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提出一种新型制备电泳方法即多通道流动电动电泳技术,建立了微机控制的电泳设备,考查了操作条件对设备分离效率的影响。应用该技术进行牛血清清蛋白(BSA)-牛血红蛋白(HBB)混合物的连续分离,每小时从含BSA、HBB各37.0mg的蛋白质混合物中分离出13.6mg的BSA和20.0mg的HBB。应用该技术进行尿激酶粗品的纯化,可将其比活力由4000IU/mg提高到4×10~4IU/mg以上,产量在10×10~4IU/h以上。将该技术与(NH_4)SO_4沉淀方法结合,从15ml免疫鼠血清中分离出12.5mg高纯度尿激酶抗体IgG,产量为2.1mg/h。上述结果证明多通道流动电泳技术具有分离速度快、精度高和可操作性好等优点,在生物产品分离领域中具有广阔的应用前景。
A new method of electrophoresis, that is multi-channel flow electrokinetic electrophoresis, was proposed. The microcomputer-controlled electrophoresis device was established and the influence of operating conditions on the separation efficiency of the device was investigated. Applying this technique, a continuous separation of bovine serum albumin (BSA) -batch of hemoglobin (HBB) was performed with 13.6 mg of BSA and 20.0 mg of HBB isolated from a 37.0 mg protein mixture containing BSA, HBB per hour. Application of this technology for purification of crude urokinase, its specific activity increased from 4000IU / mg to 4 × 10 ~ 4IU / mg or more, the yield of 10 × 10 ~ 4IU / h or more. This technique was combined with the (NH 4) SO 4 precipitation method to isolate 12.5 mg of high purity urokinase IgG from 15 ml of immunized rat serum at a yield of 2.1 mg / h. The above results prove that the multi-channel flow electrophoresis technology has the advantages of fast separation, high precision and good operability, and has broad application prospects in the field of biological product separation.