三硝基苯磺酸法测定聚乙二醇化重组人生长激素的平均修饰度

目的:建立测定三种不同结构聚乙二醇化重组人生长激素(PEG-rhGH)平均修饰度的三硝基苯磺酸法(TNBS)。方法:采用基质辅助激光解吸附飞行时间质谱(MALDI-TOF)测定PEG-rhGH的分子量,求得理论平均修饰度。既而通过正交试验、游离PEG干扰等方法学考察,建立TNBS法。结果:从质谱法可知PEG-rhGH的理论修饰度为10%,TNBS法测得三种PEG-rhGH的修饰度分别为为10%、12%及10%,与理论值相符。结论:TNBS操作简单,不需要专门的仪器,可作为测定PEG-rhGH平均修饰度的方法。

Determination of Average Degree of Modification of Pegylated Recombinant Human Growth Hormone by Trinitrobenzene Sulfonic Acid

Objective: To determine the average extent of modification of PEG-rhGH by the established TNBS method. Methods: MALDI TOF mass spectrometry was used to detecte the molecular weight and the theoretical extent of modification of PEG-rhGH. TNBS method was established by the orthogonal experiment and the interference test of free PEG. Results: The actual modification ex- tent of three kinds of PEG-rhGHs were 10%, 12% and 10% respectively, and corresponding to the theoretical value of the modification extent it ...