反相高效液相层析在糖化白蛋白肽段分离纯化中的应用  

The Application of RP HPLC in the Analysis and Purification of an Immunogen:Glycoalbumin Fragment.

作　　者：梁逊[1,2] 李顺子[1,2] 李文兰 

机构地区：[1]南开大学化学系 [2]南开大学吸附分离功能高分子材料国家重点实验室

出　　处：《生物化学与生物物理进展》1999年第1期75-78,共4页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金

摘　　要：采用一种简单的“甲醇水三氟醋酸”作为洗脱体系的反相高效液相层析（简称ＲＰＨＰＬＣ）对通过固相合成方法合成的糖化白蛋白肽段进行了分析鉴定和分离纯化．使用酸敏性ＰＥＧ载体，Ｆｍｏｃ保护化学法合成了白蛋白八肽ＮＨ２ＬｙｓＧｌｎＴｈｒＡｌａＬｅｕＴｙｒＴｙｒＣｙｓＣＯＯＨ．对其Ｎ端Ｌｙｓ进行糖化反应后，经ＳｅｐｈａｄｅｘＧ１０柱色谱纯化后，通过ＲＰＨＰＬＣ分析，证明得到了糖化八肽的单一峰．使用Ｍｅｒｒｉｆｉｅｌｄ树脂，Ｂｏｃ保护化学法合成了白蛋白七肽ＮＨ２ＧｌｎＴｈｒＡｌａＬｅｕＴｙｒＴｙｒＣｙｓＣＯＯＨ．通过ＲＰＨＰＬＣ半制备分离提纯后，得到了所需的肽段．对ＮαＢｏｃＬｙｓ的ε氨基进行糖化反应后，经ＲＰＨＰＬＣ分析，证明得到了比较纯的糖化赖氨酸，与纯化后的白蛋白七肽偶联后，通过ＲＰＨＰＬＣ分析，得到了偶联产物———糖化八肽的单一峰．RP HPLC method with a simple “MeOH H 2O TFA” elution system is applied in purification and analysis of human glycoalbumin fragment including the region of lysine 525, which was synthesized by different methods. The peptide fragment of albumin KQTALYYC prepared by acid labile PEG resin was glycosylated and purified by gel filtration and analysed by RP HPLC. A single peak demonstrated that the glycosylation of the peptide was successful . Heptapeptide fragment of albumin QTALYYC was synthesized by chloromethyl resin according to the Boc chemistry strategy. The pure heptapeptide was obtained by semi preparative RP HPLC. Glycosylated lysine was purified by gel filtration and ascertained by RP HPLC. The heptapeptide was coupled with glycosylated lysine. RP HPLC of the product showed that the glycopeptide is qualified as a semiantigen.

关 键 词：糖化白蛋白 糖化赖氨酸 RP-HPLC 分离 

分 类 号：Q513.203[生物学—生物化学]