Quantification of Proteins and Metabolites by Mass Spectrometry without Isotopic Labeling or Spiked Standards

Weixun Wang,† Haihong Zhou,† Hua Lin, Sushmita Roy, Thomas A. Shaler, Lander R. Hill, Scott Norton, Praveen Kumar, Markus Anderle, and Christopher H. Becker*
SurroMed, Inc., 2375 Garcia Avenue, Mountain View, California 94043
Anal. Chem., 2003, 75 (18), pp 4818–4826
DOI: 10.1021/ac026468x
Publication Date (Web): August 8, 2003
Copyright © 2003 American Chemical Society
†

 These two authors contributed equally to this work.

,
*

 Corresponding author. Phone:  (650) 230-1845. Fax:  (650) 230-1960. E-mail:  cbecker@surromed.com.

Abstract

A new method is presented for quantifying proteomic and metabolomic profile data by liquid chromatography−mass spectrometry (LC−MS) with electrospray ionization. This biotechnology provides differential expression measurements and enables the discovery of biological markers (biomarkers). Work presented here uses human serum but is applicable to any fluid or tissue. The approach relies on linearity of signal versus molecular concentration and reproducibility of sample processing. There is no use of isotopic labeling or chemically similar standard materials. Linear standard curves are reported for a variety of compounds introduced into human serum. As a measure of analytical reproducibility for proteome and metabolome sampling, median coefficients of variation of 25.7 and 23.8%, respectively, were determined for 3400 molecular ions (not counting their numerous isotopes) from 25 independently processed human serum samples, corresponding to a total of 85 000 individual molecular ion measurements.

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History

  • Published In Issue September 15, 2003
  • Received for review December 26, 2002. Accepted July 1, 2003.

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