On-line duplex molecularly imprinted solid-phase extraction for analysis of low abundant biomarkers in human serum by liquid chromatography-tandem mass spectrometry

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McKitterick, Nicholas and Bicak, Tugrul Cem and Switnicka-Plak, Magdalena A. and Cormack, Peter A.G. and Reubsaet, Léon and Grønhaug Halvorsen, Trine (2021) On-line duplex molecularly imprinted solid-phase extraction for analysis of low abundant biomarkers in human serum by liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 1655. 462490. ISSN 0021-9673 (https://doi.org/10.1016/j.chroma.2021.462490)

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Abstract

In the present work, a pair of molecularly imprinted polymers (MIPs) targeting distinct peptide targets were packed into trap columns and combined for automated duplex analysis of two low abundant small cell lung cancer biomarkers (neuron-specific enolase [NSE] and progastrin-releasing peptide [ProGRP]). Optimization of the on-line molecularly imprinted solid-phase extraction (MISPE) protocol ensured that the MIPs had the necessary affinity and selectivity towards their respective signature peptide targets – NLLGLIEAK (ProGRP) and ELPLYR (NSE) – in serum. Two duplex formats were evaluated: a physical mixture of the two MIPs (1:1 w/w ratio) inside a single trap column, and two separate MIP trap columns connected in series. Both duplex formats enabled the extraction of the peptides from serum. However, the trap columns in series gave superior extraction efficiency (85.8±3.8% and 49.1±6.7% for NLLGLIEAK and ELPLYR, respectively). The optimized protocol showed satisfactory intraday (RSD≤23.4 %) and interday (RSD≤14.6%) precision. Duplex analysis of NSE and ProGRP spiked into digested human serum was linear (R 2≥0.98) over the disease range (0.3-30 nM). The estimated limit of detection (LOD) and limit of quantification (LOQ) were 0.11 nM and 0.37 nM, respectively, for NSE, and 0.06 nM and 0.2 nM, respectively, for ProGRP. Both biomarkers were determined at clinically relevant levels. To the best of our knowledge, the present work is the first report of an automated MIP duplex biomarker analysis. It represents a proof of concept for clinically viable duplex analysis of low abundant biomarkers present in human serum or other biofluids.

ORCID iDs

McKitterick, Nicholas, Bicak, Tugrul Cem, Switnicka-Plak, Magdalena A. ORCID logoORCID: https://orcid.org/0000-0003-2048-0745, Cormack, Peter A.G. ORCID logoORCID: https://orcid.org/0000-0002-3390-8176, Reubsaet, Léon and Grønhaug Halvorsen, Trine;
  • Item type:Article
    ID code:77814
    Dates:
    Date
    Event
    11 October 2021
    Published
    31 August 2021
    Published Online
    20 August 2021
    Accepted
    Subjects:Science > Chemistry
    Department:Faculty of Science > Pure and Applied Chemistry
    Depositing user: Pure Administrator
    Date deposited:16 Sep 2021 16:03
    Last modified:13 Nov 2024 23:53
    URI:https://strathprints.strath.ac.uk/id/eprint/77814
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