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莱克多巴胺人工抗原及抗体的制备 被引量:5

Preparation of Ractopamine Artificial Antigen and Antibody
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摘要 莱克多巴胺是一种禁用β-兴奋剂,建立莱克多巴胺药残的快速检测方法是实现对其进行有力监控的有效途径,而莱克多巴胺抗体是快速免疫检测法的基本试剂。本实验用全新的方法研究了莱克多巴胺免疫原的合成,采用对氨基苯甲酸(ABA)和1,4-丁二醇缩水甘油醚(BDE)将莱克多巴胺分别和cBSA、cOVA偶联,合成了莱克多巴胺的免疫原和包被抗原,并对其进行了紫外分析。用免疫原免疫新西兰大白兔,获得了高灵敏度和特异性的莱克多巴胺多克隆抗体,采用间接ELISA法检测抗体的IC50值为4.34ng/ml,所得多克隆抗体效价达到102400。 As one of β-agonists, ractopamine has been prohibited in food production and animal husbandry. To enforce regulation, it is necessary to develop a rapid detection system to measure the residue of ractopamine in animal food production. In this study, a new ractopamine immunogen was synthesized by using 4-aminobenzoic acid as a linker to connect hapten with carrier protein, bovine serum albumin (BSA). Besides the immunogen, a new ractopamine coating antigen was synthesized also by using 1,4-butanediol diglycidyl ether (BDE) as a linker to connect ovalbumin (OVA). The BSA conjugate was used to immunize New Zealand rabbits and polyclonal antibodies against ractopamine were obtained. The titers of antibodies were up to 102400 by indirect ELISA and the antibodies showed high sensitivity in the heterologous assay using RAC-BDE-cOVA as coating antigen, with an IC50 value of 4.34 ng/ml. The artificial antigen has perfect immunogencity and the obtained antibody has excellent sensitivity.
出处 《食品科学》 EI CAS CSCD 北大核心 2009年第9期25-28,共4页 Food Science
基金 青岛市科技计划项目(06-3-2-4-nsh) 国家自然科学基金项目(20675048) 国家“863”计划项目(07AA10Z435 2007AA06A407)
关键词 莱克多巴胺 ELISA 偶联 免疫原 ractopamine ELISA conjugation antigen
作者简介 孙海新(1978-),男,工程师,硕士,主要从事兽用生物制品研发及动物源性食品的安全研究。E-mail:hisense_sun@sina.com 通讯作者:郗日沫(1962-),男,教授,博士,主要从事生物免疫检测技术研究。E—mail:xirim02000@yahoo.com.cn
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  • 1GABIOLA C, CALONGE M A G, PORTILLO M P, et al. Validation of a method for the determination of salbutamol in animal urine by gas chromatography-mass spectrometry and its application to treated lamb samples[J]. J Microcolumn Separations, 1996, 8: 361-364.
  • 2SHELVER W L, SIMINTH D J, BERRY E S. Production and characterization of a monoclonal antibody against the β-adrenergic agonist ractopamine[J]. Journal of Agricultural and Food Chemistry, 2000, 48: 4020-4026.
  • 3SHELVER W L, SMITH D J. Determination of ractopamine in cattle and sheep urine samples using an optical biosensor analysis: comparative study with HPLC and ELISA[J]. Journal of Agricultural and Food Chemistry, 2003, 51: 3751-3721.
  • 4VANOOSTHUYZE K E I, ARTS C J M, PETEGHEM C H V. Development of a fast and simple method for determination of β -agonists in urine by extraction on empore membranes and detection by a test strip immunoassay[J]. Journal of Agricultural and Food Chemistry, 1997, 45: 3129-3137.
  • 5REGUEIRO J A G, PEREZ B, CASADMONT G. Determination of clenbuterol and salbutamol in urine by capillary gas chromatography with capillary columns of 1001am[J]. J Chromatogr A, 1993, 655: 73- 76.
  • 6SHEN S H, OYANG J, BAEYENS W R G, et al. Determination of β- agonists by ion chromatography with direct conductivity detection [J]. J Pharmaceutical and Biomedical Analysis, 2005, 38: 166-172.
  • 7POLEITINI A, MONTAGNA M, HONGENDOORN E A, et al. Applica-bility of coupled-column liquid chromatography to the analysis of β- agonists in urine by direct sample injection 1. Development of a singleresidue reversed-phase liquid chromatography-UV method for clenbuterol and selection of chromatographic conditions suitable for multi-residue analysis[J]. J Chromatogr A, 1995, 695: 19-31.
  • 8SPYRIDAKI M H, VONAPARTI P K A, VALAVIANI P. et al. Doping control analysis in human urine by liquid chromatography-electrospray ionization ion trap mass spectrometry for the Olympic Games Athens 2004: Determination of corticosteroids and quantification of ephedrines, salbutamol and morphine[J]. Anal Claim Acta, 2006, 573: 242-249.
  • 9JOSEFSSON M, SABANOVIC A. Sample preparation on polymeric solid phase extraction sorbents for liquid chromatographic-tandem mass spectrometric analysis of human whole blood-A study on a number of beta-agonists and beta-antagonists[J]. J Chromatogr A, 2006, 1120: 1- 12.
  • 10YAMAMOTO I, IWATA K. Enzyme immunoassay for clenbuterol, an β-adrenergic stimulant[J]. J Immtmoassay, 1982, 3: 155-171.

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