Authors
Sandeep Kumar Vashist
Summary
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.
Further details
The protocol was published on Protocol Exchange on 31 December 2013. To see the entire protocol, click on the source link.
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