Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

被引:46
作者
He, Jinxin [1 ]
Wang, Yuan [1 ]
Zhang, Xiaoying [1 ]
机构
[1] Northwest A&F Univ, Coll Vet Med, Yangling 712100, Shaanxi, Peoples R China
关键词
Kanamycin; Egg yolk antibody (IgY); ELISA; Drug residues; FREE ELECTROCHEMICAL IMMUNOSENSOR; LINKED-IMMUNOSORBENT-ASSAY; AMINOGLYCOSIDE ANTIBIOTICS; IMMUNOGLOBULIN IGY; MILK; IMMUNOASSAY; GENTAMICIN; APTASENSOR; ANTIBODIES;
D O I
10.1007/s12161-015-0248-x
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
The present study was aimed to evaluate the feasibility to use chicken egg yolk antibodies (IgY) for the detection of Kanamycin (Kana) residues in food stuffs. Bovine serum albumin (BSA) was conjugated with Kana by carbodiimide method, and the laying chickens were immunized with Kana-BSA conjugate. PEG-6000 method was employed to extract IgY. The peak titer of anti-Kana IgY was 1:256,000 after the fifth booster immunization. The optimal dilution for anti-Kana IgY was 1:25,000 to obtain optical density (OD) of 1.0 at 2 mu g/mL of OVA-Kana coating concentration. The indirect competitive ELISA (ic-ELISA) showed that the IC50 value of anti-Kana IgY was 4.48 ng/mL and the regression curve equation was y = -15.62x + 60.17 (R (2) = 0.98, n = 3). The ic-ELISA showed a lower cross-reactivity (less than 0.01 %) with other drugs (except gentamicin-1.91 %). Recoveries from milk, pork, and chicken powder samples were in the range of 82.02 to 98.20 %, with relative standard deviation lower than 5.13 %. Our results indicate that generated anti-Kana IgY can be used in routine screening analysis of Kana residues in food samples.
引用
收藏
页码:744 / 751
页数:8
相关论文
共 27 条
[1]  
[Anonymous], 2003, EMEAMRL88603FINAL
[2]   Label-free detection of kanamycin using aptamer-based cantilever array sensor [J].
Bai, Xiaojing ;
Hou, Hui ;
Zhang, Bailin ;
Tang, Jilin .
BIOSENSORS & BIOELECTRONICS, 2014, 56 :112-116
[3]   Assay of Kanamycin A by HPLC with Direct UV Detection [J].
Blanchaert, B. ;
Jorge, E. Poderos ;
Jankovics, P. ;
Adams, E. ;
Van Schepdael, Ann .
CHROMATOGRAPHIA, 2013, 76 (21-22) :1505-1512
[4]   Detection of alkaline phosphatase by competitive indirect ELISA using immunoglobulin in yolk (IgY) specific against bovine milk alkaline phosphatase [J].
Chen, CC ;
Tai, YC ;
Shen, SC ;
Tu, YY ;
Wu, MC ;
Chang, HM .
FOOD CHEMISTRY, 2006, 95 (02) :213-220
[5]   Immunosensor based on magnetic relaxation switch and biotin-streptavidin system for the detection of Kanamycin in milk [J].
Chen, Yi Ping ;
Zou, Ming Qiang ;
Qi, Cai ;
Xie, Meng-Xia ;
Wang, Da-Ning ;
Wang, Yan-Fei ;
Xue, Qiang ;
Li, Jin-Feng ;
Chen, Yan .
BIOSENSORS & BIOELECTRONICS, 2013, 39 (01) :112-117
[6]   Rapid enzyme-linked immunosorbent assay and colloidal gold immunoassay for kanamycin and tobramacin in swine tissues [J].
Chen, Yiqiang ;
Wang, Zhiqin ;
Wang, Zhanhui ;
Tang, Shusheng ;
Zhu, Yan ;
Xiaos, Xilong .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2008, 56 (09) :2944-2952
[7]   Preparation of Artificial Antigen and Egg Yolk-derived Immunoglobulin (IgY) of Citrinin for Enzyme-Linked Immunosorbent Assay [J].
Duan, Zhao-Hui ;
Lin, Zhuang-Sen ;
Yao, He-Rui ;
Gao, Yan-Hong ;
Zhang, Kun ;
Zhao, Su-Qing ;
Zhu, Zhen-Yu .
BIOMEDICAL AND ENVIRONMENTAL SCIENCES, 2009, 22 (03) :237-243
[8]  
Isoherranen N, 1999, J AOAC INT, V82, P1017
[9]   Aptamers as therapeutics [J].
Keefe, Anthony D. ;
Pai, Supriya ;
Ellington, Andrew .
NATURE REVIEWS DRUG DISCOVERY, 2010, 9 (07) :537-550
[10]   Development and immunochemical evaluation of antibodies Y for the poorly immunogenic polypeptide prothymosin alpha [J].
Klimentzou, P ;
Paravatou-Petsotas, M ;
Zikos, C ;
Beck, A ;
Skopeliti, M ;
Czarnecki, J ;
Tsitsilonis, O ;
Voelter, W ;
Livaniou, E ;
Evangelatos, GP .
PEPTIDES, 2006, 27 (01) :183-193