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RHDVVLPs单克隆抗体的制备及其识别表位的初步定位

DOI: 10.7668/hbnxb.2015.05.011, PP. 65-70

Keywords: RHDVVLPs,单克隆抗体,表位,初步定位

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Abstract:

为了筛选兔出血症病毒(RHDV)衣壳蛋白(VP60)的特异性单抗,并进一步分析单抗识别表位的分布情况,将重组杆状病毒rAcV-Bac-VP60接种Sf9昆虫细胞,收获细胞培养物,电镜观察显示重组VP60蛋白获得有效表达,并可组装成病毒样粒子(VLPs)。将RHDVVLPs作为免疫原与等量弗氏佐剂乳化,免疫BALB/c小鼠,取脾细胞与SP2/0骨髓瘤细胞融合,经3次亚克隆后,获得11株能稳定分泌抗RHDVVLPs抗体的阳性杂交瘤细胞株。间接ELISA、IFA和WesternBlot鉴定结果表明,这11株单克隆抗体均能够特异地识别RHDVVLPs和天然RHDV。11株单抗均为IgG1,其中1D4和3F7的轻链为Lambda型,其余均为Kappa型。截短表达结果显示,单抗5A3针对RHDVVP60的NTA区,5F3针对RHDVVP60的S区,单抗1B8、1D4、3D11、3F7、4C2、4G2、5G2、5H3和6B2针对RHDVVP60的P区。研究为RHDVVLPs抗原表位的鉴定和RHDV结构功能的研究奠定了基础,同时为RHDV的检测和新型疫苗研究提供物质基础。

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