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Fractionation at pilot‐plant scale of an haemoglobin hydrolysate by strong anionic exchange chromatography Application to the preparation of an amphiphilic peptide
Author(s) -
Kosciarz Elisabeth,
Rofidal Florence,
DossouYovo Ida,
VercaigneMarko Dominique,
NedjarArroume Naïma,
Dhulster Pascal,
Guillochon Didier
Publication year - 1998
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/(sici)1097-4660(199801)71:1<35::aid-jctb779>3.0.co;2-l
Subject(s) - chromatography , chemistry , hydrolysate , fractionation , peptide , amphiphile , ion exchange , ion chromatography , electrodialysis , membrane , ion , biochemistry , organic chemistry , copolymer , hydrolysis , polymer
The development and the scale‐up of high performance anion chromatography to obtain 1 milligram to 1 gram yields of a peptide fraction from a complex peptic haemoglobin hydrolysate is described here. The chromatographic conditions were developed using a 1 cm 3 Mono Q analytical column and progressively scaled‐up to a 6 dm 3 Q Sepharose Fast Flow column. For easy recovery of peptide and easy adjustment of conditions for final purification, a volatile buffer, ethanolamine/HCl buffer 20 mmol dm −3 , pH 10·5, was employed; desalting was carried out by a pilot‐plant scale electrodialysis which permitted the elimination of 99% NaCl without important loss of peptide (less than 15%). A combination of these techniques with reverse phase HPLC proved a useful strategy for fractionation of a complex peptide mixture and enabled pure peptides to be obtained in sufficient quantities for further analyses and biological tests. The example of preparation and purification of an amphiphilic peptide is described. Its ability to solubilize an insoluble photosensitizer, protoporphyrin IX, was determined in order to study its utilization as a carrier for photochemotherapy. © 1998 SCI.