OPTIMIZING SEPARATION PARAMETERS IN CAPILLARY ISOELECTRIC-FOCUSING
OPTIMIZING SEPARATION PARAMETERS IN CAPILLARY ISOELECTRIC-FOCUSING
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DOI：
10.1016/0021-9673(91)80095-x
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发表时间：
1991-10-18
期刊：
影响因子：
--
通讯作者：
WEHR, T
中科院分区：
文献类型：
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作者：
ZHU, MD;RODRIGUEZ, R;WEHR, T
毛细管等电聚焦 (IEF) 已进行了多种改进，以改善蛋白质的分离和检测。 将碱性化合物掺入两性电解质混合物中，以在聚焦过程中阻断监测点和毛细管末端之间的部分，从而能够在动员过程中检测碱性蛋白质。 使用低等电点两性离子动员剂可提高 pH 梯度酸性区域的动员效率，从而改善酸性蛋白质的检测。 使用中性 pI 两性离子进行动员可选择性地动员中性和碱性蛋白质，并提高分辨率。 通过观察安装在热敏液晶上的玻璃毛细管中的有色蛋白质，可以确定沿着毛细管的热量产生模式以及盐对 IEF 过程的影响。 样品中盐的存在导致聚焦和动员时间较长。 在样品中加入非离子去污剂加两性电解质混合物可减少沉淀并提高 γ-球蛋白毛细管 IEF 的重现性。
Several modifications have been developed for capillary isoelectric focusing (IEF) to improve separation and detection of proteins. A basic compound was incorporated into the ampholyte mixture to block the segment between the monitor point and capillary end during focusing, enabling detection of basic proteins during mobilization. Use of a low-pI zwitterionic mobilization agent increased mobilization efficiency of the acidic region of the pH gradient, improving detection of acidic proteins. Mobilization with a neutral-pI zwitterion selectively mobilized neutral and basic proteins with improved resolution. Observation of colored proteins in glass capillaries mounted on thermosensitive liquid crystal was used to determine the heat generation patterns along the capillary and the effect of salt on the IEF process. The presence of salt in the sample resulted in long focusing and mobilization times. Incorporation of a non-ionic detergent in the sample plus ampholyte mixture reduced precipitation and improved reproducibility in capillary IEF of gamma-globulins.