High quality protein microarray using in situ protein purification |
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Authors: | Keehwan Kwon Carissa Grose Rembert Pieper Gagan A Pandya Robert D Fleischmann and Scott N Peterson |
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Institution: | (1) Pathogen Functional Genomics Resource Center, J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, USA |
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Abstract: | Background In the postgenomic era, high throughput protein expression and protein microarray technologies have progressed markedly permitting
screening of therapeutic reagents and discovery of novel protein functions. Hexa-histidine is one of the most commonly used
fusion tags for protein expression due to its small size and convenient purification via immobilized metal ion affinity chromatography
(IMAC). This purification process has been adapted to the protein microarray format, but the quality of in situ His-tagged protein purification on slides has not been systematically evaluated. We established methods to determine the
level of purification of such proteins on metal chelate-modified slide surfaces. Optimized in situ purification of His-tagged recombinant proteins has the potential to become the new gold standard for cost-effective generation
of high-quality and high-density protein microarrays. |
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