title: Monoclonal antibody production and indirect competitive enzyme-linked immunosorbent assay development of 3-methyl-quinoxaline-2-carboxylic acid based on novel haptens.
abstract: Two novel immunizing haptens of 3-methyl-quinoxaline-2-carboxylic acid (MQCA) were synthesized and conjugated with cationized bovine serum albumin. Female BALB/c mice were immunized with above conjugates, splenocytes were fused with Sp2/0 cells to produce monoclonal antibody. Compared with previous studies, antibodies raised in this work showed higher sensitivity. Meantime, a novel heterologous coating hapten was also prepared. The indirect competitive enzyme-linked immunosorbent assay (icELISA) based on the optimum condition showed an IC50 of 3.1μg/kg (ppb), and the linear range of 0.46-10.5ppb for MQCA. The limit of detect (LOD) of MQCA in swine muscle, swine liver and chicken was 0.32, 0.54, and 0.28ppb, respectively. The LOD of this assay can satisfy the minimum required performance levels (4ppb) for MQCA. These results indicated that the proposed ELISA, with high sensitivity and specificity, as well as good reproducibility and accuracy, is suitable for determination of MQCA residues in food samples.
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China.
authors.forename: Guopeng
authors.initials: G
authors.lastname: Li
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China. Electronic address: zf@cau.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhou
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China.
authors.forename: Jiaying
authors.initials: J
authors.lastname: Li
authors.affiliation: Beijing Weifengyongzhu Science and Technology Development Co., LTD, District A, Yuanmo Technology Industrial Park, Changping, Beijing 100020, People's Republic of China; Pingyuan Weifengyongzhu Science and Technology Co., LTD, Depinglianjiexian West Street, Pingyuan District, Dezhou City, Shandong Province 253100, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Xing
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China.
authors.forename: Tiangang
authors.initials: T
authors.lastname: Wang
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China.
authors.forename: Xilong
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China.
authors.forename: Baoping
authors.initials: B
authors.lastname: Ji
authors.affiliation: Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Tsinghua East Road 17, Haidian District, Beijing 100083, People's Republic of China; China Development Bank, Fuxingmennei Street 18, Xicheng District, Beijing 100031, People's Republic of China.
authors.forename: Wanpeng
authors.initials: W
authors.lastname: Ren
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000917:Antibody Formation; D002645:Chickens / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005506:Food Contamination / Q000032:analysis; D006241:Haptens / Q000737:chemistry* / Q000276:immunology; D007114:Immunization; D018796:Immunoconjugates; D057230:Limit of Detection; D008460:Meat / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D011810:Quinoxalines / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine / Q000378:metabolism
chemical_list: C581287:3-methylquinoxaline-2-carboxylic acid; D000911:Antibodies, Monoclonal; D006241:Haptens; D018796:Immunoconjugates; D011810:Quinoxalines
keywords: 3-Methyl-quinoxaline-2-carboxylic acid; ELISA; Hapten; Monoclonal antibody