Back to Search Start Over

HPLC analysis and purification of peptides.

Authors :
Mant CT
Chen Y
Yan Z
Popa TV
Kovacs JM
Mills JB
Tripet BP
Hodges RS
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2007; Vol. 386, pp. 3-55.
Publication Year :
2007

Abstract

High-performance liquid chromatography (HPLC) has proved extremely versatile over the past 25 yr for the isolation and purification of peptides varying widely in their sources, quantity and complexity. This article covers the major modes of HPLC utilized for peptides (size-exclusion, ion-exchange, and reversed-phase), as well as demonstrating the potential of a novel mixed-mode hydrophilic interaction/cation-exchange approach developed in this laboratory. In addition to the value of these HPLC modes for peptide separations, the value of various HPLC techniques for structural characterization of peptides and proteins will be addressed, e.g., assessment of oligomerization state of peptides/proteins by size-exclusion chromatography and monitoring the hydrophilicity/hydrophobicity of amphipathic alpha-helical peptides, a vital precursor for the development of novel antimicrobial peptides. The value of capillary electrophoresis for peptide separations is also demonstrated. Preparative reversed-phase chromatography purification protocols for sample loads of up to 200 mg on analytical columns and instrumentation are introduced for both peptides and recombinant proteins.

Details

Language :
English
ISSN :
1064-3745
Volume :
386
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
18604941
Full Text :
https://doi.org/10.1007/978-1-59745-430-8_1