Development of a cost-effective multiplexed antibody microarray for qualitative and quantitative detection of multi-analytes

Using a multi-analyte detection method instead of a single analyte detection method can provide more information for an investigation with higher accuracy and reliability. Currently developed multi-analyte rapid immunoassays provide convenient and rapid qualitative and quantitative assays. However, these test formats cannot achieve high-throughput measurements. Multiplexed antibody microarrays in planar and on microspheres in suspension are also common multi-analyte immunoassay formats. They support relatively more analyte detection than a single assay. However, these types of immunoassays are mainly based on fluorescence, chemiluminescence and electrochemical methods which are relatively complicated to handle, time consuming and heavily reliant imaging systems and analyzers for signal quantification. These test formats do not provide an instant naked eye observable signal so they are not able to provide a qualitative measurement for convenient use. In view of this, a multiplexed antibody microarray for qualitative and quantitative detection of multi-analytes was developed. The technology is based on sandwich immunoassay principle with colloidal gold as signaling label for detection in a 96-well polyvinylidene fluoride filter plate. The feasibility of this technology has been also illustrated by developing a heart attack panel for detection of heart type fatty acid binding protein (H-FABP) and cardiac troponin I (cTnI) to diagnose acute myocardial infarction (AMI) and a heart risk panel for detection of C-reactive protein (CRP) and myeloperoxidase (MPO) to predict risk of cardiovascular disease