A1 Refereed original research article in a scientific journal

Targeted and Untargeted Amine Metabolite Quantitation in Single Cells with Isobaric Multiplexing




AuthorsHeininen, Juho; Movahedi, Parisa; Kotiaho, Tapio; Kostiainen, Risto; Pahikkala, Tapio; Teppo, Jaakko

PublisherWiley-VCH

Publication year2024

JournalChemistry - A European Journal

Journal acronymChemistry

Article numbere202403278

Volume30

Issue72

ISSN0947-6539

eISSN1521-3765

DOIhttps://doi.org/10.1002/chem.202403278

Web address https://doi.org/10.1002/chem.202403278

Self-archived copy’s web addresshttps://research.utu.fi/converis/portal/detail/Publication/459037549


Abstract

We developed a single cell amine analysis approach utilizing isobarically multiplexed samples of 6 individual cells along with analyte abundant carrier. This methodology was applied for absolute quantitation of amino acids and untargeted relative quantitation of amines in a total of 108 individual cells using nanoflow LC with high-resolution mass spectrometry. Together with individually determined cell sizes, this provides quantification of intracellular concentrations within individual cells. The targeted method was partially validated for 10 amino acids with limits of detection in low attomoles, linear calibration range covering analyte amounts typically from 30 amol to 120 fmol, and correlation coefficients (R) above 0.99. This was applied with cell sizes recorded during dispensing to determine millimolar intracellular amino acid concentrations. The untargeted approach yielded 249 features that were detected in at least 25% of the single cells, providing modest cell type separation on principal component analysis. Using Greedy forward selection with regularized least squares, a sub-selection of 100 features explaining most of the difference was determined. These features were annotated using MS2 from analyte standards and accurate mass with library search. The approach provides accessible, sensitive, and high-throughput method with the potential to be expanded also to other forms of ultrasensitive analysis.


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Funding information in the publication
Research Council of Finland (project #321472) is acknowledged for funding.


Last updated on 2025-24-02 at 13:09