Bridging ELISA ; Neutralizing bioassay ; Immunogenicity
Abstract
Although erythropoietin (EPO)-related pure red-cell aplasia (PRCA) is a rare disorder, attention still needs to be paid because underline mechanism of EPO immunogenicity is various and controversial. Among several assay systems for screening of anti-EPO binding antibodies (Abs), we adopted and setup the bridging ELISA using streptavidin-coated plate. To test their neutralizing activities, cell-based neutralizing (NT) bioassay was setup. When we analyzed serum samples by using these two assays, we found two positive results in the two samples. In the sample 1, 411.9 ng/ml of anti-EPO Abs were found and neutralizing activity of 36.2% at 1:5 serum dilution was detected. In the sample 2, 40.5 ng/ml of anti-EPO Abs were found and neutralizing activity of 96.7% was detected. Our results indicate that the higher anti-EPO antibody (Ab) level in a serum does not always lead to the stronger neutralizing activity. This report gives crucial consideration to the needs of establishing clear criteria to link various assay parameters with the onset of PRCA and its progression.
Full Text
http://www.sciencedirect.com/science/article/pii/S0731708510000257
DOI
10.1016/j.jpba.2010.01.022
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Yoo, Tae Hyun(유태현) ORCID logo https://orcid.org/0000-0002-9183-4507
Ha, Sung Kyu(하성규)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/102060
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