One-step, wash-free, bead-based immunoassay employing
bound-free phase detection.

J 2021

One-step, wash-free, bead-based immunoassay employing bound-free phase detection.

JOHANNSEN, B., Michal KARPÍŠEK, D. BAUMGARTNER, V. KLEIN, N. BOSTANCI et. al.

Základní údaje

Originální název

One-step, wash-free, bead-based immunoassay employing bound-free phase detection.

Autoři

JOHANNSEN, B. (garant), Michal KARPÍŠEK (203 Česká republika, domácí), D. BAUMGARTNER, V. KLEIN, N. BOSTANCI, N. PAUST, S.M. FRUEH, R. ZENGERLE a K. MITSAKAKIS

Vydání

Analytica Chimica Acta, Amsterdam, Elsevier, 2021, 0003-2670

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30104 Pharmacology and pharmacy

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 6.911

Kód RIV

RIV/00216224:14160/21:00122556

Organizační jednotka

Farmaceutická fakulta

DOI

http://dx.doi.org/10.1016/j.aca.2021.338280

UT WoS

000635606500009

Klíčová slova anglicky

Bound-free phase; Immunoassay; Inflammation; Micro/nanoparticles; Oral biomarkers; Saliva

Štítky

rivok, ÚFTo

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 14. 4. 2022 14:06, JUDr. Sabina Krejčiříková

Anotace

V originále

We present a simple and fast one-step heterogeneous immunoassay, with performance characteristics that can enable easy and versatile adaptation to miniaturized, automated point-of-care systems. This novel analytical method uses magnetic and fluorescent beads as capture and detection agents respectively. Its main feature is the measurement of the fluorescent signal in the bound-free phase for (semi-)quantitative detection of analytes. Thus, no washing is required and the workflow consists only of sample and reagent supply, incubation, separation and detection. The immunoassay concept is demonstrated with C-reactive protein (CRP), a systemic inflammation marker. CRP in only 5 μL of undiluted serum was measured in the range 20-140 mg L-1 (includes clinically relevant cut-off values). The limit of detection (LOD) was 22.1 ± 6.3 mg L-1 (incubation 15 min). A CRP certified reference material was measured on five different days. Intra- and inter-assay coefficients of variation were 4.6 ± 1.9% and 5.6% respectively. To demonstrate the compatibility of the assay concept with additional matrices and concentration ranges, three oral inflammation markers, namely matrix metalloproteinases 8 and 9 (MMP-8, MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1), were measured in saliva in the ranges 0.47-30 ng mL-1 for MMP-8 and MMP-9, and 0.69-44 ng mL-1 for TIMP-1. LODs were 0.24 ng mL-1, 0.38 ng mL-1 and 0.39 ng mL-1 respectively (incubation 20 min). Multiplexing capacity of the assay concept was also shown with these markers. The demonstrated excellent reproducibility of the results, combined with the versatility and low complexity of the introduced immunoassay concept, make it an attractive candidate for applied analytical chemistry and automated point-of-care testing.

Citovat

JOHANNSEN, B., Michal KARPÍŠEK, D. BAUMGARTNER, V. KLEIN, N. BOSTANCI, N. PAUST, S.M. FRUEH, R. ZENGERLE a K. MITSAKAKIS. One-step, wash-free, bead-based immunoassay employing bound-free phase detection. Analytica Chimica Acta. Amsterdam: Elsevier, 2021, roč. 1153, č. 1, s. 1-9. ISSN 0003-2670. Dostupné z: https://dx.doi.org/10.1016/j.aca.2021.338280.

@article{1797205,
   author = {Johannsen, B. and Karpíšek, Michal and Baumgartner, D. and Klein, V. and Bostanci, N. and Paust, N. and Frueh, S.M. and Zengerle, R. and Mitsakakis, K.},
   article_location = {Amsterdam},
   article_number = {1},
   doi = {http://dx.doi.org/10.1016/j.aca.2021.338280},
   keywords = {Bound-free phase; Immunoassay; Inflammation; Micro/nanoparticles; Oral biomarkers; Saliva},
   language = {eng},
   issn = {0003-2670},
   journal = {Analytica Chimica Acta},
   title = {One-step, wash-free, bead-based immunoassay employing bound-free phase detection.},
   url = {https://www.sciencedirect.com/science/article/pii/S0003267021001069?via%3Dihub},
   volume = {1153},
   year = {2021}
}

TY  - JOUR
ID  - 1797205
AU  - Johannsen, B. - Karpíšek, Michal - Baumgartner, D. - Klein, V. - Bostanci, N. - Paust, N. - Frueh, S.M. - Zengerle, R. - Mitsakakis, K.
PY  - 2021
TI  - One-step, wash-free, bead-based immunoassay employing bound-free phase detection.
JF  - Analytica Chimica Acta
VL  - 1153
IS  - 1
SP  - 1-9
EP  - 1-9
PB  - Elsevier
SN  - 00032670
KW  - Bound-free phase
KW  - Immunoassay
KW  - Inflammation
KW  - Micro/nanoparticles
KW  - Oral biomarkers
KW  - Saliva
UR  - https://www.sciencedirect.com/science/article/pii/S0003267021001069?via%3Dihub
N2  - We present a simple and fast one-step heterogeneous immunoassay, with performance characteristics that can enable easy and versatile adaptation to miniaturized, automated point-of-care systems. This novel analytical method uses magnetic and fluorescent beads as capture and detection agents respectively. Its main feature is the measurement of the fluorescent signal in the bound-free phase for (semi-)quantitative detection of analytes. Thus, no washing is required and the workflow consists only of sample and reagent supply, incubation, separation and detection. The immunoassay concept is demonstrated with C-reactive protein (CRP), a systemic inflammation marker. CRP in only 5 μL of undiluted serum was measured in the range 20-140 mg L-1 (includes clinically relevant cut-off values). The limit of detection (LOD) was 22.1 ± 6.3 mg L-1 (incubation 15 min). A CRP certified reference material was measured on five different days. Intra- and inter-assay coefficients of variation were 4.6 ± 1.9% and 5.6% respectively. To demonstrate the compatibility of the assay concept with additional matrices and concentration ranges, three oral inflammation markers, namely matrix metalloproteinases 8 and 9 (MMP-8, MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1), were measured in saliva in the ranges 0.47-30 ng mL-1 for MMP-8 and MMP-9, and 0.69-44 ng mL-1 for TIMP-1. LODs were 0.24 ng mL-1, 0.38 ng mL-1 and 0.39 ng mL-1 respectively (incubation 20 min). Multiplexing capacity of the assay concept was also shown with these markers. The demonstrated excellent reproducibility of the results, combined with the versatility and low complexity of the introduced immunoassay concept, make it an attractive candidate for applied analytical chemistry and automated point-of-care testing.
ER  -

JOHANNSEN, B., Michal KARPÍŠEK, D. BAUMGARTNER, V. KLEIN, N. BOSTANCI, N. PAUST, S.M. FRUEH, R. ZENGERLE a K. MITSAKAKIS. One-step, wash-free, bead-based immunoassay employing bound-free phase detection. \textit{Analytica Chimica Acta}. Amsterdam: Elsevier, 2021, roč.~1153, č.~1, s.~1-9. ISSN~0003-2670. Dostupné z: https://dx.doi.org/10.1016/j.aca.2021.338280.

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