Development of novel tags for enhanced sample multiplexing and application of cPILOT to understand aging mechanisms
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1
Vanderbilt University, United States
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2
Texas Southern University, United States
Large-scale quantitative proteomics can provide a near-global view of cellular protein abundance. To fully leverage mass spectrometry for the elucidation of disease processes high-throughput proteomics is necessary to monitor dynamic changes across samples from varying conditions. One way to do this, is with the use of isobaric tagging strategies, which allow as many as 12 samples to be multiplexed in a single analysis. Through an approach which has dual tagging nature, combines isotopic labeling with isobaric tagging (cPILOT), the capacity of samples doubles. The goal of this study is to improve sample throughput of our combined precursor isotopic labeling and isobaric tagging (cPILOT) approach. The major challenge to making this approach work involve developing amine tagging reagents specific to either N-terminal or lysine amine residues. To overcome this challenge, we are building on previous studies in our laboratory, and developing novel strategies. This presentation will cover preliminary findings to date. Moreover, we have successfully applied cPILOT approach to study metabolic pathways involved in the aging process. cPILOT has enabled us to analyze 14 samples in a single shot for the identification of 2997 and quantification of 1814 protein groups in rabbit liver. The implications of this study will be presented.
Keywords:
multiplexing,
cPILOT,
Novel Tags Development,
Quantitative proteome analysis,
Aging,
Peptide-based tags,
amine tagging reagents
Conference:
National Organization for the Professional Advancement of Black Chemists and Chemical Engineers (NOBCChE) 45th Annual Conference , Orlando, Florida, United States, 17 Sep - 20 Sep, 2018.
Presentation Type:
Oral Presentation
Topic:
Analytical Chemsitry
Citation:
Amin
B,
Daniels
J and
Robinson
RA
(2019). Development of novel tags for enhanced sample multiplexing and application of cPILOT to understand aging mechanisms.
Front. Chem.
Conference Abstract:
National Organization for the Professional Advancement of Black Chemists and Chemical Engineers (NOBCChE) 45th Annual Conference .
doi: 10.3389/conf.fchem.2018.01.00037
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Received:
09 Oct 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Prof. Renã A Robinson, Vanderbilt University, Nashville, United States, rena.as.robinson@vanderbilt.edu