On-column trypsin digestion coupled with LC-MS/MS for quantification of apolipoproteins

Toth, Christopher; Kuklenyik, Zsuzsanna; Jones, Jeffrey I.; Parks, Bryan A.; Gardner, Michael S.; Schieltz, David; Rees, Jon C.; Andrews, Michael; McWilliams, Lisa G.; Pirkle, James L.; Barr, John R.

doi:10.1016/j.jprot.2016.09.011

Cited by 48 publications

(34 citation statements)

References 42 publications

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“…Targeted proteomics assays using LC‐MS/MS for a specific protein quantitation are most promising approaches for an implementation of a comprehensive apo analysis into routine diagnostics, but most workflows suffer from laborious and time‐consuming manual sample preparation . For the quantitation of apos, several LC‐MS/MS assays have been developed in recent years . Although some of those forwent clean‐up steps used for a removal of interfering salts and detergents prior to mass spectrometric peptide analysis, most applications utilize off‐line SPE or stable isotope standards and capture by anti‐peptide antibodies (SISCAPA) for desalting and enrichment of samples.…”

Section: Discussionmentioning

confidence: 99%

“…Signature peptides of apos C‐II, D, and H were selected from literature to extend a previously published off‐line SPE‐LC‐MS/MS method . Further, potential signature peptides for apoM and apoJ were identified using UniProt sequence data in combination with accepted selection criteria like peptide lengths of 7‐20 amino acids and omission of known post‐translational modifications, polymorphisms and missed cleavage sites .…”

Section: Methodsmentioning

confidence: 99%

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Targeted On‐line SPE‐LC‐MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood

et al. 2018

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Laborious sample pretreatment of biological samples represents the most limiting factor for the translation of targeted proteomics assays from research to clinical routine. An optimized method for the simultaneous quantitation of 12 major apolipoproteins (apos) combining on-line SPE and fast LC-MS/MS analysis in 6.5 min total run time was developed, reducing the manual sample pretreatment time of 3 μL serum or plasma by 60%. Within-run and between-day imprecisions below 10 and 15% (n = 10) and high recovery rates (94-131%) were obtained applying the high-throughput setup. High-quality porcine trypsin was used, which outperformed cost-effective bovine trypsin regarding digestion efficiency. Comparisons with immunoassays and another LC-MS/MS assay demonstrated good correlation (Pearson's R: 0.81-0.98). Further, requirements on sample quality concerning sampling, processing, and long-term storage up to 1 year were investigated revealing significant influences of the applied sampling material and coagulant on quantitation results. Apo profiles of 1339 subjects of the LIFE-Adult-Study were associated with lifestyle and physiological parameters as well as establish parameters of lipid metabolism (e.g., triglycerides, cholesterol). Besides gender effects, most significant impact was seen regarding lipid-lowering medication. In conclusion, this novel highly standardized, high-throughput targeted proteomics assay utilizes a fast, simultaneous analysis of 12 apos from least sample amounts.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Targeted On‐line SPE‐LC‐MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood

et al. 2018

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“…Apolipoprotein quantification by liquid chromatography ID/MS (LC-ID/MS) was first reported in the late 1990s by Barr et al for apoA-I [58] and was further applied to other apolipoproteins (apoB, C and E) in the following years [59][60][61].…”

Section: Apolipoprotein Profiling By Liquid Chromatography Isotopic-dmentioning

confidence: 99%

“…After digestion, apolipoprotein-specific tryptic peptides were identified for each major class of apolipoproteins, and some of them were selected for quantification by ID/MS [60][61][62]. ID/MS quantification uses synthetic labeled entities with 13 C, 15 N or deuterium as internal standards (IS) to spike the samples.…”

Section: Apolipoprotein Profiling By Liquid Chromatography Isotopic-dmentioning

confidence: 99%

“…These materials were endorsed by the World Health Organization (WHO) and widely used to recalibrate routine apolipoprotein immunoassays but were never intended for standardization purposes. It appears nevertheless that most ID-LC/MS methods for apolipoprotein quantification use these WHO RMs as external calibrators [60][61][62].…”

Section: Apolipoprotein Profiling By Liquid Chromatography Isotopic-dmentioning

confidence: 99%

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Advanced lipoprotein testing for cardiovascular diseases risk assessment: a review of the novel approaches in lipoprotein profiling

Clouet-Foraison

Gaie-Levrel

Gillery

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Abstract:With the increasing prevalence of cardiovascular diseases (CVD) worldwide, finding reliable and clinically relevant biomarkers to predict acute cardiovascular events has been a major aim of the scientific and medical community. Improvements of the understanding of the pathophysiological pathways of the disease highlighted the major role of lipoprotein particles, and these past decades have seen the emergence of a number of new methodologies to separate, measure and quantitate lipoproteins. Those methods, also known as advanced lipoprotein testing methods (ALT), have gained acceptance in the field of CVD risk assessment and have proven their clinical relevance. In the context of worldwide standardization and harmonization of biological assays, efforts have been initiated toward standardization of ALT methods. However, the complexity of lipoprotein particles and the multiple approaches and methodologies reported to quantify them have rendered these initiatives a critical issue. In this context and to better understand these challenges, this review presents a summary of the major methods available for ALT with the aim to point out the major differences in terms of procedures and quantities actually measured and to discuss the resulting comparability issues.

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The small HDL particle hypothesis of Alzheimer's disease

Martinez

Weissberger

Kuklenyik

et al. 2022

Alzheimer's & Dementia

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We propose the hypothesis that small high‐density lipoprotein (HDL) particles reduce the risk of Alzheimer's disease (AD) by virtue of their capacity to exchange lipids, affecting neuronal membrane composition and vascular and synaptic functions. Concentrations of small HDLs in cerebrospinal fluid (CSF) and plasma were measured in 180 individuals ≥60 years of age using ion mobility methodology. Small HDL concentrations in CSF were positively associated with performance in three domains of cognitive function independent of apolipoprotein E (APOE) ε4 status, age, sex, and years of education. Moreover, there was a significant correlation between levels of small HDLs in CSF and plasma. Further studies will be aimed at determining whether specific components of small HDL exchange across the blood, brain, and CSF barriers, and developing approaches to exploit small HDLs for therapeutic purposes.

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On-column trypsin digestion coupled with LC-MS/MS for quantification of apolipoproteins

Cited by 48 publications

References 42 publications

Targeted On‐line SPE‐LC‐MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood

Targeted On‐line SPE‐LC‐MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood

Advanced lipoprotein testing for cardiovascular diseases risk assessment: a review of the novel approaches in lipoprotein profiling

The small HDL particle hypothesis of Alzheimer's disease

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