Hydrophilic interaction liquid chromatography as second dimension in multidimensional chromatography with an anionic trapping strategy: application to prostate-specific antigen quantification.

Romain Simon; Sébastien Passeron; Jérôme Lemoine; Arnaud Salvador

doi:10.1016/j.chroma.2014.05.063

Hydrophilic interaction liquid chromatography as second dimension in multidimensional chromatography with an anionic trapping strategy: application to prostate-specific antigen quantification.

Romain Simon ¹ Sébastien Passeron ² Jérôme Lemoine ¹ Arnaud Salvador ^{1, *}

* Corresponding author

1 ANABIO-MS - Analyse biomoléculaire par spectrométrie de masse - Biological Analysis by Mass Spectrometry

ISA - Institut des Sciences Analytiques

2 ISA - Institut des Sciences Analytiques

Abstract : Liquid chromatography (LC) coupled with tandem mass spectrometry (MS-MS) in selected reaction monitoring mode (SRM) has become a widely used technique for the quantification of protein biomarkers in plasma and has already proven to give similar results compared to the conventional immunoassays. To improve the lack of insufficient sensitivity for quantification of low abundance protein, we propose a new two dimensional liquid chromatography (2D-LC-SRM) method for the quantitation of prostate specific antigen (PSA) in human plasma. The method centers on anion exchange cartridge between reversed-phase chromatography and hydrophilic interaction liquid chromatography (HILIC) in an on-line arrangement. The use of the anionic cartridge allows an easier online transfer of the analytes between both dimensions. Moreover, it provides an additional selectivity since the more basic peptides are not retained on this support. This setup has been applied to the quantification of prostate specific antigen (PSA) protein in plasma on a previous generation of mass spectrometer, which enabled a limit of quantification (LOQ) of 1ng/mL without any upfront immuno-depletion or intense off-line fractionation before the SRM analysis. The obtained LOQ is compatible with the required sensitivity for the clinically relevant plasma-based PSA tests.

Keywords : 2D-LC HILIC Mass spectrometry Prostate specific antigen Protein Quantification TARGETED MASS-SPECTROMETRY LOW-ABUNDANCE PROTEINS IMMUNOAFFINITY DEPLETION BIOMARKER VALIDATION HUMAN PLASMA PHOTO-SRM MRM QUANTITATION PEPTIDES PHASE

Document type :

Journal articles

Domain :

Chemical Sciences / Analytical chemistry

Life Sciences [q-bio] / Biochemistry, Molecular Biology / Biomolecules [q-bio.BM]

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https://hal.archives-ouvertes.fr/hal-01071821
Contributor : Agnès Bussy <>
Submitted on : Monday, October 6, 2014 - 5:03:41 PM
Last modification on : Thursday, February 8, 2018 - 11:10:27 AM

Hydrophilic interaction liquid chromatography as second dimension in multidimensional chromatography with an anionic trapping strategy: application to prostate-specific antigen quantification.

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Hydrophilic interaction liquid chromatography as second dimension in multidimensional chromatography with an anionic trapping strategy: application to prostate-specific antigen quantification.

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