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寡聚化磷脂双层作为毛细管电泳中的半永久性涂层

Oligomerized phospholipid bilayers as semipermanent coatings in capillary electrophoresis.

作者信息

Wang Chuanzhong, Lucy Charles A

机构信息

Department of Chemistry, University of Alberta, Gunning/Lemieux Chemistry Centre, Edmonton, Alberta, T6G 2G2 Canada.

出版信息

Anal Chem. 2005 Apr 1;77(7):2015-21. doi: 10.1021/ac0489622.

Abstract

Double-chained surfactants form semipermanent coatings that prevent protein adsorption in capillary electrophoresis (CE). To make such coatings more permanent, vesicles of the unsaturated phospholipid 1,2-dioleoyl-sn-glycero-3-phosphocholine were prepared and subjected to free-radical-initiated polymerization, both inside the capillary and in free solution. The latter generated oligomers of 2-5 units based on ESI-TOF MS, and formed the more stable coating in CE. Rinsing the capillary with a solution of the ex situ oligomerized DOPC suppressed EOF (0.8 x 10(-)(8) m(2)/V.s) for more than 20 h, whereas in situ oligomerized electroosmotic flow (EOF) suppressed the EOF for only 10 h. Mixtures of anionic and cationic proteins were separated under neutral pH and low ionic strength buffer with efficiencies of 480,000-930,000 plates/m and recoveries of 75-99%.

摘要

双链表面活性剂形成半永久性涂层,可防止蛋白质在毛细管电泳(CE)中吸附。为使此类涂层更持久,制备了不饱和磷脂1,2 - 二油酰 - sn - 甘油 - 3 - 磷酸胆碱的囊泡,并在毛细管内部和自由溶液中进行自由基引发的聚合反应。基于电喷雾电离飞行时间质谱(ESI - TOF MS),后者生成了2 - 5个单元的低聚物,并在毛细管电泳中形成了更稳定的涂层。用异位低聚的二油酰磷脂酰胆碱(DOPC)溶液冲洗毛细管可在超过20小时内抑制电渗流(EOF,0.8×10⁻⁸ m²/V·s),而原位低聚的电渗流仅能抑制电渗流10小时。在中性pH和低离子强度缓冲液条件下,阴离子和阳离子蛋白质混合物的分离效率为480,000 - 930,000理论塔板数/米,回收率为75 - 99%。

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