摘要目的：使用等电聚焦电泳（IEF）技术准确地测定甘精胰岛素的等电点。方法：甘精胰岛素用双蒸水溶解（可加微量醋酸溶液助溶），利用真空冻干机进行冷冻干燥。将冷冻干燥后的供试品用含pH 3。0-10。0载体两性电解质的上样缓冲液溶解，进行等电聚焦电泳，测量电泳后的图谱中等电点标准蛋白（pI Marker）距阳极的泳动距离，绘制标准曲线, 计算甘精胰岛素的等电点值。结果表明：重组甘精胰岛素供试品的等电点与重组甘精胰岛素标准品的等电点一致，pI均约为6。7，标准曲线线形回归方程为y=-0。797x+9。7658，相关系数r 达0。9956。这说明使用等电聚焦电泳测定甘精胰岛素的等电点重复性好，精确度较高，方法准确，为检测胰岛素以及胰岛素类似物的等电点提供参考，为甘精胰岛素的生产提供理化指标。75032

该论文有图4幅，参考文献11篇。

毕业论文关键词：甘精胰岛素  等点聚焦电泳  等电点

Determination of Isoelectric Point of Insulin Glargine

Abstract Objective: To detect the isoelectric point of insulin glargine by using the isoelectric focusing (IEF) technology。 Methods: the insulin glargine was dissolved with the doudle distilled water (could add a trace of acetic acid) , and it was frozen and dried by freeze-drying apparatus。 The frozen and dried insulin glargine was solubilized by the loading buffer containing pH3。0-10。0 carrier and was performed by the isoelectric focusing electrophoresis。 Then, we can measure the swimming distance from the anode of pI Marker in the map of electrophoresis。 Finally the isoelectric point of insulin glargine was calculated by the standard curve。 Results: The pI of insulin glargine was about 6。7, and this was coincident with the standed insulin glargine。 The linear regression equation of curve was y=-0。797x+9。7658, the correlation coefficient r was 0。9956。 And this indicated that the isoelectric focusing electrophoresis was an accurate method for detection of the isoelectric point of insulin glargine。 It has good reproducibility and high accuracy。 This article could provide a good reference for the determination the pI of other insulins and insulin analogues ,and provide physical and chemical indicators for the production of insulin glargine。

This paper has four pictures and eleven references。

Key Words: insulin glargine  isoelectric focusing electrophoresis  isoelectric point

目  录

摘  要 I

Abstract II

目  录 III

1 绪论 1

2 材料与方法 2

2。1 供试材料 2

2。2 主要设备 2

2。3 主要试剂 3

2。4 试剂配置 3

2。5 等电聚焦电泳测定甘精胰岛素等电点 3

3 结果与分析 5

3。1 甘精胰岛素等电点测定图谱 5

3。2 pI与迁移距离间的线性回归方程 7

4 讨论 7

5 结论 8

参考文献 9

致谢 10

图清单

图序号 图名称 页码

图3-1 甘精胰岛素样品1与BSA的等电聚焦电泳图谱