卡那霉素人工抗原的制备及鉴定-中国农学通报.pdf

中国农学通报 2014,30(17):26-30 Chinese Agricultural Science Bulletin 卡那霉素人工抗原的制备及鉴定 王爱萍，栗 宁，祁艳华，段倩倩，郑 伟，王新洲，裴艳艳，周景明 （郑州大学生命科学学院，郑州450001 ） 摘 要：为了人工制备卡那霉素完全抗原。将卡那霉素标准品与牛血清白蛋白(BSA)或卵清白蛋白 (OVA)偶联，制备人工抗原。利用两种不同的偶联方法：碳二亚胺法(EDC)和戊二醛法(GDA) ，分别制备 出KAN-BSA （作为免疫原），KAN-GDA-OVA （作为包被原）。利用SDS凝胶电泳鉴定和动物免 疫试验等，鉴定了完全抗原偶联情况。结果显示：制备的卡那霉素完全抗原能刺激小鼠机体产生相应抗 4 体，且抗血清效价均达到1:2.56×10 ，其中一号小鼠的IC50 值为22.28 ng/mL ，说明完全抗原免疫原性良 好，抗原制备成功。 关键词：卡那霉素；人工抗原；碳二亚胺法；戊二醛法 中图分类号：S8 文献标志码：A 论文编号：2014-3318 SynthesisandIdentificationofKanamycinCompleteAntigenandPreparationof PolyclonalAntibodyAgainstKanamycin WangAiping,LiNing,QiYanhua,DuanQianqian,ZhengWei,WangXinzhou,PeiYanyan,ZhouJingming (ZhengzhouUniversity,Zhengzhou450001) Abstract: Thepurposeofthisexperimentwaspreparingkanamycincompletelyartificialantigens.Firstly, kanamycinstandardwasreactedwithbovineserumalbumin(BSA)orovalbumin(OVA)topreparekanamycin artificial antigens. Two different coupling methods carbodiimide (EDC) and glutaraldehyde (GDA) were respectivelypreparedKAN-BSA(asanimmunogen)andKAN-GDA-OVA(asthecoatingantigen).Bothof theSDSgelelectrophoresisandtheanimalimmunizationexperimentidentifiedtheantigen-coupled case.Theresultsshowedthat,thepreparedkanamycincompleteantigencouldstimulatethebodiestoproduce antibodiesinmice,andtheantiserumtiterreached 1:2.56 × 10,TheIC ofthemicewas 22.28 ng/mL.The4 50 resultsillustratedtheimmunogenicityofthecompleteantigensweregoodandthepreparationsoftheantigens weresuccessful. Keywords:Kanamycin;artificialantigen;carbodiimidemethod;glutara