Absolute quantification of the model biomarker prostate-specific antigen in serum by LC-Ms/MS using protein cleavage and isotope dilution mass spectrometry

Journal of Proteome Research
David R BarnidgeDavid C Muddiman

Abstract

Protein cleavage-isotope dilution mass spectrometry (PC-IDMS) can be used to quantify proteins, with an isotope-labeled analogue of the peptide fragment used as an internal standard. Here, we investigate use of a standard LC-MS/MS platform for quantifying a model biomarker directly from serum by this technique. We synthesized a peptide (IVGGWECEK) identical to the N-terminal tryptic fragment of PSA but with each glycine containing two 13C atoms and one 15N atom. PSA-free human serum was denatured with urea followed by the introduction of PSA standard and the stable isotope labeled internal standard peptide. The sample was then proteolyzed with trypsin and subjected to quantification using LC-MS/ MS on a triple quadrupole mass spectrometer. A linear least squares calibration curve made from five different concentrations of PSA added to serum and digested (each made in triplicate and randomly injected three times) had a mean slope of 0.973 (SE = 0.023), intercept of -0.003 (SE = 0.022), and R2 of 0.971. Recovery of calibrators ranged from 70 to 85% with a mean run-to-run CV of 13% and a mean within-run CV of 5.7%. PC-IDMS is a promising technique for quantifying proteins covering a broad range of applications from standardizing i...Continue Reading

References

Mar 28, 2002·Clinical Chemistry and Laboratory Medicine : CCLM·Jan-Olof JeppssonUNKNOWN International Federation of Clinical Chemistry and Laboratory Medicine (IFCC)
Jan 25, 2003·Molecular & Cellular Proteomics : MCP·Joshua N AdkinsJoel G Pounds
Apr 30, 2003·Journal of Mass Spectrometry : JMS·R KostiainenS Auriola
Oct 6, 1989·Science·B F Chmelka, A Pines

❮ Previous
Next ❯

Citations

Jun 28, 2008·Analytical and Bioanalytical Chemistry·H SchlüterM Trusch
Mar 24, 2009·Analytical and Bioanalytical Chemistry·Luciano CallipoAldo Laganà
Jul 20, 2010·Analytical and Bioanalytical Chemistry·Adam M HawkridgeDavid C Muddiman
Mar 30, 2011·Analytical and Bioanalytical Chemistry·Fabien XuerebJean-Marie Schmitter
Jan 18, 2013·Analytical and Bioanalytical Chemistry·Quentin EnjalbertJérôme Lemoine
Sep 8, 2009·Journal of the American Society for Mass Spectrometry·D Keith WilliamsDavid C Muddiman
Sep 23, 2008·Journal of Proteome Research·Kelli G KlineChristine C Wu
Dec 25, 2008·Journal of Proteome Research·Sheng PanTeresa A Brentnall
Jun 21, 2011·Nature Biotechnology·Jeffrey R WhiteakerAmanda G Paulovich
Aug 19, 2011·Molecular & Cellular Proteomics : MCP·Yun XiangJohn M Koomen
Jan 20, 2011·Molecular & Cellular Proteomics : MCP·Jeffrey R WhiteakerAmanda G Paulovich
Mar 24, 2012·Molecular & Cellular Proteomics : MCP·Yaojun LiNing Li
Jun 27, 2009·Molecular & Cellular Proteomics : MCP·Jamie ShermanMark P Molloy
May 5, 2009·Molecular & Cellular Proteomics : MCP·Michael A KuzykChristoph H Borchers
Mar 13, 2009·Briefings in Functional Genomics & Proteomics·Anastasia K Yocum, Arul M Chinnaiyan
Apr 29, 2009·Briefings in Functional Genomics & Proteomics·Kelli G KlineChristine C Wu
Apr 24, 2012·Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association·Vesna D GarovicDavid R Barnidge
Jul 19, 2008·Annual Review of Analytical Chemistry·Bradley L AckermannAjai K Chaudhary
May 14, 2010·Annual Review of Biophysics·Kelli G Kline, Michael R Sussman
Nov 28, 2007·Annual Review of Pathology·Raghothama Chaerkady, Akhilesh Pandey
Apr 8, 2009·Proceedings of the American Thoracic Society·Sebahat OcakPierre P Massion
Mar 31, 2010·Bioanalysis·Eugene Ciccimaro, Ian A Blair
May 10, 2011·Annual Review of Biochemistry·Jürgen Cox, Matthias Mann
Aug 23, 2011·Journal of Proteomics·Jinong LiZivana Tezak
Mar 28, 2009·Clinical Chemistry·Jesse C SeegmillerRajiv Kumar
Feb 27, 2013·Analytical Chemistry·Yunli HuYehia Mechref
Dec 22, 2006·Expert Review of Proteomics·Linfeng Wu, David K Han

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.