Unit

UNIT 10.13 HPLC of Peptides and Proteins: Standard Operating Conditions

  1. Reinhard I. Boysen,
  2. Milton T.W. Hearn

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1013s54

Current Protocols in Molecular Biology

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.

Author Information

  1. Monash University, Victoria, Australia

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 2001

Abstract

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.