Abstract
In this paper, we present the ImmunoDisk, a fully automated sample-to-answer centrifugal microfluidic cartridge, integrating a heterogeneous, wash-free, magnetic- and fluorescent bead-based immunoassay (bound-free phase detection immunoassay/BFPD-IA). The BFPD-IA allows the implementation of a simple fluidic structure, where the assay incubation, bead separation and detection are performed in the same chamber. The system was characterized using a C-reactive protein (CRP) competitive immunoassay. A parametric investigation on air drying of protein-coupled beads for pre-storage at room temperature is presented. The key parameters were buffer composition, drying temperature and duration. A protocol for drying two different types of protein-coupled beads with the same temperature and duration using different drying buffers is presented. The sample-to-answer workflow was demonstrated measuring CRP in 5 µL of human serum, without prior dilution, utilizing only one incubation step, in 20 min turnaround time, in the clinically relevant concentration range of 15–115 mg/L. A reproducibility assessment over three disk batches revealed an average signal coefficient of variation (CV) of 5.8 ± 1.3%. A CRP certified reference material was used for method verification with a concentration CV of 8.6%. Our results encourage future testing of the CRP-ImmunoDisk in clinical studies and its point-of-care implementation in many diagnostic applications.
Funder
European Commission
Eurostars, managed by the Federal Ministry of Education & Research
Subject
Clinical Biochemistry,General Medicine,Analytical Chemistry,Biotechnology,Instrumentation,Biomedical Engineering,Engineering (miscellaneous)
Reference80 articles.
1. Handbook of Immunoassay Technologies: Approaches, Performances, and Applications,2018
2. Salivary biomarkers detection: Analytical and immunological methods overview
3. Label‐Free Quantitative Proteomics versus Antibody‐Based Assays to Measure Neutrophil‐Derived Enzymes in Saliva
4. ELISA: Methods and Protocols;Hnasko,2015
5. Immunoassays: Development, Applications and Future Trends;O’Kennedy,2017
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