1. Two novel solvent system compositions for protected synthetic peptide purification by centrifugal partition chromatography.
- Author
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Amarouche N, Giraud M, Forni L, Butte A, Edwards F, Borie N, and Renault JH
- Subjects
- Countercurrent Distribution methods, Exenatide, Magnetic Resonance Spectroscopy, Recombinant Proteins isolation & purification, Solubility, Hirudins isolation & purification, Peptide Fragments isolation & purification, Peptides isolation & purification, Solvents chemistry, Venoms isolation & purification
- 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., (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Published
- 2014
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