High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA)

A new generation biochip is described as capable of supporting high-through put (HT), multiplexed enzyme-linked immunosorbent assays (ELISAs). These bi ochips consist of an optically flat, glass plate containing 96 wells formed by an enclosing hydrophobic Teflon(R) mask. The footprint dimensions of ea ch well and the plate precisely match those of a standard microplate. Each well contains four identical 36-element arrays (144 elements per well) comp rising 8 different antigens and a marker protein. Arrays are formed by a cu stom continuous flow, capillary-based print head attached to a precise, hig h-speed, X-Y-Z robot. The array printing capacity of a single robot exceeds 20 000 arrays per day. Arrays are quantitatively imaged using a custom, hi gh-resolution, scanning charge-coupled device (CCD) detector with an imagin g throughput of 96 arrays every 30 s. Using this new process, arrayed antig ens were individually and collectively detected using standard ELISA techni ques. Experiments demonstrate that specific multiplex detection of protein antigens arrayed on a glass substrate is feasible. Because of the open micr oarray architecture, the 96-well microarray format is compatible with autom ated robotic systems and supports a low-cost, highly parallel assay format Future applications of this new high-throughput screening (HTS)format inclu de direct cellular protein expression profiling multiplexed assays for dete ction of infectious agents and cancer diagnostics.