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.
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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
Original language English
Type of outcome Article in a journal
Field of Study 30104 Pharmacology and pharmacy
Country of publisher Netherlands
Confidentiality degree is not subject to a state or trade secret
WWW 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
Tags rivok, ÚFTo
Tags International impact, Reviewed
Changed by Changed by: JUDr. Sabina Krejčiříková, učo 383857. Changed: 14/4/2022 14:06.
Abstract
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.
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