为了制备TNF-α特异性抗体，用PCR的方法克隆出TNF-α片段的编码基因并将之插入PET-28a（+）原核表达载体，成功地在大肠杆菌中获得50%～60%的高效表达。经SDS-PAGE电泳，切下TNF-α特异条带，直接免疫大白兔，制备出TNF-α特异性抗体，抗体的效价可达1∶6400。利用该抗体成功地检测出mTNF-NIH3T3细胞膜上的跨膜型TNF-α的表达情况。结果表明：制备出的抗体具有较高的特异性，对TNF-α的免疫检测具有重要应用价值。%In order to prepare the specific antibody against soluble tumor necrosis factor α (sTNF-α), a gene fragment coding sTNF-α was cloned by using PCR and inserted into PET-28a(+) expression vector. Recombinant protein was expressed effectively in E.coli and separated by SDS-PAGE. By using the TNF-α specific band in SDS-PAGE agrose as antigen, the rabbits were immunized and the antibody against TNF-α was obtained with the titer being 1∶6400. The antibody was used to successfully detect the expression of transmembrane TNF-α on the cellular membrane of mTNF-NIH3T3. It was suggested that the antibody was high specific and was useful for immunological detection of TNF-α.
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