重组葡激酶脂质体的制备、活性及包封率测定方法的研究

目的:研究重组葡激酶脂质体的制备、活性及其包封率的测定方法,为重组葡激酶脂质体的研究开发提供实验数据。方法:薄膜分散法制备重组葡激酶脂质体并用溶圈法测定其活性,分别采用超速离心和直接点样2种方法测定重组葡激酶脂质体的包封率。结果:溶圈法测定重组葡激酶活性的板内精密度的 RSD 为8.4%(n=5),板间精密度的 RSD 为12.4%(n=5)。直接点样法测定所制备的3批重组葡激酶脂质体包封率分别为54.36%,51.67%,54.67%;离心法测定所制备的3批重组葡激酶脂质体包封率分别为71.75%,67.43%,68.46%。所制备的重组葡激酶脂质体表面药物吸附量为15.65%。结论:溶圈法可用于重组葡激酶脂质体活性的测定;离心法测定的重组葡激酶脂质体包封率包括脂质体表面吸附药量;离心法与直接点样法测定包封率的差值为脂质体表面吸附药物量。

Study on preparation of recombinant staphylokinase liposome and determination of its activity and entrapment efficiency

Objective:To prepare recombinant staphylokinase(r-Sak)liposome and develop a methods to determi- nate its activity and entrapment efficiency for providing the experimental basis in the study and development of r - Sak liposome preparation.Methods:The liposomes containing r-Sak were prepared by film dispersion method and their fibrinolytic activities were determined bylyric circlemethod.Entrapment efficiency was respectively meas- ured by ultracentrifugation spotting method and directly spotting method.Results:The RSDs(n=5) of intra-plates and inter-plates for assay of fibrinolytic activity with lyric circle method were 8.4% and 12.4% respectively.The entrapment efficiencies of three batches of r-Sak liposomes were 54.36%,51.67% and 54.67% respectively u- sing directly spotting method and 71.75%,67.43% and 68.46% respectively using ultracentrifugation spotting as- say method.The adsorption percentage of r-Sak on the surface of liposomes was 15.65%.Conclusions:Lyric cir- cle method can be used to assay the activities of r-Sak liposomes,and the entrapment efficiencies of r-Sak in li- posomes determined by using ultracentrifugation spotting assay contained the amounts of r-Sak adsorbed on surface of liposome.The difference of two entrapment efficiencies obtained respectively from using ultracentrifugation spot- ting assay and directly spotting method was the amounts of r-Sak adsorbed on surface of liposome.