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Method Article
Sandeep Kumar Vashist
HSG-IMIT - Institut für Mikro- und Informationstechnik; Laboratory for MEMS Applications, Department of Microsystems Engineering -IMTEK, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany
Corresponding Author
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A rapid one-step kinetics-based sandwich enzyme-linked immunosorbent (ELISA) procedure has been developed for highly-sensitive detection of C-reactive protein (CRP) in less than 30 min. With minimal process steps, the procedure is highly simplified and cost-effective. The analysis only involves sequentially the formation of a sandwich immune complex on capture anti-CRP antibody (Ab)-bound Dynabeads, followed by two magnet-assisted washings and an enzymatic reaction. The developed immunoassay (IA) detected CRP in the dynamic range of 0.3-81 ng mL-1 with a limit of detection and analytical sensitivity of 0.4 ng mL-1 and 0.7 ng mL-1. Its analytical precision for analysis of CRP spiked in diluted human serum, whole blood, and ethylenediaminetetraacetic acid (EDTA) plasma samples of patients was validated by conventional ELISA, unravelling its immense potential for in vitro diagnostics (IVD).
Keywords
One-step kinetics, immunoassay, C-reactive protein, Dynabeads, in vitro diagnostics
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The authors declare no competing financial interests.
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Associated Publications
One-step kinetics-based immunoassay for the highly sensitive detection of C-reactive protein in less than 30min
by Sandeep Kumar Vashist, Gregor Czilwik, Thomas van Oordt, +4
Analytical Biochemistry (23 May, 2014)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Sandeep Kumar Vashist
HSG-IMIT - Institut für Mikro- und Informationstechnik; Laboratory for MEMS Applications, Department of Microsystems Engineering -IMTEK, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 27 May, 2014
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
A rapid one-step kinetics-based sandwich enzyme-linked immunosorbent (ELISA) procedure has been developed for highly-sensitive detection of C-reactive protein (CRP) in less than 30 min. With minimal process steps, the procedure is highly simplified and cost-effective. The analysis only involves sequentially the formation of a sandwich immune complex on capture anti-CRP antibody (Ab)-bound Dynabeads, followed by two magnet-assisted washings and an enzymatic reaction. The developed immunoassay (IA) detected CRP in the dynamic range of 0.3-81 ng mL-1 with a limit of detection and analytical sensitivity of 0.4 ng mL-1 and 0.7 ng mL-1. Its analytical precision for analysis of CRP spiked in diluted human serum, whole blood, and ethylenediaminetetraacetic acid (EDTA) plasma samples of patients was validated by conventional ELISA, unravelling its immense potential for in vitro diagnostics (IVD).
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.pdf
Supplementary information Supplementary data
Loading...
Associated Publications
One-step kinetics-based immunoassay for the highly sensitive detection of C-reactive protein in less than 30min
by Sandeep Kumar Vashist, Gregor Czilwik, Thomas van Oordt, +4
Analytical Biochemistry (23 May, 2014)