Authors
Pradip Nahar & Shahila Parween
Summary
In this protocol, we report spectrophotometer-free image-based miniaturized ELISA on an activated polypropylene microtest plate (APPµTP) which is precise, reliable, rapid, and sensitive. Activated test zone in APPµTP binds a capture biomolecule through covalent linkage thereby, eliminating non-specific binding often prevalent in absorption based techniques. Efficacy of APPµTP is demonstrated by detecting human immunoglobulin G (IgG), human immunoglobulin E (IgE) and Aspergillus fumigatus antibody in patient’s sera. Detection is done by taking the image of the assay solution by a desktop scanner and analyzing the color of the image. Human IgE quantification by color saturation in the image-based assay shows excellent correlation with absorbance-based assay. APPµTP can quantify an analyte as precisely as in microtiter plate and could be a good substitute for conventional immunoassay procedure widely used in clinical and research laboratories.Further details
The protocol was published on Protocol Exchange on 31 December 2013. To see the entire protocol, click on the source link.Advertisement
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