Authors
Sandeep Kumar Vashist
Summary
We report a highly-sensitive chemiluminescent immunoassay (CIA) for the detection of human fetuin A (HFA), which is based on the leach-proof covalent crosslinking of anti-HFA capture antibodies on 3-aminopropyltriethoxysilane (APTES)-functionalized 96-well chemiluminescent microtiter plates (CMTP) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysulfosuccinimide. It has more than 3-fold reduced overall assay duration than the commercial enzyme-linked immunosorbent assay (ELISA). Moreover, it has a wide detection range i.e. 0.3 pg/mL-20 ng/mL and 125-fold better analytical sensitivity than the commercial ELISA kit. The anti-HFA antibody-bound MTPs can be effectively stored in 0.1M PBS without any significant loss of functional activity. The developed HFA CIA is multisubstrate-compatible and can be employed for immunoassays on a wide range of commercially-relevant substrates that are being used in various biosensors/point-of-care diagnostics.Further details
The protocol was published on Protocol Exchange on 24 October 2012. To see the entire protocol, click on the source link.Advertisement
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