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Development of Immunochip for Detection of Clenbuterol Residue |
SUN Lai-yu,QIAN Kun,SHAO Chao-gang,YIN Li,YAN Qiong-qiong, LU Yun-hua,YANG Jin-tian |
School of Life Science, Huzhou University, Huzhou, Zhejiang 313000, China |
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Abstract Immunochip is designed in order to realize the massive detection of clenbuterol residue.Slide glass surface as the carrier is modified by APTES-DGA.The clenbuterol antibody is coupled to the slide glass surface.Glass surface active groups are blocked by bovine serum albumin. The samples and standards, horseradish peroxidase labeled clenbuterol are added into the immunochip.CLB and CLB-HRP are combined competitively with Ab (CLB).Chemiluminescence reaction is catalyzed by the horseradish peroxidase and optical signal value is detected Sampling volume, incubation temperature, incubation time, concentration of washing solution, chemiluminescence integral time are tested.The results show that glass slide with APTESDGA has a better signal uniformity and antibody coupling ability. In the detection process, immunochip signal is stable when sampling volume is 50 μL, incubation temperature is 37 ℃, incubation time is 30 min, wash solution concentration is 10 fold diluent and chemiluminescence reader integral time is 60 s. The method is fast, simple and accurate by immunochip analysis for clenbuterol residue.
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[1]孙来玉,尹莉,姚婷,等 盐酸克伦特罗残留检测研究进展[J]. 湖州师范学院学报, 2014,36(4):27-32.
[2]Zhuang Y, Yue C, Xie F.Fluorimetric method based on diazotizationcoupling reaction for determination of clenbuterol [J].J Fluoresc, 2014,24(3):945-950.
[3]Engelmann M D, Hinz D, Wenclawiak B W- Solid-phase micro extraction (SPME) and headspace derivatization of clenbuterol followed by GC-FID and GCSIMMS quantification[J].Anal Bioanal Chem, 2003, 375(3) : 460-464.
[4]Ana G A, Pablo R G, Iván L, et al. Development of a routine method for the simultaneous confirmation and determination of clenbuterol in urine by minimal labeling isotope pattern deconvolution and GC-EI-MS[J]. Anal Bioanal Chem,2012, 402(5):1879-1888.
[5]Li Y, Hu J T, Shi Y, et al.Simultaneous determination of seven adulterants in slimming functional foods by HPLCESIMS/MS[J]. Food Anal Methods, 2011, 4(4):505-516.
[6]Zhen G H, Deng L G, Lu X, et al. UPLC-ESI-MS-MS determination of three β2-agonists in pork[J]Chromatographia, 2010, 72 (1-2): 79-84.
[7]Vela J, Yanes E G, Stalcup A M Quantitative determination of clenbuterol, salbutamol and tulobuterol enantiomers by capillary electrophoresis[J]. Fresenius J Anal Chem,2001,369(3-4) :212-219.
[8]Jiao F, Chen X, Hu Y, et al. Enantiomeric separation of racemic clenbuterol by capillary zone electrophoresis[J]. J Iran Chem Soc, 2008, 5(4):553-558.
[9]Zhang M Z, Wang M Z, Chen Z L, et al. Development of a colloidal goldbased lateralflow immunoassay for the rapid simultaneous detection of clenbuterol and ractopamine in swine urine[J]. Anal Bioanal Chem, 2009,395(8):2591-2599.
[10Liu X X, Wang H, Liang Y,et al. Production and characterization of a single-chain Fv antibody-alkaline phosphatase fusion protein specific for clenbuterol[J].Mol Biotechnol,2010,45(1):56-64.
[11]Reena R Gaichore, Srivastava A K, et al.Multiwalled carbon nanotube-4-tert-butyl calixarene composite electrochemical sensor for clenbuterol hydrochloride determination by means of differential pulse adsorptive stripping voltammetry[J].J Appl Electrochem, 2012, 42(9):979-987.
[12]Julien B T, Masson J F. Nanostructured substrates for portable and miniature SPR biosensors[J]. Anal Bioanal Chem, 2012, 403(6):1477-1484.
[13]Yang Y P, Lei X Y, Yue A, et al. Temperaturedependent THz vibrational spectra of clenbuterol hydrochloride[J].Sci China-Phys Mech Astron, 2013, 56( 4):713-717.
[14]Maureen D M, Lara B C, Olga G,et al.Immunochip analysis identifies multiple susceptibility loci for systemic sclerosis[J]. Am J Hum Genet, 2014, 94(1): 47-61.
[15]孙来玉,尹莉,姚婷,等. 瘦肉精克伦特罗残留检测免疫芯片技术的研究[J]. 药物分析,2014,34(5):859-864.
[16]刘天龙,王燕飞,邹明强,等.牛奶中抗生素的液相芯片检测方法研究[J]. 计量学报, 2010,31(5A):29-32.
[17]Michael V, Rudolf O, Ellen U. A platelet protein biochip rapidly detects an Alzheimer’s diseasespecific phenotype[J]. Acta Neuropathol, 2014; 128(5): 665-677
[18]Gehring A, Barnett C, Ted C, et al. A highthroughput antibody-based microarray typing platform[J].Sensors,2013 ,13(5): 5737-5748.
[19]吴坚,任艺.生物微流控PCR荧光芯片微通道动态检测系统研究[J]. 计量学报, 2007,28(3A):284-286.
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