UNIT 10.13 HPLC of Peptides and Proteins: Standard Operating Conditions
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Boysen, R. I. and Hearn, M. T. 2001. HPLC of Peptides and Proteins: Standard Operating Conditions. Current Protocols in Molecular Biology. 54:V:10.13:10.13.1–10.13.32.
- Published Online: 1 MAY 2001
- Published Print: APR 2001
The standard operating conditions for the eight basic modes of HPLC are presented in this unit. They include: size-exclusion chromatography (HP-SEC), ion-exchange chromatography (HP-IEX), normal phase chromatography (HP-NPC), hydrophobic interaction chromatography (HP-HIC), reversed-phase chromatography (RP-HPLC), hydrophilic interaction chromatography (HP-HILIC), immobilized metal ion affinity chromatography (HP-IMAC), and biospecific/biomimetic affinity chromatography (HP-BAC). In addition, some subsets of these chromatographic modes, e.g., mixed mode chromatography (HP-MMC), charge transfer chromatography (HP-CTC), or ligand-exchange chromatography (HP-LEC) are described. Procedures for multimodal column switching are also included, as are guidelines for a systematic approach to method development. Example separations help illustrate the procedures.The standard operating conditions for the eight basic modes of HPLC are presented in this unit.