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Microisolation of Biologically Active Polypeptides by Reverse-Phase Liquid Chromatography

  • Petro E. Petrides
Protocol
Part of the Biological Methods book series (BM)

Abstract

The development of new, high-sensitivity methods in protein chemistry (microisolation, amino acid microanalysis, and microse-quencing) has permitted the characterization of a large number of previously unknown polypeptides with regulatory functions. After decades of research devoted to the metabolism of the individual cell, this new methodology has sparked investigation into the presence and functional significance of polypeptides that govern the communication between cells within the multicellular organism. The technology is presently developing in two directions: (1) initial studies were done with small peptides, then progressed to medium-size polypeptides, and are now more and more utilized for proteins, and (2) less and less material is becoming necessary for structural characterization by amino acid analysis and sequencing, i.e., whereas initially several nanomoles of purified material were necessary, picomolar amounts are now required for partial or complete structural characterization. Over the years, reverse-phase liquid chromatography (RPLC) has evolved as the dominant tool for the isolation and characterization of polypeptides. Recently, however, alternative modes of chromatography, such as high-performance ion exchange, gel filtration, and chromatofocusing techniques, have been further developed, thus becoming integral parts of isolation programs.

Keywords

Organic Modifier Teflon Tubing Sampling Valve Slider Valve Organic Solvent Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© The Humana Press Inc. 1986

Authors and Affiliations

  • Petro E. Petrides
    • 1
  1. 1.Munich University School of MedicineGrossharden, MunichWest Germany

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