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用 99%纯度的单链尿激酶型纤溶酶原激活剂 (scuPA )免疫BALB/c小鼠制备了 4株能稳定分泌抗scuPA的IgG1类单克隆抗体 (McAb )的杂交瘤细胞系G7、E8、D1和A9。检测了这四株单抗的特异性 ,它们与组织型纤溶酶原激活剂 (t PA )无反应 ,与双链尿激酶型纤溶酶原激活剂 (uPA )的结合力很弱。其中G7细胞株单抗与单、双链结合的差异最大。取纯化G7单抗制备亲和层析柱。用此柱直接从培养上清纯化scuPA ,得到的scuPA纯度为 96 3% ,回收率为 85 4% ,纯化倍数 5 0倍左右 ,比活 1 2 9× 10 5。纯化方法简单、操作方便、避免了因步骤繁杂造成的scuPA的降解 ,同时降低了成本。
Four BALB / c mice were immunized with 99% pure single-chain urokinase-type plasminogen activator (scuPA) to prepare four hybridoma cell lines, G7, which can stably secrete anti-scuPA monoclonal IgG1 antibody (McAb) E8, D1 and A9. The specificity of these four mAbs was tested and did not respond to the tissue-type plasminogen activator (t PA), but showed a weak binding to the urokinase-type plasminogen activator (uPA). G7 cell line monoclonal antibody and single, double-stranded combination of the largest difference. Affinity chromatography was performed using purified G7 mAb. The scuPA was directly purified from the culture supernatant by this column. The purity of scuPA was 96 3%, the recovery was 85 4%, the purification fold was about 50 times and the specific activity was 1 29 × 105. The purification method is simple, the operation is convenient, the scuPA degradation caused by the complicated steps is avoided, and the cost is reduced at the same time.