Boss H J, Rohde M F, Rush R S
Amgen, Thousand Oaks, CA, USA.
Pept Res. 1996 Jul-Aug;9(4):203-9.
This papers addresses many of the optimization parameters necessary to convert from high resolution capillary electrophoresis (CE) analytical separation parameters to automated, micropreparative multiple fraction collection using software-controlled, interrupted applied voltage. Optimization of two parameters are crucial: 1) preparative sample loading and 2) the determination of peak collection windows. Factors affecting sample loading volume are discussed, such as capillary inner diameters, sample temperatures and sample injection times. Peak collection windows have been determined experimentally and offer an advantage to windows calculated using a linear mobility relationship, especially for long run times, high current levels, and multiple voltage ramping required for multiple fraction collection. Reinjection analysis of both non-glycopeptides and glycopeptides are examined, and clearly indicate peak mobility can be employed for identifying the collected peptides. Difficulties associated with quantitation of the collected peaks by CE are described and appear to be predominantly associated with sample matrix effects.
本文论述了从高分辨率毛细管电泳（CE）分析分离参数转换为使用软件控制的间断施加电压进行自动化微量制备多馏分收集所需的许多优化参数。两个参数的优化至关重要：1）制备性进样；2）峰收集窗口的确定。讨论了影响进样体积的因素，如毛细管内径、样品温度和进样时间。峰收集窗口已通过实验确定，与使用线性迁移率关系计算的窗口相比具有优势，特别是对于长时间运行、高电流水平以及多馏分收集所需的多次电压梯度变化。研究了非糖肽和糖肽的再进样分析，明确表明峰迁移率可用于鉴定收集到的肽段。描述了通过CE对收集到的峰进行定量时遇到的困难，这些困难似乎主要与样品基质效应有关。
