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Application of free‐flow electrophoresis to the purification of trichosanthin from a crude product of acetone fractional precipitation
Author(s) -
Song JinFang,
Liu Tao,
Shen Xin,
Wu Gaode,
Xia Qichang
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150190708
Subject(s) - chromatography , acetone , trichosanthin , precipitation , chemistry , electrophoresis , biochemistry , physics , meteorology
Abstract The application of free‐flow electrophoresis (FFE) to the purification of trichosanthin (TCS) from a crude product of acetone fractional precipitation was investigated. An electrophoresis technique, combining field step electrophoresis (FSE) and zone electrophoresis (ZE) to a one‐step procedure, was optimized until a satisfactory purification factor (1.35), high resolution, and purity (>99%) were achieved. Testing several separation buffer systems revealed that a throughput of 14.2 mg/h can be obtained when the very basic TCS (p I 10.1) was dissolved and electrophoresed in a phosphate buffer system of pH 4. The purity of electrophoresed trichosanthin was proved by a variety of analytical methods, such as sodium dodecyl sulfate (SDS)‐gel electrophoresis, capillary isoelectric focusing (CIEF), and sequencing of N ‐ and C ‐termini. The high purity and large throughput achieved at low cost by using FFE indicates that this method can be employed for TCS purification.