Fractionation at pilot-plant scale of an haemoglobin hydrolysate by strong anionic exchange chromatography Application to the preparation of an amphiphilic peptide
✍ Scribed by Elisabeth Kosciarz; Florence Rofidal; Ida Dossou-Yovo; Dominique Vercaigne-Marko; Naïma Nedjar-Arroume; Pascal Dhulster; Didier Guillochon
- Publisher
- Wiley (John Wiley & Sons)
- Year
- 1998
- Tongue
- English
- Weight
- 275 KB
- Volume
- 71
- Category
- Article
- ISSN
- 0268-2575
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✦ Synopsis
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 cm3 Mono Q analytical column and progressively scaled-up to a 6 dm3 Q Sepharose Fast Flow column. For easy recovery of peptide and easy adjustment of conditions for Ðnal puriÐcation, a volatile bu †er, ethanolamine/HCl bu †er 20 mmol dm~3, pH 10É5, 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 puriÐcation 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.