Final ProgramCPSA 2011
Science and Technology Coming Together to Make a Difference
October 3 - 6, 2011
Bucks County Sheraton Hotel
Langhorne, PA
Poster Abstract #14
Evaluation of 150 µm ID Packed Tip Columns for Quantitative Peptide Analysis by LC-MS/MS
New Objective, Inc., Woburn, MA USA
A predominant workflow for qualitative proteomics has been "GeLC-MS," a combination of 1- (or 2-D) gel electrophoresis with reverse-phase nanoflow liquid chromatography mass spectrometry (nLC-MS/MS). The limited protein quantity isolated from a single gel band coupled with column loading capacity maximums necessitate the use of 75 µm ID packed columns for optimal sensitivity. However, limitations on sample injection volume, gradient and flow characteristics, and excessive delay volume hinder throughput. Novel methods for fractionating complex biological samples with higher loading capacities and more efficient recovery, such as novel solution phase tube-gel fractionation and others, demand a column format which maximizes on the extended dynamic range of these emerging techniques. Packed tip columns with a larger ID (150 µm to 200 µm) facilitate higher sample loading capacity and enable higher flow rates for improved cycle time while maintaining the optimal sensitivity realized in the nanobore packed tip columm format. Using peptide standards, single protein digests and whole yeast digests improvements in cycle time and sample loading capacity using 150 µm ID packed tip columns are demonstrated.