Affinity labeling coupled with matrix assistant laser desorption tandem time of flight mass spectrometry for quantitative proteomics research

Abstract

A relative quantitative method for differential proteomics by cleavable isotope-coded affinity tag (cICAT) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-MS) was established. The accuracy and reproducibility of the method were evaluated by bovine serum albumin (BSA) digest as having a relative standard deviation of less than 30% and good reproducibility. The dynamic range was also evaluated by analyzing two mixtures of several standard proteins with different concentration. The experimental results showed that in the dynamic range of 1:30, the quantitation error of the method was less than 30%. Although the quantitation error becomes very large when used beyond this range, it does not affect the derivation of information on the differential proteins. All the work provides an alternative method for differential proteomics analysis in biological samples from different origins.

Keywords

isotope coded affinity tag, matrix assistant laser desorption ionization tandem time of flight mass spectrometry, capillary reversed-phase liquid chromatography