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目的制备和鉴定抗马尔尼菲青霉单克隆抗体(mAb)。方法用基因重组马尔尼菲青霉甘露糖蛋白1(MP1)抗原免疫BALB/c小鼠,制备mAb,并采用ELISA间接法和免疫印迹进行筛选和鉴定。结果筛选出4株稳定分泌抗马尔尼菲青霉mAb杂交瘤细胞株,IgG亚类鉴定均为IgG1,抗体亲和常数分别为8.2×10-9、4.7×10-9、6.5×10-9和2.7×10-9,免疫印迹证实马尔尼菲青霉mAb特异性识别MP1。结论4个杂交瘤细胞株特异性好、亲和力高,为马尔尼菲青霉病的快速诊断奠定了基础。
Objective To prepare and identify monoclonal antibodies against Penicillium marneffei (mAb). Methods BALB / c mice were immunized with the genetically modified Penicillium marneffei Mannoprotein 1 (MP1) antigen, and the mAbs were prepared and characterized by indirect ELISA and immunoblotting. Results Four strains of McAbs secreting mAbs against Penicillium marneffei were screened. The IgG subclasses were identified as IgG1. The antibody affinity constants were 8.2 × 10-9, 4.7 × 10-9 and 6.5 × 10-9, respectively And 2.7 × 10-9, immunoblotting confirmed that Penicillium marneffei mAb specifically recognized MP1. Conclusion The four hybridoma cell lines have good specificity and high affinity, which laid the foundation for the rapid diagnosis of Penicillium marneffei.