HPLC Methods for Purification of Antimicrobial Peptides
The advent of high performance liquid chromatography (HPLC) has greatly accelerated the discovery, purification, and characterization of antimicrobial peptides. Virtually every modern study of an antimicrobial peptide includes or was preceded by a description of its purification. The increased pace of peptide discovery and characterization has resulted from the development of sophisticated column and solvent delivery technology over the last four decades. Interestingly, the modern methods described here derive directly from standard open-column (“low performance”) chromatographic modalities. Therefore, it is not surprising that virtually every method used in traditional column chromatography has been adapted to high performance methods. These include gel filtration, ion-exchange, and reversed-phase chromatography methods described in this chapter.
KeywordsHigh Performance Liquid Chromatography Filtration Acetonitrile Helium Hydrocarbon
- 8.Selsted, M. E., Tang, Y.-Q., Morris, W. L., McGuire, P. A., Novotny, M. J., Smith, W., Henschen, A. H., and Cullor, J. S. (1993) Purification, primary structures, and antibacterial activities of β-defensins, a new family of antimicrobial peptides from bovine neutrophils. J. Biol. Chem. 268, 6641–6648.PubMedGoogle Scholar
- 28.Anderson, J. K. and Mole, J. E. (1983) Adaptation of reverse-phase high-performance liquid chromatography for the isolation and sequence analysis of peptides from plasma amyloid p-component, in High-Performance Liquid Chromatography of Proteins and Peptides (Hearn, M. T. W., Regnier, F. E., and Wehr, C. T., eds.), Academic, New York, pp. 29–37.Google Scholar
- 29.Selsted, M. E. (1993) Investigational approaches for studying the structures and biological functions of myeloid antimicrobial peptides, in Genetic Engineering: Principles and Methods (Setlow, J. K., ed.), Plenum, New York, pp 131–147.Google Scholar
- 30.Bennett, H. P. J., Browne, C. A., and Solomon, S (1983) α-N-Acetyl-β-Endorphin1-26 from the neurointermediary lobe of the rat pituitary isolation, purification, and characterization by high-performance liquid chromatography, in High-Performance Liquid Chromatography of Proteins and Peptides (Hearn, M. T. W., Regnier, F. E., and Wehr, C. T., eds.), Academic, New York, p 253–261.Google Scholar