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@article{1479457,
author = {Vidová, Veronika and Stuchlíková, Eliška and Vrbová, Markéta and Almáši, Martina and Klánová, Jana and Thon, Vojtěch and Spáčil, Zdeněk},
article_location = {Washington, USA},
article_number = {1},
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},
language = {eng},
issn = {1535-3893},
journal = {Journal of Proteome Research},
title = {Multiplex Assay for Quantification of Acute Phase Proteins and Immunoglobulin A in Dried Blood Spots},
url = {https://pubs.acs.org/doi/10.1021/acs.jproteome.8b00657},
volume = {18},
year = {2019}
}
TY - JOUR
ID - 1479457
AU - Vidová, Veronika - Stuchlíková, Eliška - Vrbová, Markéta - Almáši, Martina - Klánová, Jana - Thon, Vojtěch - Spáčil, Zdeněk
PY - 2019
TI - Multiplex Assay for Quantification of Acute Phase Proteins and Immunoglobulin A in Dried Blood Spots
JF - Journal of Proteome Research
VL - 18
IS - 1
SP - 380-391
EP - 380-391
PB - AMER CHEMICAL SOC
SN - 15353893
KW - inflammation markers
KW - acute phase proteins
KW - immune response
KW - dried blood spots
KW - targeted quantitative proteomics
KW - selected reaction monitoring
UR - https://pubs.acs.org/doi/10.1021/acs.jproteome.8b00657
L2 - https://pubs.acs.org/doi/10.1021/acs.jproteome.8b00657
N2 - 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).
ER -
VIDOVÁ, Veronika, Eliška STUCHLÍKOVÁ, Markéta VRBOVÁ, Martina ALMÁŠI, Jana KLÁNOVÁ, Vojtěch THON a Zdeněk SPÁČIL. Multiplex Assay for Quantification of Acute Phase Proteins and Immunoglobulin A in Dried Blood Spots. \textit{Journal of Proteome Research}. Washington, USA: AMER CHEMICAL SOC, 2019, roč.~18, č.~1, s.~380-391. ISSN~1535-3893. Dostupné z: https://dx.doi.org/10.1021/acs.jproteome.8b00657.
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