Protein purification
- 1 January 1989
- book chapter
- Published by Springer Nature
- Vol. 40, 19-71
- https://doi.org/10.1007/bfb0009827
Abstract
This monograph summarizes recent developments in the purification and analysis of natural and recombinant proteins. The basic strategies employed in protein purification are reviewed with regards to the characteristics of the protein of interest that may aid its isolation, choice of the starting material, and use of denaturants. Preparation of cell-free extracts followed by bulk precipitation and/or phase partition constitute the initial steps of many purification schemes. Chromatographic methods (size exclusion, ion exchange, hydroxylapatite, reversed phase, hydrophobic interaction and affinity based) utilizing either traditional, low pressure or high-performance liquid chromatography instrumentation are discussed. Electrophoretic techniques used to analyze the homogeneity of the protein product include SDS-PAGE, isotachophoresis, IEF and two dimensional gel electrophoresis.Keywords
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