A sensitive and reliable quantification method for Bisphenol A based on modified competitive ELISA method.
Bisphenol A (BPA), generally known as bisphenols, has been identified as a potential estrogenic substance. BPA must be conjugated to carrier protein and BSA was commonly used. 4,4-Bis(4-hydroxyphenyl) valeric acid (BHPVA) has a bisphenolic structure and a long carbon chain with a reactive carboxyl group on the end. In this study, BHPVA-BSA was used to produce polyclonal antibody against bisphenolic structure, and a modified competitive ELISA method for quantification of BPA was developed. This system was based on BHPVA-BSA for polyclonal antibody production against bisphenolic structure, and BHPVA-HRP for determination of BPA substituting detection antibody in competitive reaction. Recovery was assessed at 10 different concentrations (2-1000 ng/ml) of BHPVA, and the recovery range was from 96.3% to 107.2%. The variation was from 6.2% to 9.8% for intra assay and from 10.1% to 12.6% for inter assay. The quadratic was used to establish the curve regression. The range was found to be between 2 and 1000 ng/ml. This modified competitive ELISA method has proven to be a very useful tool for quantification of BPA without the unexpected interaction of BSA and anti-BSA polyclonal antibody.
Kim
Andre
A
Department of Chemistry and Centre for Innovative Bio-physio sensor Technology, Pusan National University, Busan 609-735, Republic of Korea.
Li
Chun-Ri
CR
Jin
Chun-Feng
CF
Lee
Keun Woo
KW
Lee
Sang-Hoon
SH
Shon
Kwang-Jae
KJ
Park
Nam Gyu
NG
Kim
Dong-Kyoo
DK
Kang
Shin-Won
SW
Shim
Yoon-Bo
YB
Park
Jang-Su
JS
D001559:Benzhydryl Compounds; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015394:Molecular Structure; D010636:Phenols / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D000098826:Bisphenol A Compounds
D001559:Benzhydryl Compounds; D010636:Phenols; C006780:bisphenol A; D000098826:Bisphenol A Compounds