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Method Article
Sandeep Kumar Vashist
Department of Microsystems Engineering (IMTEK), University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany.
Corresponding Author
License:
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We report a highly-sensitive graphene nanoplatelets (GNPs)-based sandwich enzyme-linked immunosorbent immunoassay (ELISA) procedure for the detection of human lipocalin-2 (LCN2). It involves the functionalization of 96-well microtiter plate (MTP) with GNPs (dispersed in 3-aminopropyltriethoxysilane (APTES)). APTES generates the free amino groups on MTP and GNPs, which are used for the leach-proof covalent binding of anti-LCN2 capture antibody by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) based heterobifunctional crosslinking. The developed immunoassay detects LCN2 in the range of 0.6-5120 pg mL-1 with limit of detection and analytical sensitivity of 0.7 and 1 pg mL-1, respectively. It is the most sensitive immunoassay for LCN2, which has 80-fold higher analytical sensitivity and 3-fold reduced immunoassay duration than the commercial sandwich ELISA kit. The anti-LCN2 antibody-bound MTPs, stored at 4°C in 0.1M phosphate buffered saline (PBS), were highly stable as they did not show any decrease in functional activity for 8 weeks.
Keywords
Sandwich ELISA, human lipocalin-2, graphene nanoplatelets, EDC, APTES
Figure 1
Figure 2
The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.docx
? Troubleshooting Troubleshooting Table TABLE 1. Troubleshooting table.
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Associated Publications
Graphene-based immunoassay for human lipocalin-2
by Sandeep Kumar Vashist
Analytical Biochemistry (30 December, 2013)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Sandeep Kumar Vashist
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 31 Dec, 2013
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
We report a highly-sensitive graphene nanoplatelets (GNPs)-based sandwich enzyme-linked immunosorbent immunoassay (ELISA) procedure for the detection of human lipocalin-2 (LCN2). It involves the functionalization of 96-well microtiter plate (MTP) with GNPs (dispersed in 3-aminopropyltriethoxysilane (APTES)). APTES generates the free amino groups on MTP and GNPs, which are used for the leach-proof covalent binding of anti-LCN2 capture antibody by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) based heterobifunctional crosslinking. The developed immunoassay detects LCN2 in the range of 0.6-5120 pg mL-1 with limit of detection and analytical sensitivity of 0.7 and 1 pg mL-1, respectively. It is the most sensitive immunoassay for LCN2, which has 80-fold higher analytical sensitivity and 3-fold reduced immunoassay duration than the commercial sandwich ELISA kit. The anti-LCN2 antibody-bound MTPs, stored at 4°C in 0.1M phosphate buffered saline (PBS), were highly stable as they did not show any decrease in functional activity for 8 weeks.
Figure 1
Figure 2
The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.docx
? Troubleshooting Troubleshooting Table TABLE 1. Troubleshooting table.
Loading...
Associated Publications
Graphene-based immunoassay for human lipocalin-2
by Sandeep Kumar Vashist
Analytical Biochemistry (30 December, 2013)