There is a need to create an easily deployable and point-of-care (POC) diagnostic platform for disease outbreaks and for monitoring and maintenance of chronic illnesses. Such platforms are useful in regions where access to clinical laboratories may be limited or constrained using cost-effective solutions to quickly process high numbers of samples. Using oil and water liquid-liquid interphase separation, immunoassays developed for microfluidic chips can potentially meet this need when leveraged with electromagnetic actuation and antibody-coated superparamagnetic beads. We have developed a microfluidic immunoassay detection platform, which enables assay automation and maintains successful liquid containment for future use in the field. The assay was studied through a series of magnetic and fluid simulations to demonstrate these optimizations, and an optimized chip was tested using a target model for HIV-1, the p24 capsid antigen. The use of minimal reagents further lowers the cost of each assay and lowers the required sample volume for testing (<50 μL), that can offer easy turnaround for sample collection and assay results. The developed microfluidic immunoassay platform can be easily scaled for multiplex or multi-panel specific testing at the POC.