• Shigella dysenteriae
• Whole Cell Lysate
• Staphylococcus aureus
• Periplasm, Cell Envelope & Cell Wall
• Streptococcus pneumoniae
• Whole Cell Lysate
• Yersinia pestis
• Periplasm, Cell Envelope & Cell Wall
• Membrane
• Poster Presentations

High quality protein microarray using in situ protein purification

¹, Carissa Grose¹, Rembert Pieper¹, Gagan A. Pandya¹, Robert D Fleischmann¹ and Scott N Peterson¹

¹Pathogen Functional Genomics Resource Center, J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA

Abstract

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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