人唾液蛋白的高效疏水色谱分离纯化

目的　探讨高效疏水色谱分离、纯化人全唾液蛋白、腮腺液蛋白及颌 /舌下腺液蛋白的最佳分离条件和临床意义。方法　将收集的唾液样品经低温离心 ,过滤 ,冻干 ,低温保存备用。以不同浓度的 (NH4 ) 2 SO4 、KH2 PO4 溶液分别为A、B流动相 ,梯度分离 ,收集各个组分峰 ,通过生化方法对蛋白进行定性。结果　全唾液蛋白被分离为 12个峰组分 ,腮腺液被分离为 6个峰组分 ,颌 /舌下腺液被分为 8个峰组分。经统计学处理 ,平均出峰时间标准差小 ,个体间重复性好。目前能初步定性的唾液蛋白为富含脯氨酸蛋白、半胱氨酸蛋白、富酪蛋白和富组蛋白及α 淀粉酶。结论　疏水色谱可用于人全唾液蛋白、腮腺液蛋白及颌 /舌下腺液蛋白的分离

Separation and purification of human salivary protein byhigh-performance hydrophobicity chromatography

Objective To separate and purify human whole salivary protein, parotid salivary protein and submandibular/sublingual salivary protein by HPHIC to discuss the best separation condition and its clinical significance. Methods Each kind of salivary sample was collected, centrifugated and lyophilized at low temperature and stored for use. After the gradient was separated by different concentrations of (NH 4) 2SO 4, KH 2PO 4 as A and B phase, each spike was collected and qualified by biochemical techniques. Results There were 12, 6 and 8 protein components in the whole salivary, parotid salivary and submandibular/sublingual salivary, respectively, with minimal standard error and good duplication. Five kinds of protein could be qualified, such as proline-rich protein, statherin, cystatin S, α-amylase and histone-rich protein. Conclusion HPHIC can be used to separate human whole salivary protein, parotid salivary potein and submandibular/sublingual salivary protein.