title: Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of 6-benzylaminopurine and its ribose adduct in bean sprouts.
abstract: 6-Benzylaminopurine (6-BA), a cytokinin plant growth regulator, has been banned for use in bean sprout production in China. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed with a specific monoclonal antibody (mAb 3E5). The assay showed a half-maximum inhibition concentration (IC50) and detection range of 18.9 ng/mL and 3.6-106 ng/mL, respectively. Recoveries of 6-BA spiked in home cultured bean sprout samples averaged from 75% to 89% with a correlation coefficient (R(2)) of 0.998 between the results determined by icELISA and those by liquid chromatography-electrospray ionization quadrupole Orbitrap mass spectrometry (LC-ESI-MS). LC-ESI-MS showed that 6-BA had been partially metabolized to 6-benzylaminopurine riboside (6-BAR) in the positive samples. The content of 6-BA determined by icELISA was about 5-70 times higher than that of LC-ESI-MS because mAb 3E5 had 315% cross-reactivity with 6-BAR. Such icELISA being ultra-sensitive to 6-BAR would allow quick monitoring of 6-BA by detecting 6-BAR as a potential biomarker.
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Lishan
authors.initials: L
authors.lastname: He
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Suqin
authors.initials: S
authors.lastname: Guo
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Huanfang
authors.initials: H
authors.lastname: Yue
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
authors.forename: Xiangxue
authors.initials: X
authors.lastname: Ning
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China. Electronic address: tanguiyu@cau.edu.cn.
authors.forename: Guiyu
authors.initials: G
authors.lastname: Tan
authors.affiliation: Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI 96822, USA.
authors.forename: Qing X
authors.initials: QX
authors.lastname: Li
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China. Electronic address: wbaomin@263.net.
authors.forename: Baomin
authors.initials: B
authors.lastname: Wang
keywords: 6-Benzylaminopurine; 6-Benzylaminopurine (PubChem CID: 62389); 6-Benzylaminopurine riboside (PubChem CID: 92208); Benzyladenosine (PubChem CID: 250699); ELISA; Food safety; Monoclonal antibody; Plant growth regulator