Overview of Conventional Chromatography

Alan Williams1

1 Pharmacia Biotech, Piscataway
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 8.1
DOI:  10.1002/0471140864.ps0801s00
Online Posting Date:  May, 2001
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Abstract

The key to success in purification of any protein is developing the right separation strategy. This involves selecting and correctly applying a combination of separation techniques, chosen on the basis of information about the target protein and contaminants. There is no single separation technique and no single purification scheme that will allow successful purification of all types of proteins. However, there is a single approach to developing a successful purification strategy. This overview discusses the basic steps that must be taken, including selection of source material, and progression from initial “capture” stage of separation to the final polishing stage. Factors that can affect chromatographic resolution are also covered.

     
 
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Table of Contents

  • Yield and Purity of Target Protein
  • Steps in a Purification Strategy
  • Parameters Affecting Chromatographic Resolution
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Figures

Videos

Literature Cited

Literature Cited
   Corran, P.H. 1989. Reversed‐phase chromatography of proteins. In HPLC of Macromolecules: A Practical Approach (R.W.A. Oliver, ed.) pp. 127‐156. IRL Press, Oxford.
   Giddings, J.C. and Keller, R.A. (eds.) 1965. Dynamics of Chromatography, Part 1: Principles and Theory. Marcel Dekker, New York.
   Janson, J.‐C. and Ryden, L. (eds.) 1989. Protein Purification: Principles, High Resolution Methods and Applications. VCH Publishers, New York.
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