Authors
Sandeep Kumar Vashist, E. Marion Schneider, Edmond Lam, Sabahudin Hrapovic & John H.T. Luong
Summary
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.Further details
The protocol was published on Protocol Exchange on 20 March 2014. To see the entire protocol, click on the source link.Advertisement
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