犬细小病毒荧光抗体的制备及其鉴定.pdfVIP

犬细小病毒荧光抗体的制备及其鉴定 高丽啦杜莹憎 张忠湛坩 杨松涛2 夏咸柱2‘ 高玉伟2 (1．吉林大学畜牧兽医学院长春130062； 2．解放军军事医学科学院军事兽医研究所长春130062) 摘要：采用常规方法建立杂交瘤细胞株并制备抗犬细小病毒(Canineparvovirus) VP2蛋白单克隆抗体(MAb)；小鼠腹腔注射阳性克隆生产腹水；以C蛋白亲和层析法 G一25去除游离荧 进行纯化；用透析法对单抗进行FITC(异硫氰酸荧光素)标}己；Sephadex 光索；测定F／P值及荧光抗体工作浓度；以吸收、阻断、抗原交叉试验对荧光抗体进行特异 性鉴定。结果表明，获得2株能稳定分泌CPV单克隆抗体的杂交瘤细胞株，选取3G4单抗 成功地制备了质量较高的荧光抗体。 关键词：犬细小病毒；单克隆抗体；荧光抗体 and of Preparationpreliminaryapplication fluorescent Canine antibodyagainst parvovirus GAOLil·2DU XIAXian—zhu2+GAOYu—wei2 Yin91”ZHANGZhong—zhan。”YANGSong—ta02 of of MedicalSciencesof (1．Institute 130062，China； MilitaryVeterinary，AcademyMilitary PLA，Changchun ofAnimalScienceand Medicine．Jilin 1313062，China) 2．College Veterinary University，ChangChun obtainfluoescent canine a- Abstract：To antibodyagainst parvovirus，monoclonal antibody was means VP2 ofCPV oftheconventional was gainstprotein developedby protoc01．Ascitespro-- duced coloniesand G anti— usingpositive purifiedbyproteinaffinity waslabelledwithfluorescein body isothiocyanate(FITC)usingdialysis and F-／P FITCwasremoved G-25columnthe valueand dilutionof bySephadex optimalworking fluorescent testand inter