Strategies for Protein Purification

R.K. Scopes1

1 La Trobe University, Bundoora
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 1.2
DOI:  10.1002/0471140864.ps0102s00
Online Posting Date:  May, 2001
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

This discussion unit presents information on classification of proteins according to shape, oligomeric structure, and function. In addition, strategies for purifying different types of proteins are discussed based on their location and associations within the cell. Specifically, critical issues are detailed for extracellular, cytoplasmic, and membraneā€associated proteins, as well as for insoluble proteins. Finally, strategies for the purification of both soluble and insoluble (inclusion bodies) recombinantly expressed proteins are provided.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Classification of Proteins
  • Strategies for Protein Purification
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Oka, M., Yang, Y.S., Nagata, S., Esaki, N., Tanaka, M., and Soda, K. 1989. Overproduction of thermostable leucine dehydrogenase of Bacillus stearothermophilus and its one‐step purification from recombinant cells of. Escherichia coli. Biotechnol. Appl. Biochem. 11:307‐316.
   Sassenfeld, H.M. 1990. Engineering proteins for purification. Trends Biotechnol. 8:88‐93.
Key References
   Deutscher, M.P., (ed.) 1990. Guide to protein purification. Methods Enzymol 182:1‐894.
  Extensive collection of purification methods with some general protocols and examples.
   Janson, J.‐C. and Ryden, L.G. 1989. Protein Purification: Principles, High Resolution Methods, and Applications. VCH Publishers, New York.
  A useful collection of methods and examples.
  Kennedy, J.F. and Cabral, J.M., (eds.) 1993. Recovery Processes for Biological Materials. John Wiley & Sons, New York.
  A useful introduction to the problems of large‐scale methods.
   Kenny, A. and Fowell, S., (eds.) 1992. Practical protein chromatography. Methods Mol. Biol. 11:1‐327.
  Extensive descriptions of affinity chromatographic techniques with protocols and recipes.
   Scopes, R.K. 1993. Protein Purification, Principles and Practice, 3rd ed. Springer‐Verlag, New York and Heidelberg.
  General principles of all the main techniques used in purifying proteins. A useful laboratory handbook; does not include recipes or procedures for specific proteins.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library