Label-Free Immunoassay Using Droplet-Based Brewster's Angle Straddle Interferometry

We report the detection of antigen capture by immobilized antibodies using a simple, label-free version of monochromatic reflective interferometry. The technique is implemented on silicon with its native oxide and relies on choosing an incident angle between the Brewster angles for the air/oxide and oxide/silicon interfaces. We demonstrate sensitivity to anti-human and anti-rabbit immunoglobulin (anti-IgG) concentrations less than 100 nM using only 10 nL droplets of the analyte. We have introduced a protocol using a model sugar to reduce nonspecific binding and have been able to detect anti-IgG even in the presence of 100-fold larger concentrations of bovine serum albumin. The limit of detection is not yet associated with the optical method but is imposed by nonspecific binding. Evaluated in terms of pg/mm², our sensors are comparable in sensitivity to surface plasmon resonance (SPR) but are advantaged with respect to SPR in the tolerance of the optical components and alignment, the low material usage, and the ability to exploit multiplex detection without modification. The simplicity and convenience of the method are promising for eventual application to portable diagnostic applications.