抗恩諾沙星單克隆抗體的研制及初步應用.pdf

1、西南農(nóng)業(yè)大學碩士學位論文抗恩諾沙星單克隆抗體的研制及初步應用姓名：李海申請學位級別：碩士專業(yè)：臨床獸醫(yī)學指導教師：彭遠義20050501兩南農(nóng)業(yè)大學碩士論文ABSTRACT寰鼉置I————————————————————————————————————————————一——————皇I量量蔓■曼|毫曼曼曼曼曼PreparationandApplicationofMonoclonalAntibodyagainstEnrofloxacinC

2、andidate：LiHaiSupervisor：PengYuanyi(CollegeofAnimal＆Technology，SouthwestAgriculturalUniversity。Chongqing400716)AbstractEnrofloxacin(ENR)，afluoroquinolone，isspecificfortreatmentandpreventionofanimaldisenseRecently，however

3、，duetowide—ranginguseofENRresultinginresidueiSanimaltissue。ithasbeengivengreatinterestAndtheuseofENRinveterinarymedicinehasbeenprohibitedinmanycountriesInadditionithasapprovedbyEuropeaaCommunitythattheMaximumResiduelimit

4、s(MRL)ofENRinedibleanimaltissuearenotbeyond30ug／kgThepurposeofourstudyistoprepareofMonoclonalAntibody，toestablishthemethodofindirectcompetitiveELISA(CiELISA)detectingENRresidueandtomakefoundationforexplorationandapplicat

5、ionoftheENRresiduetestkitinthisstudy，ENRwascoupledtoBSAandOVAcarrierproteinrespectivelybyNhydroxysuccinimideactivatedestermethodbywhichENR‘‘BSAandENR——OVAwerepreparedBothantigenswereidentifiedbycOlourreactionofFeel3andUV

6、scanTheresultswereasfollows：exteriorsolutionindialysiSbecamelightyellowwhileinteriorsolutionappearedredresultfromreactionofFecl3ThemaximumabsorbancepeaksofENR—BSAandENR—OVAwereat319nmand320nmwavelength，respectivelyAndthe

7、rewereacommonDeakat332nmwavelengthforthetwoantigensThedetectionindicatedwhetherabsorbancespectrumofENRBSAorENR—OVAwasintegrationofspectrumofENRandpratein，whichincludedthecharacteristicabsorbancepeakofENRENR—BSAwasusedasi

8、mmunogentoimmunize8一weekoldBALB／CmiceforobtainingofantiseraENR(3VAwasusedascoatingantigenTheposjtivemiceofhightitreandantiseraagainstENRwereselectedbytheIndirectEnzyme1inkedI咖unosorbentAssay(ELSA)Splenoeytesfromthemicewe

9、refusedwithsp2／0myelomacellsThroughdetermining，cloningandselecting，B6andG12secretingmonoclonalantibodiesagainstENRwereselectedTheidentificationofsuhclassesindicatedthatthemonoclonalantibodiessecretedbyB6andG“wereantibodi