An integrated top-down and bottom-up strategy for broadly characterizing protein isoforms and modifications

February 10, 2009

Journal Article

An integrated top-down and bottom-up strategy for broadly characterizing protein isoforms and modifications

Abstract

We present an integrated top-down and bottom-up approach facilitated by concurrent liquid chromatography-mass spectrometry (LC-MS) analysis and fraction collection for comprehensive high-throughput intact protein profiling. The approach employs high resolution reversed phase (RP) LC separations coupled on-line with a 12T Fourier transform ion cyclotron resonance (FTICR) spectrometer to profile and tentatively identify modified proteins, using detected intact protein masses in conjunction with bare protein identifications from the bottom-up analysis of the same fraction. Selected identifications are incorporated into a target ion list for subsequent offline gas phase fragmentation that uses only an aliquot of the original fraction used for bottom-up analysis.

Revised: April 24, 2009 | Published: February 10, 2009

Citation

Wu S., N.M. Lourette, N. Tolic, R. Zhao, R. Robinson, A.V. Tolmachev, and R.D. Smith, et al. 2009. An integrated top-down and bottom-up strategy for broadly characterizing protein isoforms and modifications. Journal of Proteome Research 8, no. 3:1347-1357. PNNL-SA-58633.