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以从CHO工程细胞培液上清初步纯化的尿激酶型纤溶酶原激活剂(uPA)免疫BALB/c小鼠,通过杂交瘤技术制备出11株抗uPA单克隆抗体,经特异性实验表明,这些抗体特异性地识别高分子量uPA(5.4×10 ̄4u),与低分子量uPA(3.3×10 ̄4u)有不同程度结合,但都不识别1.8×10 ̄4u的uPA片段。它们与牛血清白蛋白、CHO细胞上清液及组织型纤溶酶原激活剂都无交叉反应。11株抗体分别针对5个不同的抗原决定簇。以单克隆抗体偶联的scpharose4B亲和柱直接从培液上清纯化uPA,并经sephacrv1S-200分子筛进一步纯化,纯度达95%以上,回收率为85%,纯化倍数为157倍左右。
BALB / c mice were immunized with urokinase-type plasminogen activator (uPA), which was purified from the supernatant of CHO engineering cell culture supernatant, and 11 monoclonal anti-uPA antibodies were prepared by hybridoma technique. . These antibodies specifically recognize high molecular weight uPA (5.4 × 10 ~ 4u) with different degrees of binding to low molecular weight uPA (3.3 × 10 ~ 4u) but do not recognize 1.8 × 10 ~ 4u uPA fragment. They did not cross-react with bovine serum albumin, CHO cell supernatants and tissue-type plasminogen activator. Eleven antibodies directed against five different epitopes. The uPA was directly purified from the supernatant of the culture by the monoclonal antibody conjugated scpharose4B affinity column and further purified by sephacrv1S-200 molecular sieve. The purity was over 95%. The recovery was 85% and the purification fold was about 157 times.