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Method Article
Sandeep Kumar Vashist
Laboratory for MEMS Applications, Department of Microsystems Engineering -IMTEK, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany; HSG-IMIT - Institut für Mikro- und Informationstechnik, Georges-Koehler-Allee 103, 79110 Freiburg,
Corresponding Author
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This work is licensed under a CC BY-NC 3.0 License. Read Full License
A highly-sensitive sandwich enzyme-linked immunosorbent (ELISA) procedure was developed for detecting human fetuin A (HFA), a specific biomarker for a plethora of various infection and inflammation associated diseases. In this one-step antibody (Ab) immobilization, the capture anti-HFA Ab formed a stable complex with 3-aminopropyltriethoxysilane (APTES), via ionic and hydrophobic interactions. The complex adsorbed on a microtiter plate (MTP) as an ELISA platform for HFA, and exhibited linearity from 4.9 pg mL-1 to 20 ng mL-1, a limit of detection of 7 pg mL-1 and an analytical sensitivity of 10 pg mL-1, i.e. 51-fold more sensitive than the commercial sandwich ELISA kit. The precision obtained by this procedure for the detection of HFA spiked in human whole blood and plasma was in agreement with the commercial kit. The anti-HFA Ab-bound MTPs stored at 4°C in 0.1 M phosphate-buffered saline, pH 7.4 displayed no significant decrease in original activities after two months.
Keywords
ELISA, human fetuin A, 3-aminopropyltriethoxysilane (APTES), one-step antibody immobilization, in vitro diagnostics.
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The authors declare no competing financial interests.
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Associated Publications
One-step antibody immobilization-based rapid and highly-sensitive sandwich ELISA procedure for potential in vitro diagnostics
by Sandeep Kumar Vashist, E. Marion Schneider, Edmond Lam, +2
Scientific Reports (18 March, 2014)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Sandeep Kumar Vashist
Laboratory for MEMS Applications, Department of Microsystems Engineering -IMTEK, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany; HSG-IMIT - Institut für Mikro- und Informationstechnik, Georges-Koehler-Allee 103, 79110 Freiburg,
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 20 Mar, 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 highly-sensitive sandwich enzyme-linked immunosorbent (ELISA) procedure was developed for detecting human fetuin A (HFA), a specific biomarker for a plethora of various infection and inflammation associated diseases. In this one-step antibody (Ab) immobilization, the capture anti-HFA Ab formed a stable complex with 3-aminopropyltriethoxysilane (APTES), via ionic and hydrophobic interactions. The complex adsorbed on a microtiter plate (MTP) as an ELISA platform for HFA, and exhibited linearity from 4.9 pg mL-1 to 20 ng mL-1, a limit of detection of 7 pg mL-1 and an analytical sensitivity of 10 pg mL-1, i.e. 51-fold more sensitive than the commercial sandwich ELISA kit. The precision obtained by this procedure for the detection of HFA spiked in human whole blood and plasma was in agreement with the commercial kit. The anti-HFA Ab-bound MTPs stored at 4°C in 0.1 M phosphate-buffered saline, pH 7.4 displayed no significant decrease in original activities after two months.
Figure 1
Figure 2
Figure 3
Figure 4
The authors declare no competing financial interests.
Loading...
Associated Publications
One-step antibody immobilization-based rapid and highly-sensitive sandwich ELISA procedure for potential in vitro diagnostics
by Sandeep Kumar Vashist, E. Marion Schneider, Edmond Lam, +2
Scientific Reports (18 March, 2014)