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1、中国农业科学2010,43(3):565-570ScientiaAgriculturaSinicadoi:10.3864/j.issn.0578-1752.2010.03.017半抗原-载体蛋白偶联物的三种电泳鉴定方法11，2311,秦毅兰11雷红涛，庞杰，贺丽苹，杨金易，孙远明，梁明标12（华南农业大学食品学院/广东省食品质量安全重点实验室，广州510642；福建农林大学食品科技学院，福州350002；3华南农业大学分析测试中心，广州510642）摘要：【目的】建立一种简单方便的半抗原-载体蛋白偶联物电泳鉴定方法。【方法】选用SDS-PAGE、非变性琼脂糖凝胶电泳、高效
2、毛细管区带电泳3种方法对载体蛋白、偶联剂变性的载体蛋白、半抗原-载体蛋白进行比较鉴定，同时与光谱法结果比较。【结果】对分子量差异较小的载体蛋白和半抗原-载体蛋白偶联物，SDS-PAGE不能有效分辨；但偶联前后载体电荷有变化时，非变性琼脂糖凝胶电泳却能有效分辨游离的载体蛋白和半抗原-载体蛋白偶联物，相对SDS-PAGE表现出更高的分辨灵敏度；毛细管区带电泳能高效分辨载体和半抗原-载体蛋白偶联物，结果直观，但需较贵重的专用仪器。琼脂糖电泳和毛细管电泳法与可见-紫外光谱法鉴定结果一致。【结论】3种电泳方法各有优缺点，非变性凝胶电泳简便、廉价、有效，较适合普通实验室使用。关键词：
3、半抗原-载体蛋白偶联物；SDS-PAGE；非变性琼脂糖凝胶电泳；毛细管区带电泳IdentificationofHapten-CarrierProteinConjugatebyThreeElectrophoresisMethods11,231111LEIHong-tao,PANGJie,HELi-ping,YANGJin-yi,SUNYuan-ming,QINYi-lan,LIANGMing-biao1（GuangdongKeyLaboratoryofFoodQualityandSafety/CollegeofFoodScience,SouthChinaAgricultur
4、alUniversity,2Guangzhou510642;CollegeofFoodScienceandTechnology,FujianAgricultureandForestryUniversity,Fuzhou350002;3InstrumentalAnalysisandResearchCenterofSouthChinaAgriculturalUniversity,Guangzhou510642）Abstract:【Objective】Toestablishasimpleelectrophoresischaracterizationmethodforhapten
5、-carrierproteinbioconjugate.【Method】SDS-PAGE,nondenaturingagarosegelelectrophoresisandhighperformancecapillaryelectrophoresiswereemployedandcomparedwithspectrummethod.【Result】TheresultshowedthatSDS-PAGEwasnotasensitivecandidatemethodtodifferentiatecarrierproteinfrombioconjugatewhenbothpro
6、teinshavelessmolecularweightdifference,butnondenaturinggelelectrophoresiswasabetteralternative,whichcoulddiscriminatethefreecarrierproteinandbioconjugateproteinwithdifferentnetchargenumbers.Highperformancecapillaryzoneelectrophoresiswasanotherpowerfulapproachforhapten-carrierproteinbiocon
7、jugatecharacterization.Theinformationobtainedfromthreeelectrophoresiswasinaccordancewiththeonefromultraviolet-visiblespectrumscanning.【Conclusion】Threeelectrophoresismethodshaddifferentadvantagesanddisadvantages,cost-savingandsimplenondenaturingelectrophoresisseemed
