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Improved purification process of β- and α-trypsin isoforms by ion-exchange chromatography BABT
Santos,Alexandre Martins Costa; Oliveira,Jamil Silvano de; Bittar,Eustáquio Resende; Silva,Anderson Lourenço da; Guia,Marcos Luiz dos Mares; Bemquerer,Marcelo Porto; Santoro,Marcelo Matos.
The purpose of this work was to improve the separation and yield of pure β- and α-trypsin isoforms by ion-exchange chromatography and to characterize some physical-chemical properties of these isoforms. Purification of trypsin isoforms was performed by ion-exchange chromatography in 0.1 mol/L tris-HC buffer, pH 7.10 at 4ºC. The sample loading, salt concentration, flow rate and pH of mobile phase were varied to determine their effects on the resolution of the separation. The resolution was optimized mainly between β- and α-trypsin. Pure isoforms were obtained by chromatographying 100 mg of commercial trypsin during seven days, yielding 51 mg of high purity β-trypsin and 13 mg of α-trypsin partially pure, with small amounts of contaminating of ψ-trypsin....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Trypsin isoforms; Resolution; Ion-exchange chromatography; Purification; Specific activity; Mass spectrometry.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000400009
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