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- Title
Development and performance of NLISA for C-reactive protein detection based on Prussian blue nanoparticle conjugates.
- Authors
Nikitina, Maria; Khramtsov, Pavel; Bochkova, Maria; Rayev, Mikhail
- Abstract
Prussian blue nanoparticles (PBNPs), also called nanozymes, are very attractive as an alternative to horseradish peroxidase in immunoassay development due to their simple and low-cost synthesis, stability and high catalytic activity. Today, there is a method for highly effective PBNP synthesis based on the reduction of an FeCl3/K3[Fe(CN)6] mixture by hydrogen peroxide. However, there is a lack of research showcasing the use of these highly effective PBNPs for specific target detection in clinical settings, as well as a lack of comprehensive comparisons with conventional methods. To address this gap, we prepared diagnostic reagents based on highly effective PBNPs by modifying them using gelatin and attaching anti-C-reactive protein (CRP) monoclonal antibodies through cross-linking with glutaraldehyde. As a result, a solid-phase colorimetric immunoassay in a sandwich format (nanozyme-linked immunosorbent assay [NLISA]) using highly effective PBNPs as a label for CRP detection has been demonstrated for the first time. The assay demonstrated a detection limit of 21.8 pg/mL, along with acceptable selectivity, precision (CV < 25%) and accuracy (the recovery index was within acceptable limits (75–125%) for LLOQ /ULOQ range. The analytical performance of this method is on par with sensitive assays developed in the last 5 years. Notably, the results obtained from NLISA align with those from an immunofluorescence assay conducted by a certified clinical laboratory. Furthermore, this study underscores the technological challenges involved in constructing an analysis that necessitate further exploration.
- Subjects
C-reactive protein; NANOPARTICLES; PRUSSIAN blue; HORSERADISH peroxidase; HYDROGEN peroxide; SYNTHETIC enzymes; PEROXIDASE; GELATIN
- Publication
Analytical & Bioanalytical Chemistry, 2024, Vol 416, Issue 13, p3097
- ISSN
1618-2642
- Publication type
Article
- DOI
10.1007/s00216-024-05268-y