摘要
分别采用基质辅助激光解吸离子化-飞行时间质谱法(MALDI-TOF MS)、Tricine-SDS-聚丙烯酰胺凝胶电泳法(Tricine-SDS-PAGE)和分子排阻色谱法(SEC)测定重组人甲状旁腺素1-34的相对分子质量,为制定该品种的质量标准提供依据。MALDI-TOF/MS测定基质溶液为α-氰基-4-羟基肉桂酸(a-cyano-4-hydroxy-cinnamic acid,CCA);Tricine-SDS-PAGE电泳采用Tris-tricine电极缓冲系统;SEC测定色谱柱为TSK-GELG2 000 SWXL(300mm×7.8mm,5μm);流动相为三氟乙酸-乙腈-水(0.05∶20∶80);检测波长为215nm;流速为0.5 mL.min-1。结果:质谱法测定结果准确,实验值与理论值一致,误差±1;Tricine-SDS-PAGE测定结果与理论值基本一致,误差小于10%;但SEC法测定结果与理论值偏差较大。质谱法适用于产品的质量控制。
To deter mine the relative molecular weight of Recombinant Human Parathyriod Hormone 1- 34（rhPTH1-34）by matrix assisted laser desorptiny ionization time of flight Mass Spectrometry （MALDI-TOFMS） ,Tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis（Tricine-SDS-PAGE）and size exclusive chromatography （SEC） respectively to provide the basis for the specification of rhPTH1- 34. The matrix for MALDI-TOF MS was α-cyano-4-hydroxycinnamic acid（CCA）. The electrode buffer was Tris-Tricine system. SEC column was TSK-GEL G2000 SWXL （300mm×7. 8mm,5 μm）. The mobile phase was trifluoroacetic - acetonitrile - water （0. 05 ：20 ： 80）. The detection wavelength was 215nm. The flow rate was 0. 5 mL · min^-1. The results of MALDI-TOF MS are accurate. The measured value is identical with the theoretical value with,error of ±1. The measured value of Tricine-SDS- PAGE is the same as the theoretical value with a deviation of less than 10%. The results of SEC are different from the theoretical value. MS is suitable for the quality control of the products.
出处
《药物生物技术》
CAS
CSCD
2006年第4期286-289,共4页
Pharmaceutical Biotechnology
