There is a need for low-cost immunoassays that measure the presence and concentration of multiple harmful agents in one device. Currently, comparable immunoassays employ a one-analyte-per-test format that is time consuming and not cost effective for the requirement of detecting multiple analytes in a single sample. For instance, if a spectrum of harmful agents, including E. coli O157, cholera toxin, and Salmonella typhimurium, should be simultaneously monitored in foods and drinking water, then a one-analyte-per-test would be inefficient. This work demonstrates a platform capable of simultaneous detection of multiple analytes in a single, low-cost, microvalve array-enabled multiplexed immunoassay. This multiplexed immunoassay platform is demonstrated in a prototype COC (cyclic olefin copolymer) device with a 2×3 array in which 6 analytes can be detected simultaneously. In order to contain and regulate the flow of reagents in the multichannel device, an array of microfluidic valves actuated by a thermally expandable material and microfabricated resistors have been developed to direct the flow to the necessary assay sites. The microvalve-based immunoassay is shown to be reliable, easy to operate, and compatible with large-scale integration. The all-plastic microvalves use paraffin wax as the thermally sensitive material which drastically reduces power consumption by latching upon closing so that pulsed power is required only to close and latch the microvalve until it is necessary to re-open the valve. The multiplexed detection scheme has been demonstrated by using three proteins, C reactive protein (CRP) and transferrin, both of which are biomarkers associated with traumatic brain injury (TBI) as well as bovine serum albumin (BSA) as the negative control. Since there are no external bulky pneumatic accessories required to operate/latch the microvalves in the device, this compact, thermally actuated and latching microvalve-enabled multiplexed immunoassay has the potential to realize a portable, low power, battery operated microfluidic device for biological assays.

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