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Surface assays such as ELISA are pervasive in clinics and research andpredominantly standardized in microtiter plates (MTP). MTPs provide manyadvantages but are often detrimental to surface assay efficiency due toinherent mass transport limitations. Microscale flows can overcome these andlargely improve assay kinetics. However, the disruptive nature of microfluidicswith existing labware and protocols has narrowed its transformative potential.We present WellProbe, a novel microfluidic concept compatible with MTPs. Withit, we show that immunoassays become more sensitive at low concentrations (upto 9-fold signal improvement in 12-fold less time), richer in information with3-4 different kinetic conditions, and can be used to estimate kineticparameters, minimize washing steps and non-specific binding, and identifycompromised results. We further multiplex single-well assays combiningWellProbes kinetic regions with tailored microarrays. Finally, we demonstrateour system in a context of immunoglobulin subclass evaluation, increasinglyregarded as clinically relevant.

Advection-enhanced kinetics in microtiter plates for improved surface assay quantitation and multiplexing capabilities