TY  - JOUR
A1  - Mant, Colin T.
A1  - Chen, Yuxin
A1  - Yan, Zhe
A1  - Popa, Traian V.
A1  - Kovacs, James M.
A1  - Mills, Janine B.
A1  - Tripet, Brian P.
A1  - Hodges, Robert S.
T1  - HPLC analysis and purification of peptides
T2  - Methods in molecular biology
N2  - High-performance liquid chromatography (HPLC) has proved extremely versatile over the past 25 yr for the isolation and punfication 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 peptideslproteins by sizeexclusion chromatography and monitoring the hydrophilicitykydrophobicity of amphipathic cr-helical peptides, a vital precursor Tor 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. Key Words: Peptides; proteins; size-exclusion chromatography (SEC); anion-exchange chromatography (AEX); cation-exchange chromatography (CEX); mixed-mode hydrophilic interaction chromatography (HIL1C)/cation-exchange chromatography (CEX); reversed-phase high-performance liquid chromatography (RP-HPLC); preparative RP-HPLC of peptides and proteins; amino acid side-chain hydrophilicitylhydrophobicity coefficients; amino acid U-helical propensity values; amino acid side-chain stability coefficients
KW  - proteins
KW  - size-exclusion chromatography (SEC)
KW  - anion-exchange chromatography (AEX)
KW  - cation-exchange chromatography (CEX)
Y1  - 2007
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/9627
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30-1107691
SN  - 1064-3745
N1  - Signatur: SR 2894
IS  - 386
SP  - 3
EP  - 55
ER  -