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Protocol of capillary isoelectric focusing to separate extremely acidic and basic proteins
Author(s) -
Yang Chun,
Zhang Weibing,
Zhang Jie,
Duan Jicheng,
Zhang Yukui
Publication year - 2005
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200401793
Subject(s) - isoelectric focusing , chromatography , capillary electrophoresis , chemistry , isoelectric point , capillary action , lysozyme , immobilized ph gradient , electrophoresis , capillary electrochromatography , analytical chemistry (journal) , biochemistry , enzyme , materials science , composite material
A new set‐up was constructed for capillary isoelectric focusing (CIEF) involving a sampling capillary as a bypass fixed to the separation capillary. Sample solutions were subjected to a previously established pH gradient from the sample capillary. Besides performing conventional CIEF, the separation of ampholytic compounds with isoelectric points (p I s) beyond the pH gradient was carried out on this system. This method was termed as pH gradient driven electrophoresis (PGDE) and the basic mathematical expressions were derived to express the dynamic fundamentals. Proteins such as lysozyme, cytochrome C, and pepsin with p I s higher than 10 or below 3 were separated in a pH gradient provided by Pharmalyte (pH 3–10). Finally, this protocol convincingly exhibited its potential in the separation of a solution of chicken egg white.