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目的:对重组人甲状旁腺素rh-PTH(1-34)结构类似物(以下简称rhPTH′)的纯度进行质量控制,为其生产工艺提供实验室研究依据。方法:采用聚丙烯酰胺小肽凝胶电泳(SDS-PAGE)、等电聚焦电泳(IFE)、氨基酸组成分析和反相高效液相色谱法(RP-HPLC)对制备的rhPTH′的纯度等进行质量鉴定,并配合质谱法测定样品的精确相对分子质量。结果:经鉴定酸水解后目的蛋白纯度为89.2%;离子交换层析后样品纯度经电泳和HPLC鉴定分别为100%和94.9%;样品等电点为9.05,与理论值基本一致;氨基酸组成与理论组成相同,质谱鉴定相对分子质量为4663.76,与理论值4664.01相差0.25。结论:用现有生产和纯化工艺制备的rhPTH′具有较高的纯度,质量符合要求。
OBJECTIVE: To control the purity of recombinant human parathyroid hormone rh-PTH (1-34) (hereinafter referred to as rhPTH ’), and to provide a laboratory basis for its production technology. Methods: Purity of rhPTH ’prepared by polyacrylamide gel electrophoresis (SDS-PAGE), isoelectric focusing (IFE), amino acid composition analysis and reverse phase high performance liquid chromatography (RP-HPLC) Quality identification, and with the mass spectrometry determination of the exact molecular weight of the sample. Results: The purity of the target protein after acid hydrolysis was 89.2%. The purity of the sample after ion exchange chromatography was 100% and 94.9% respectively by electrophoresis and HPLC. The isoelectric point of the sample was 9.05, which was consistent with the theoretical value. The same theoretical composition, mass spectrometry identified relative molecular mass of 4663.76, with the theoretical value of 4664.01 difference of 0.25. Conclusion: rhPTH ’prepared by current production and purification process has high purity and meets the requirements of quality.