Proteome Based Improvements for Immobilized Metal Affinity Chromatography in an Esherichia Coli Expression System

Proteome Based Improvements for Immobilized Metal Affinity Chromatography in an Esherichia Coli Expression System

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E. coli expression systems are important tools for structural and functional analysis in research and industry. Adding an affinity tag is attractive for a target protein with unknown purification characteristics. Indeed, more than 60% of proteins for structural studies are produced with a poly-histidine tag (his-tag). Naturally occurring histidine and cysteine rich regions in host cell proteins result in unwanted protein binding during an Immobilized Metal Affininty Chromatography (IMAC) capture step. In this dissertation host proteins which have affinity to IMAC columns were identified and characterized. Data will be helpful in understanding and controlling contaminants that elute with his-tagged proteins.1 Number of genes present in Total contaminant pools, essential and non- essential of Co(II), Ni(II), Zn(II) IMAC for pH binding condition 99 Figure 6.2 Number of genes ... Scope of work Immobilized metal affinity chromatography ( IMAC) is xiii.


Title:Proteome Based Improvements for Immobilized Metal Affinity Chromatography in an Esherichia Coli Expression System
Author: Rajaramesh Varakala
Publisher:ProQuest - 2008
ISBN-13:

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