It is necessary to establish a rapid and accurate diagnostic method for controlling the epidemic of severe acute respiratory syndrome (SARS). Cooperating with Henan Bioengineering Research Center, we developed a new diagnostic kit for antibodies to SARS virus based on antigen-capturing ELISA. Here we report the clinical detection results obtained from several hospitals which used this diagnostic kit for SARS diagnosing. Comparing with other detection kits now used, the antigen-capturing ELISA has stronger specificity and higher sensitivity, and more fits for rapid confirming and diagnosing of SARS cases, SARS epidemiological investigation and SARS epidemic source research.
目的:制备抗人内皮抑素的单克隆抗体(mAb),为深入研究内皮抑素的作用机制提供一种有用的试剂。方法:以毕赤酵母表达的内皮抑索为免疫原免疫BALB/c小鼠,采用B细胞杂交瘤技术,以间接ELISA法筛选分泌抗人内皮抑素的特异性mAb的杂交瘤细胞株,并诱生腹水。腹水中的mAb采用 Protein A Sepharose CL-4B进行纯化。以标准的抗Ig血清做ELISA,鉴定mAb的Ig类、亚类及型;用免疫印迹法鉴定mAb的特异性。结果:获得1株可分泌抗人内皮抑素mAb的杂交瘤细胞株4E7,该株杂交细胞分泌的mAb属于IgGl(λ型)。腹水mAb的效价为1×10-4-1×10-6,纯化后大于1×10-10。免疫印迹结果显示,mAb 4E7可特异性地与酵母及大肠杆菌表达的内皮抑素起反应,而与bFGF等其他细胞因子不发生交叉反应。结论:分泌抗人内皮抑素mAb杂交瘤细胞株的建立,为进一步研究奠定了基础。
