title: Bispecific Monoclonal Antibody-Based Multianalyte ELISA for Furaltadone Metabolite, Malachite Green, and Leucomalachite Green in Aquatic Products.
abstract: A new multianalyte immunoassay was designed to screen furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ), malachite green (MG), and leucomalachite green (LMG) in aquatic products using a bispecific monoclonal antibody (BsMAb). Gradient drug mutagenesis methods were separately used to prepare an anti-3-nitrobenzaldehyde-derivatized AMOZ (3-NPAMOZ) hybridoma cell line that was hypoxanthine-guanine-phosphoribosyltransferase (HGRPT) deficient and an anti-LMG hybridoma cell line that was thymidine kinase (TK) deficient. BsMAb recognizing 3-NPAMOZ and LMG was generated using hybrid-hybridomas of HGRPT and TK deficient cell lines. For AMOZ and LMG, respectively, the BsMAb-based indirect competitive ELSIA (ic-ELISA) values of 1.7 ng/mL and 45.3 ng/mL and detection limits of 0.2 ng/mL and 4.8 ng/mL. To establish the ic-ELISA, 3-NPAMOZ derivatized from AMOZ with 3-nitrobenzaldehyde and LMG reduced from MG by potassium borohydride was recognized by BsMAb. Recoveries of AMOZ, MG, and LMG in aquatic products were satisfactory and correlated with HPLC analysis. Thus, the multianalyte ic-ELISA is suitable for rapid quantification of AMOZ, MG, and LMG in aquatic products.
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Yu-Dong
authors.initials: YD
authors.lastname: Shen
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.|Zhongshan Quality Supervision & Inspection Institute of Agricultural Products , Zhongshan 528403, China.
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Li
authors.affiliation: Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California , Davis, California 95616, United States.
authors.forename: Jie-Xian
authors.initials: JX
authors.lastname: Dong
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Zhen-Lin
authors.initials: ZL
authors.lastname: Xu
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Yang
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Yuan-Ming
authors.initials: YM
authors.lastname: Sun
authors.affiliation: Guangdong Provincial Key Laboratory of Food Quality and Safety, College of Food Science, South China Agricultural University , Guangzhou 510642, China.
authors.forename: Zhi-Li
authors.initials: ZL
authors.lastname: Xiao
keywords: ELISA; bispecific monoclonal antibody; furaltadone metabolite; leucomalachite green; malachite green; multianalyte immunoassay