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Pre-lab proteolysis for dried serum spots – a paper-based sampling concept targeting low abundant biomarkers

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Abstract

Paper-based sampling of biological matrices in combination with mass spectrometry has proven to be a promising technique for bottom-up analysis of proteins. However, these techniques have been challenging for determining low abundance proteins (pg mL−1 to low ng mL−1) due to low sampling volumes and adsorption of proteins to the sampling material. Here we are introducing a paper-based sampling concept in chip format with in-device proteolysis combined with selective proteotypic epitope peptide extraction. The sampling concept is demonstrated with the low abundant biomarker proGRP and showed promising performance from 500 pg mL−1 to 1000 ng mL−1 (R2 = 0.99) of proGRP spiked to human serum samples (all RSDs within 26%) in a five point concentration curve (n = 3). LOD and LOQ were determined to 150 pg mL−1 and 500 pg mL−1, respectively. The combination of paper-based sampling with in-device proteolysis and subsequent proteotypic epitope peptide immunocapture showed promising performance for other clinical applications.

Graphical abstract: Pre-lab proteolysis for dried serum spots – a paper-based sampling concept targeting low abundant biomarkers

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Publication details

The article was received on 11 Sep 2019, accepted on 26 Nov 2019 and first published on 29 Nov 2019


Article type: Paper
DOI: 10.1039/C9AY01976F
Anal. Methods, 2019, Advance Article

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    Pre-lab proteolysis for dried serum spots – a paper-based sampling concept targeting low abundant biomarkers

    Ø. Skjærvø, T. G. Halvorsen and L. Reubsaet, Anal. Methods, 2019, Advance Article , DOI: 10.1039/C9AY01976F

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