High precision quantification of human plasma proteins using the automated SISCAPA Immuno-MS workflow

Razavi, Morteza; Anderson, N. Leigh; Pope, Matthew E.; Yip, Richard; Pearson, Terry W.

doi:10.1016/j.nbt.2015.12.008

Cited by 61 publications

(68 citation statements)

References 22 publications

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“…The stage at which such reagents are used is governed by the nature of capture. In general, anti-protein antibodies are incorporated at the beginning of an analytical workflow for protein enrichment [39,49,50], while anti-peptide antibodies are employed post-digestion for tryptic peptide enrichment [51][52][53] (see Figure 2). In an example of the former, anti-protein antibodies were covalently bound to a monolithic column (as opposed to magnetic beads) [50] for protein enrichment and detection of the target proteins or peptides in a top-down [54] or bottom-up [55] manner, respectively.…”

Section: Status and Challenges Of Current Methodsmentioning

confidence: 99%

“…Over the past few years, this approach has been applied to precisely quantify panels of disease-related proteins in human plasma [51,52,[61][62][63][64][65]. In a recent study, 40 novel immuno-MRM assays (for 27 proteins) were developed and validated in accordance with the characterization guidelines for Tier 2 assays [66].…”

Section: Status and Challenges Of Current Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Clinical translation of MS-based, quantitative plasma proteomics: status, challenges, requirements, and potential

Percy¹,

Byrns

Pennington

et al. 2016

Expert Review of Proteomics

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This article reviews the status, challenges, requirements, and potential of translating current, MS-based methods to the clinical laboratory. The described methods are discussed and contrasted within a fit-for-purpose approach, while different resources for quality control, quantitative analysis, and data interpretation are additionally provided. Expert commentary: Although great strides have been made over the past five years in developing reliable quantitative assays for plasma protein biomarkers, it is crucial for investigators to have an understanding of the clinical validation process, a major roadblock in translational research. Continued progress in method design and validation of protein assays is necessary to ultimately achieve widespread adoption and regulatory approval.

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Section: Status and Challenges Of Current Methodsmentioning

confidence: 99%

Section: Status and Challenges Of Current Methodsmentioning

confidence: 99%

Clinical translation of MS-based, quantitative plasma proteomics: status, challenges, requirements, and potential

Percy¹,

Byrns

Pennington

et al. 2016

Expert Review of Proteomics

View full text Add to dashboard Cite

show abstract

“…Enrichment with antibodies can take place at the protein level, with applications in elucidating transcription regulatory networks, cell signaling pathways, and biomarker discovery, or postdigestion at the peptide level for peptide enrichment before LC‐MS/MS . An example of such technique is Stable Isotope Standards and Capture by Anti‐Peptide Antibodies (SISCAPA), also referred to as immuno‐selected reaction monitoring technology . This approach allows reduction of matrix complexity within a whole‐cell digest by enriching only the desired peptides, followed by MS analysis.…”

Section: Sample Preparation Approachesmentioning

confidence: 99%

“…42 An example of such technique is Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA), also referred to as immunoselected reaction monitoring technology. 43 This approach allows reduction of matrix complexity within a whole-cell digest by enriching only the desired peptides, followed by MS analysis. Immuno-enrichment is not inherently capable of distinguishing between intracellular and plasma membrane proteins and adding a biotinylation step to this method could theoretically combine the advantages of the two techniques.…”

Section: Sample Quality and Characteristicsmentioning

confidence: 99%

Toward a Consensus on Applying Quantitative Liquid Chromatography‐Tandem Mass Spectrometry Proteomics in Translational Pharmacology Research: A White Paper

Prasad

Achour

Artursson

et al. 2019

Clin Pharma and Therapeutics

129

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Quantitative translation of information on drug absorption, disposition, receptor engagement, and drug–drug interactions from bench to bedside requires models informed by physiological parameters that link in vitro studies to in vivo outcomes. To predict in vivo outcomes, biochemical data from experimental systems are routinely scaled using protein quantity in these systems and relevant tissues. Although several laboratories have generated useful quantitative proteomic data using state‐of‐the‐art mass spectrometry, no harmonized guidelines exit for sample analysis and data integration to in vivo translation practices. To address this gap, a workshop was held on September 27 and 28, 2018, in Cambridge, MA, with 100 experts attending from academia, the pharmaceutical industry, and regulators. Various aspects of quantitative proteomics and its applications in translational pharmacology were debated. A summary of discussions and best practices identified by this expert panel are presented in this “White Paper” alongside unresolved issues that were outlined for future debates.

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“…To fill this gap we sought to develop a practical approach to obtain precise measurements of blood biomarkers at high frequency and low cost, using samples that can easily be collected by subjects themselves at any time and any place: dried blood microsamples. In the work described here we demonstrate the use of such a method measuring pre-selected health-relevant proteins by SISCAPA immuno-mass spectrometry [5] in conventional dried blood spot (DBS) samples collected on filter paper cards over spans of years, including extensive periods of daily collection. While this approach could be considered excessive in the context of most routine diagnostic applications, it has enormous potential value when applied to specific periods of time during which significant health events are anticipated: e.g., disease progression studies, clinical trials, surgery, or pre-arranged changes in medication, lifestyle or diet [6].…”

Section: Introductionmentioning

confidence: 99%

Multiplexed measurement of protein biomarkers in high-frequency longitudinal dried blood spot (DBS) samples: characterization of inflammatory responses

Nl¹,

Razavi²,

Me³

et al. 2019

Preprint

Self Cite

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A detailed understanding of changes in blood protein biomarkers occuring in individuals over time would enable truly personalized approaches to health and disease monitoring. Such measurements could reveal smaller, earlier departures from normal baseline levels of biomarkers thus allowing better disease detection and treatment monitoring. Current practice, however, generally involves infrequent, sporadic biomarker testing, and this undersampling likely fails to capture important biological phenomena. Here we report the use of a robust multiplex immunomass spectrometric method (SISCAPA) to measure a panel of clinically-relevant proteins in a unique collection of 1,522 dried blood spots collected longitudinally by 8 individuals over periods of up to 9 years, with daily sampling during some intervals. Analytical workflow CVs of 2-6% for most assays were achieved by normalizing DBS plasma volume using a set of 3 minimally varying proteins, facilitating temporal analysis of both high-and low-amplitude biomarker changes compared to personalized baselines. The biomarkers included a panel of 9 positive and 5 negative acute phase response (inflammatory) proteins, allowing longitudinal analysis of inflammation markers associated with major and minor infections, influenza vaccination, recovery from hip-replacement surgery and Crohn's disease. The results illustrate complex time-dependent "biomarker trajectories" on multiple timescales and provide a basis for detailed personalized models of inflammation dynamics. The striking stability of most biomarker protein levels over time, combined with the convenience of self-sampling and low cost of multiplexed measurements using mass spectrometry, provide a new window into the temporal dynamics of disease processes. The extensive results obtained using this high throughput approach offer a new source of precision biomarker 'big data' amenable to machine learning approaches and application to more personalized health monitoring.

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High precision quantification of human plasma proteins using the automated SISCAPA Immuno-MS workflow

Cited by 61 publications

References 22 publications

Clinical translation of MS-based, quantitative plasma proteomics: status, challenges, requirements, and potential

Clinical translation of MS-based, quantitative plasma proteomics: status, challenges, requirements, and potential

Toward a Consensus on Applying Quantitative Liquid Chromatography‐Tandem Mass Spectrometry Proteomics in Translational Pharmacology Research: A White Paper

Multiplexed measurement of protein biomarkers in high-frequency longitudinal dried blood spot (DBS) samples: characterization of inflammatory responses

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