title: Two novel solvent system compositions for protected synthetic peptide purification by centrifugal partition chromatography.
abstract: Protected synthetic peptide intermediates are often hydrophobic and not soluble in most common solvents. They are thus difficult to purify by preparative reversed-phase high-performance liquid chromatography (RP-HPLC), usually used for industrial production. It is then challenging to develop alternative chromatographic purification processes. Support-free liquid-liquid chromatographic techniques, including both hydrostatic (centrifugal partition chromatography or CPC) and hydrodynamic (counter-current chromatography or CCC) devices, are mainly involved in phytochemical studies but have also been applied to synthetic peptide purification. In this framework, two new biphasic solvent system compositions covering a wide range of polarity were developed to overcome solubility problems mentioned above. The new systems composed of heptane/tetrahydrofuran/acetonitrile/dimethylsulfoxide/water and heptane/methyl-tetrahydrofuran/N-methylpyrrolidone/water were efficiently used for the CPC purification of a 39-mer protected exenatide (Byetta®) and a 8-mer protected peptide intermediate of bivalirudin (Angiox®) synthesis. Phase compositions of the different biphasic solvent systems were determined by (1)H nuclear magnetic resonance. Physico-chemical properties including viscosity, density and interfacial tension of these biphasic systems are also described.
authors.affiliation: ICMR UMR CNRS 7312, IFR 53 Interactions Cellules Microenvironnement, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, B.P. 1039, 51687 Reims cedex 2, France.
authors.forename: Nassima
authors.initials: N
authors.lastname: Amarouche
authors.affiliation: Lonza AG, Valais Works, Rottenstrasse 6, 3930 Visp, Switzerland.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Giraud
authors.affiliation: Lonza AG, Valais Works, Rottenstrasse 6, 3930 Visp, Switzerland.
authors.forename: Luciano
authors.initials: L
authors.lastname: Forni
authors.affiliation: Lonza AG, Valais Works, Rottenstrasse 6, 3930 Visp, Switzerland.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Butte
authors.affiliation: ICMR UMR CNRS 7312, IFR 53 Interactions Cellules Microenvironnement, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, B.P. 1039, 51687 Reims cedex 2, France.
authors.forename: F
authors.initials: F
authors.lastname: Edwards
authors.affiliation: ICMR UMR CNRS 7312, IFR 53 Interactions Cellules Microenvironnement, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, B.P. 1039, 51687 Reims cedex 2, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Borie
authors.affiliation: ICMR UMR CNRS 7312, IFR 53 Interactions Cellules Microenvironnement, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, B.P. 1039, 51687 Reims cedex 2, France. Electronic address: jh.renault@univ-reims.fr.
authors.forename: Jean-Hugues
authors.initials: JH
authors.lastname: Renault
mesh_terms: D003377:Countercurrent Distribution / Q000379:methods; D000077270:Exenatide; D006629:Hirudins / Q000302:isolation & purification*; D009682:Magnetic Resonance Spectroscopy; D010446:Peptide Fragments / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012995:Solubility; D012997:Solvents / Q000737:chemistry*; D014688:Venoms / Q000302:isolation & purification*
chemical_list: D006629:Hirudins; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D012997:Solvents; D014688:Venoms; D000077270:Exenatide; C074619:bivalirudin
keywords: Biphasic solvent systems; centrifugal partition chromatography (CPC); preparative chromatography; protected peptides