A Nanobioarray Chip Intended for Fast and Multiplex Antibody Detection in Biological Fluids Delivered by Centrifugal Pumping

A nanobioarray (NBA) chip has been developed with the goal of having a high throughput system that analyze proteins of low sample volumes against multiple probes in a short time. A combination of horizontal and vertical channels are produced to create an antigen array on the surface of the NBA chip in one dimension that is probed by flowing protein samples (e.g. antibodies) from biological fluids in the orthogonal dimension. To improve sensitivity, we have tested the NBA chip by immobilizing streptavidin and then biotinylated peptide to detect the presence of a mouse monoclonal antibody (MAb) that is specific for the peptide. Bound antibody is detected by an AlexaFluor 647 labeled goat (anti-mouse IgG) polyclonal antibody. Using this NBA chip, we have successfully detected antibodies in samples in 500 nL containing 50 pM of MAb (or 25 attomoles), and this demonstrates a detection limit below that of a comparable ELISA, utilizing a shorter reaction time. Such a system is intended for assays of protein mixtures in biofluids, even for proteomic studies.