PO0006 Automated Immunoaffinity Enrichment for Phosphotyrosine Peptides Using Protein A andStreptavidin IMCStips
来源:IMCS
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作者:Sunil Hwang*1,Todd Mullis2, Michael Walla2, Matthew Manter1, Jingyun Lee3, Christina M Furdui3, L. Andrew Lee1
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发布时间: 2059天前
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2416 次浏览
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•Established automated Protein A and Streptavidin immunoaffinity enrichment protocol for high-
throughput sample preparation
Identified over 1000 phosphotyrosine peptides using anti-phosphotyrosine antibodies and Protein A /Streptavidin IMCStips
•Achieved over 94% specificity of phosphotyrosine peptide enrichment using Hamilton® Microlab STAR workstation and IMCStips
throughput sample preparation
Identified over 1000 phosphotyrosine peptides using anti-phosphotyrosine antibodies and Protein A /Streptavidin IMCStips
•Achieved over 94% specificity of phosphotyrosine peptide enrichment using Hamilton® Microlab STAR workstation and IMCStips
Signal transduction of protein phosphorylation is a key mechanism for survival, maintenance, and deathof eukaryotic cells (1-4). Te global enrichment of phosphoserine and phosphothreonine peptides hasbeen successfully optimized using immobilized metal affinity chromatography (IMAC) and titaniumdioxide (TiO2) using IMCStips with an automated liquid handling system. However, enrichment ofphosphotyrosine (p-Tyr) peptides are less efficient with these methods due to the low stoichiometry of p-Tyr peptides to global phosphorylation event (5-11). Here, we introduce anti-phosphotyrosineantibody enrichment using a dispersive pipette extraction technology in IMCStips. Te method uses anautomatic liquid handling system to enrich p-Tyr peptides for high-throughput and reproducible samplepreparation.
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