A highly-sensitive sandwich ELISA procedure based on one-step antibody immobilization for potential in vitro diagnostics
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.
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Posted 20 Mar, 2014
A highly-sensitive sandwich ELISA procedure based on one-step antibody immobilization for potential in vitro diagnostics
Posted 20 Mar, 2014
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.
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