摘要
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 (mAbs) against Penicillium marneffei. Methods Recombinant mannoprotein1 (MP1) of Penicillium marneffei was used to immunize BALB/c mice, and anti-MP1 mAbs were obtained by means of hybridoma. Screening and identification of the mAbs were subsequently performed with indirect enzyme-linked immunosorbent assay and Western blotting, respectively. Results Four hybridomas producing antibodies against Penicillium marneffei were obtained, and the IgG isotypes of the 4 mAbs were identified as IgG1 with affinity constants (K) of 8.2×10 -9 , 4.7×10 -9 , 6.5×10 -9 and 2.7×10 -9 , respectively. Western blotting demonstrated specific recognition of Aspergillus fumigatus MP1 by the obtained mAbs. Conclusions The 4 hybridomas producing anti-MP1 mAbs with high specificity and affinity can be of significant value in the diagnosis of Penicilliosis marneffei infections.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第1期38-39,52,共3页
Journal of First Military Medical University
基金
第一军医大学珠江医院与香港大学合作项目~
关键词
马尔尼菲青霉素
单克隆抗体
制备
鉴定
Penicillium marneffei
mannoprotein
monoclonal antibody
enzyme-linked immunosorbent assay
