Publication details

Multiplex Assay for Quantification of Acute Phase Proteins and Immunoglobulin A in Dried Blood Spots

Authors

VIDOVÁ Veronika STUCHLÍKOVÁ Eliška VRBOVÁ Markéta ALMÁŠI Martina KLÁNOVÁ Jana THON Vojtěch SPÁČIL Zdeněk

Year of publication 2019
Type Article in Periodical
Magazine / Source Journal of Proteome Research
MU Faculty or unit

Faculty of Science

Citation
Web Full Text
Doi http://dx.doi.org/10.1021/acs.jproteome.8b00657
Keywords inflammation markers; acute phase proteins; immune response; dried blood spots; targeted quantitative proteomics; selected reaction monitoring
Description Inflammation is the first line defense mechanism against infection, tissue damage, or cancer development. However, inappropriate inflammatory response may also trigger diseases. The quantification of inflammatory proteins is essential to distinguish between harmful and beneficial immune response. Currently used immunoanalytical assays may suffer specificity issues due to antigen-antibody interaction and possible cross-reactivity of antibody with other protein species. In addition, immunoanalytical assays typically require invasive blood sampling and additional logistics; they are relatively costly and highly challenging to multiplex. We present a multiplex assay based on selected reaction monitoring (SRM) for quantification of seven acute-phase proteins (i.e., SAA1, SAA2-isoform1, SAA4, CRP, A1AT-isoform1, A1AG1, A1AG2) and the adaptive immunity effector IGHA1 in dried blood spots. This type of sample is readily available from all human subjects including newborns. The study utilizes proteotypic isotopically labeled peptides with trypsin-cleavable tag and presents optimized and reproducible workflow and several important practical remarks regarding quantitative SRM assays development. The panel of inflammatory proteins was quantified with sequence specificity capable to differentiate protein isoforms with intra- and interday precision (<16.4% coefficient of variation (CV) and <14.3% CV, respectively). Quantitative results were correlated with immuno-nephelometric assay (typically greater than 0.9 Pearson's R).
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