重组人粒细胞巨噬细胞集落刺激因子的生物质谱和毛细管电泳法  

CHARACTERIZATION OF RECOMBINANT HUMAN GRANULOCYTE MACROPHAGE COLONY STIMULATING FACTORS (rHuGM CSF) BY BIOCHEMICAL MASS SPECTROMETRY AND CAPILLARY ELECTROPHORESIS

作　　者：周国华[1,2,3] 罗国安[1,2,3] 张晓丹 曹亚澄[1,2,3] 孙国庆 

机构地区：[1]清华大学化学系 [2]南京军区药品检验所 [3]中国科学院南京土壤研究所

出　　处：《药学学报》1999年第8期621-626,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金;国家新药博士基金

摘　　要：目的：为准确分析重组人粒细胞巨噬细胞集落刺激因子（ｒＨｕＧＭＣＳＦ）半成品及其制剂的纯度和表达的正确性。方法：用毛细管区带电泳测定了２批ｒＨｕＧＭＣＳＦ半成品和３批ｒＨｕＧＭＣＳＦ注射液的纯度。用基体辅助激光解吸飞行时间质谱和电喷雾质谱测定了２批ｒＨｕＧＭＣＳＦ半成品的分子量。用毛细管等电聚集法测定了ｒＨｕＧＭＣＳＦ半成品的等电点。结果：２批半成品中１批含有少量二聚体，且含有４个不同等电点的组分，另１批表达不正确；２批成品均存在大量杂质。结论：发现有些ｒＨｕＧＭＣＳＦ样品有生物学活性，但表达不正确，纯度也不高；各分析方法是互补的，不能相互替代。AIM: To accurately determine the purity and correctness of expression for the bulk samples and preparations of rHuGM CSF with biological activity. METHODS: The purities of two bulk samples and three preparations of rHuGM CSF were determined by capillary zone electrophoresis (CZE), molecular weights of two rHuGM CSF bulk samples by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI TOF/MS) and electrospray ionization mass spectrometry (ESI/MS), and isoelectric points of rHuGM CSF bulk sample by capillary isoelectric focusing (CIEF). RESULTS: One of the two bulk samples was expressed incorrectly, and the other contained a small amount of dimer, which composed of four components with different isoelectric points. There were large amounts of impurities in the two rHuGM CSF preparations. CONCLUSION: The sample of rHuGM CSF with biologic activity may be expressed incorrectly or contained impurities. The instrumental methods applied in this paper are complementary and we can not use one to replace the other.

关 键 词：RHUGM-CSF 毛细管电泳法 生物质谱 

分 类 号：Q517.03[生物学—生物化学] R963[医药卫生—微生物与生化药学]