摘要： 为对猪瘟病毒(CSFV)建立更加有效的临床检测方法.用纯化的猪瘟兔化弱毒免疫BALB/c小鼠,取其脾细胞与SP2/O骨髓瘤细胞融合.经ELISA方法筛选和3次亚克隆,最终获得了株能稳定传代并分泌抗猪瘟病毒单克隆抗体的杂交瘤细胞株,分别命名为:C7,C9,G9,G10和4E8,其分泌的单克隆抗体(McAb)为IgG1(G9,4E8)和IgG2a(C7.C9,G10)亚类.经鉴定,株单抗细胞培养上清效价为1:1 600～1:3200,腹水效价为1:1200～1:102400.交叉试验及特异性抗原阻断试验表明,所制备的McAb与其他抗原无交叉反应性,均完全针对猪瘟病毒(CSFV)抗原决定簇,具有高度特异性.稳定性试验表明,制备的杂交瘤细胞株经连续传代2代和经3次冻存复苏后,仍能稳定分泌特异性抗体.抗CSFV McAb的成功制备,为进一步研究CSFV的生物学特性及其快速诊断方法的建立奠定了基础.
关键词:
猪瘟病毒,
杂交瘤,
单克隆抗体,
ELISA,
鉴定
Abstract: The classical swine fever virus(CSFV)isolate was purified and used to vaccinate the BALB/ c mice. After immunized for 5 times,the mouse spleen cells were used to fuse with the myeloma cell line SP2/ 0.After screened by ELISA and subcloned for 3 times,five strains of hybridomas C7,C9,G9,G10 and 4E8 were obtained,of which G9 and 4E8 be2 longed to IgG1,while C7,C9 and G10 belonged to IgG2a.Among the five McAbs,the titers of cell cultures ranged from 1∶ 1 600 to 1∶ 3 200 and the titersof ascitesfluids ranged from 1∶ 51 200 to 1∶ 102 400. The preparation of these McAbs provid2 ed a basis for further studying the biological characteristics of CSFV and establishing rapid diagnostic method.
Key words:
CSFV,
Hybridoma,
Monoclonal antibody,
ELISA,
Identification
中图分类号:
许保疆, 陈陆, 游一, 刘春生, 郑鹿平, 王川庆. 抗猪瘟病毒单克隆抗体的制备及其生物学特性鉴定[J]. 华北农学报, 2009, 24(3): 64-68. doi: 10.7668/hbnxb.2009.03.014.
XU Bao jiang,, CHEN Lu, YoU Yi,, LIU Chun sheng, ZHENG Lu ping, WANG Chuan qing. Preparation of Monoclonal Antibodies against Classical Swine Fever Virus and Their Identification of Biological Characters[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(3): 64-68. doi: 10.7668/hbnxb.2009.03.014.