LC-MS-Based Quantification of Intact Proteins: Perspective for Clinical and Bioanalytical Applications
Mass spectrometry as a routine tool in the clinic is becoming more and more of a reality every day, presenting numerous opportunities in the field of proteomics. This review provides a comprehensive overview of existing LC-MS bottom-up proteomic techniques as well as the potential for future approaches using intact protein quantification. It includes a comparison between the two approaches, the limitations of both, and some of the hurdles that will need to be overcome for successful detection.
Abstract
Irene van den Broek and William D. van Dongen
Bioanalysis, Vol. 7, No. 15 , Pages 1943-1958 (doi: 10.4155/bio.15.113)
Bioanalytical LC–MS for protein quantification is traditionally based on enzymatic digestion of the target protein followed by absolute quantification of a specific signature peptide relative to a stable-isotope labeled analog. The enzymatic digestion, nonetheless, limits rapid method development, sample throughput and turnaround time, and, moreover, makes that essential information regarding the biological function of the intact protein is lost. The recent advancements in high-resolution MS instrumentation and improved sample preparation techniques dedicated to protein clean-up raise the question to what extent LC–MS can be applied for quantitative bioanalysis of intact proteins. This review provides an overview of current and potential applications of LC–MS for intact protein quantification as well as the main limitations and challenges for broad application.
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