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HPLC analysis and purification of peptides.
- 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.
- Subjects :
- Amino Acid Sequence
Amino Acids chemistry
Animals
Antimicrobial Cationic Peptides chemistry
Antimicrobial Cationic Peptides isolation & purification
Chromatography, High Pressure Liquid instrumentation
Chromatography, High Pressure Liquid standards
Chromatography, Ion Exchange methods
Drug Stability
Electrophoresis, Capillary methods
Hydrophobic and Hydrophilic Interactions
Molecular Biology methods
Molecular Sequence Data
Oligopeptides chemistry
Oligopeptides isolation & purification
Peptides chemistry
Peptides standards
Protein Conformation
Protein Structure, Secondary
Recombinant Proteins chemistry
Recombinant Proteins isolation & purification
Reference Standards
Temperature
Chromatography, High Pressure Liquid methods
Peptides isolation & purification
Subjects
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