High resolution time-of-flight mass analysis of the entire range of intact singly-charged proteins

Jeonghoon Lee; Huijuan Chen; Tiancheng Liu; Clifford E Berkman; Peter T A Reilly

doi:10.1021/ac202001z

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High resolution time-of-flight mass analysis of the entire range of intact singly-charged proteins

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High resolution time-of-flight mass analysis of the entire range of intact singly-charged proteins

Jeonghoon Lee, Huijuan Chen, Tiancheng Liu, Clifford E Berkman and Peter T A Reilly

Analytical chemistry (Washington), Vol.83(24), pp.9406-9412

12/15/2011

DOI: https://doi.org/10.1021/ac202001z

Handle:

https://hdl.handle.net/2376/105148

PMCID: PMC3237766

PMID: 22047146

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Abstract

Electrodes

Streptavidin - chemistry

Carbocyanines - chemistry

Ions - chemistry

Biotinylation

Molecular Weight

Proteins - analysis

Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Atmospheric Pressure

Peptides - analysis

The proof of principle for high-resolution analysis of intact singly charged proteins of any size is presented. Singly charged protein ions were produced by electrospray ionization followed by surface-induced charge reduction at atmospheric pressure. The inlet and trapping system "stops" the forward momentum of the protein ions over a very broad range to be captured by the digitally produced electric fields of a large radius linear ion trap whereupon they are moved into a smaller radius linear ion trap and collected and concentrated in front of its exit end-cap electrode using digital waveform manipulation. The protein ions are then ejected on demand from the end of the small radius linear quadrupole in a tightly collimated ion beam with an instrumentally defined kinetic energy into the acceleration region of an orthogonal acceleration reflectron time-of-flight mass analyzer where their flight times were measured and detected with a Photonis BiPolar TOF detector. We present results that clearly prove that massive singly charged ions can yield high-resolution mass spectra with very low chemical noise and without loss of sensitivity with increasing mass across the entire spectrum. Analysis of noncovalently bound protein complexes was demonstrated with streptavidin-Cy5 bound with a biotinylated peptide mimic. Our results suggest proteins across the entire range can be directly quantified using our mass analysis technique. We present evidence that solvent molecules noncovalently adduct onto the proteins while yielding consistent flight time distributions. Finally, we provide a look into future that will result from the ability to rapidly measure and quantify protein distributions.

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Title: High resolution time-of-flight mass analysis of the entire range of intact singly-charged proteins
Creators: Jeonghoon Lee - Washington State University, Department of Chemistry, Pullman, Washington 99164, United States
Huijuan Chen
Tiancheng Liu
Clifford E Berkman
Peter T A Reilly
Publication Details: Analytical chemistry (Washington), Vol.83(24), pp.9406-9412
Academic Unit: Chemistry, Department of
Publisher: United States
Grant note: R01 GM088501 / NIGMS NIH HHS R01 CA140617 / NCI NIH HHS R01 GM088501-01 / NIGMS NIH HHS 5R01CA140617-02 / NCI NIH HHS
Identifiers: 99900546758401842
Language: English
Resource Type: Journal article