二烯丙基聚乙二醇20M交联键合型毛细管电泳柱的制备及其评价

Preparation of Cross-linked and Surface Bonded Diallyl-Polyethlene Glycol 20M Coated Column for Capillary Electrophoresis of Basic Proteins

本文将聚乙二醇20M改性为二烯丙基聚乙二醇20M，使其更容易在毛细管壁进行，交联和键合，采用超动态涂柱技术，实现了快速制备二烯丙基聚乙二醇20M交联缝合型毛细管电泳柱。该制柱方法具有简单，速度快，制柱重复性高的特点。用四种碱性蛋白对所制的柱进行评价，平均柱效可达5×105理论塔板数／米。每次运行之间（N=6），天与天之间（N=3），以及柱与柱之间（N=3）的迁移时间的标准偏差（RSD％）在0．2％～0．8％之间。对所制的柱进行上百次的进样，保存3个月后，柱效平均下降约20％，表明用该方法所制的柱具有良好的稳定性。

A simpie and rapid method for the preparation of cross-linked and surface bonded diallyl-polyethylene glycol (PEG) 20M coated capillary used for the separation of basic proteins by capillary electrophoresis was described. In one approach, the two hydroxy groups at the both ends of PEG 20M molecule were chemically modified by allyl bromide through williamson reaction; in second approach, the capillary was silanized with villyldimethoxysilane ; in third approach, the capillary was dynamically coated with the solution containing diallyl-PEG 20M (5%,m/V) and dicumyl pcroxide (5% ) in mixture solvent of isopropane ether and methylene chloride (V/V= 1: 1 ). After this, thermal treatment was preformed to cross-link and immobilize diallyl-PEG 20M on to the capillary surface. Four basic proteins were used for evaluating the coated capillary, the average separation efficiency of 5×105 plate/m was obtained. Good reproducibilities (RSD%<1) of migration times from run to run, day to day, column to column were obtained. After hundreds of injections, no loss of efficiency was observed.