Multistep solid phase mass tagging coupled with 2-D LC/MS strategy for quantitative proteomics

ANYL 509

Yang Shi, yang.shi@yale.edu, Rong Xiang, rong.xiang@yale.edu, Csaba Horváth, csaba_horvath@yale.edu, and James A. Wilkins, james.wilkins@yale.edu. Department of Chemical Engineering, Yale University, 9 Hillhouse Avenue, New Haven, CT 06520-8286
Identification of large numbers of proteins from complex biological samples is a continuing challenge in the area of quantitative proteomics. We introduce here a simple and reliable multistep mass tagging technique using our recently developed solid phase mass tagging reagents. When coupled with 2D LC/nano ESI-MS, this method allows enhanced protein identification when tested on samples from prokaryotic and eukaryotic sources. The proteome of Escherichia coli D21 grown to either mid-exponential or stationary phase, and the membrane proteome from established breast cancer cell lines BT474 and MCF7 were used as model systems in these experiments. In both experiments, the numbers of total identified proteins are at least twice the numbers identified from a single tagging cycle. The sample complexity can be effectively reduced with corresponding increases in protein identification using the multistep method. The strategy described here represents a potentially powerful technique for large-scale qualitative and quantitative proteome research.
 

General Papers
1:30 PM-4:50 PM, Thursday, 1 September 2005 Washington DC Convention Center -- 153, Oral

Sci-Mix
8:00 PM-10:00 PM, Monday, 29 August 2005 Washington DC Convention Center -- Hall A, Sci-Mix

Division of Analytical Chemistry

The 230th ACS National Meeting, in Washington, DC, Aug 28-Sept 1, 2005