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.

Basic information

Original name

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

Authors

JOHANNSEN, B. (guarantor), Michal KARPÍŠEK (203 Czech Republic, belonging to the institution), D. BAUMGARTNER, V. KLEIN, N. BOSTANCI, N. PAUST, S.M. FRUEH, R. ZENGERLE and K. MITSAKAKIS

Edition

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

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30104 Pharmacology and pharmacy

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 6.911

RIV identification code

RIV/00216224:14160/21:00122556

Organization unit

Faculty of Pharmacy

DOI

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

UT WoS

000635606500009

Keywords in English

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

Abstract

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 and K. MITSAKAKIS. One-step, wash-free, bead-based immunoassay employing bound-free phase detection. Analytica Chimica Acta. Amsterdam: Elsevier, 2021, vol. 1153, No 1, p. 1-9. ISSN 0003-2670. Available from: 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 and K. MITSAKAKIS. One-step, wash-free, bead-based immunoassay employing bound-free phase detection. \textit{Analytica Chimica Acta}. Amsterdam: Elsevier, 2021, vol.~1153, No~1, p.~1-9. ISSN~0003-2670. Available from: https://dx.doi.org/10.1016/j.aca.2021.338280.

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