摘要
目的制备猪细小病毒(PPV)杂交瘤细胞株,并对其分泌的PPV单克隆抗体进行鉴定。方法按常规方法制备并获得2株杂交瘤细胞。用染色体分析对杂交瘤细胞进行鉴定,用间接ELISA、免疫过氧化物酶单层试验(IPMA)和间接免疫荧光试验(IFA)对其分泌的单克隆抗体进行效价测定、亚型鉴定和特异性鉴定。结果得到2株分泌单克隆抗体的杂交瘤细胞株2H9、1F9,染色体数目介于90～110之间。细胞上清效价均达1∶1×104,腹水效价均达1∶1×107,其亚型分别为IgG1、IgM,均为kappa链。2H9、1F9单抗与猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪伪狂犬病毒(PRV)、猪圆环病毒I型(PCV-1)、猪圆环病毒Ⅱ型(PCV-2)、乙脑病毒(JEV)等均无交叉反应。IPMA和IFA检测结果显示2H9、1F9单抗均能与接种于PK-15细胞的PPV发生特异性反应。结论成功制备了2株抗PPV杂交瘤细胞株,证实其产生的单克隆抗体具有良好的特异性和敏感性。
Objective Preparation and characterizing of porcine parvovirus（PPV） hybridomas cells and their monoclonal antibody.Methods Two hybridomas cells were obtained by conventional methods..The hybridomas cells were analysed and identified by chromosome analysis,indirect ELISA,the immunoperoxidase monolayer assay（IPMA） and indirect immunofluorescence（IFA） methods.Results Two hybridomas cells were obtained and named 2H9 and 1F9 respectively.The chromosome numbers were between 90 to 110.Titration of the hybridomas culture supernatant were 1∶1 × 104,and titration of ascitic fluid were 1∶1 × 107,their subclasses were IgG1,IgM,belonging to kappa chain,and without cross-reaction with PRRSV,CSFV,PRV,PCV-1,PCV2 and JEV.Furthermore,the specific reaction of antibodies and PPV in PK-15 cells was confirmed IPMA and IFA.Conclusion Two anti-PPV hybridomas cells were acquired and the characterizations of two monoclonal antibodies were confirmed.
出处
《中国比较医学杂志》
CAS
2011年第9期54-57,F0004,共5页
Chinese Journal of Comparative Medicine
基金
"十一五"国家科技支撑计划动物源性生物材料病毒检测技术研究(2008BAI54B06)
