Detection of multiple proteins in an antibody-based protein microarray system

A new antibody-based protein array assay is described. This assay combines the advantages of the specificity of enzyme-linked immunosorbent assays (EL ISA), sensitivity of enhanced chemiluminescence (ECL) and high-throughput o f microspot. In this system, the capture proteins, either antibodies or ant igens are spotted onto membranes in an array format. Biological samples are then incubated with membranes. After antigens or antibodies in the samples bind to their corresponding targets and unbound proteins are washed away, the membranes are exposed to Horseradish Peroxidase (HRP)-conjugated antibo dy(ies). The signals are finally visualized with ECL system. Experiments de monstrate that multiple cytokines and antibodies can be simultaneously dete cted using this new approach. The procedure is so simple that no sophistica ted equipment is required. The concept should be able to be extended to dev elop a high-throughput protein array system. Future applications of this ne w approach include direct protein expression profiling, immunological disea se diagnostics and discovery of new biomarkers. (C) 2001 Elsevier Science B .V. All rights reserved.