This technology includes the enablement of the nanoliter-scale transfer of biological liquids in array format from a microplate (source plate) containing cultured cells or other protein-containing mixtures onto a nitrocellulose membrane that has been mounted within a custom-designed target plate. Using this method and the prototype nitrocellulose target plate, reverse phase protein arrays can be generated in which protein levels from each well transferred onto the membrane can be detected and quantified. Various designs for the nitrocellulose target plate are in development to offer flexibility into the immunoblotting process for both manual and automation compatibility. Importantly, this process is 384-well and 1536-well compatible to enable its use in high-throughput screening, and is capable of detecting native, endogenous proteins depending on the antibodies used. The discovery connects two seemingly disparate biological concepts, acoustic dispensing and immunoblotting, to enable protein-based high-throughput screening among other applications while minimizing cost, reagents, and biological material.
The present discovery enables the generation of reverse phase protein arrays for the detection of native, untagged proteins in biological liquids (cell culture media, cell lysate, and other protein-containing mixtures) in 384-well and 1536-well formats, for which there are currently very few available options.