Holographic surface relief diffraction gratings made of hydrogels for direct label-free biosensing of IgGs

This work describes the development of a label-free (LF) biosensing platform for the direct detection of targets based on diffractive structures fabricated with acrylamide-based hydrogels biofunctionalized with proteins and antibodies. Hydrogels containing Bovine Serum Albumin protein (BSA) with different crosslinking degrees were synthesized and characterized to find the optimal conditions for the suitable fabrication of surface relief gratings (SRGs). The bioavailability of BSA-functionalized hydrogels for the specific recognition of anti-BSA antibodies was verified by fluorescence detection. After the hydrogel-based SRG fabrication, diffraction efficiency measures at two different laser wavelengths were used for the direct LF detection of anti-BSA antibodies. The limit of detection in the sub mg L^-1 range was read. Additionally, SRGs were prepared with hydrogels biofunctionalized with anti-rabbit antibodies for the direct detection of IgGs from rabbit serum, obtaining similar analytical performance without the necessity of labeling or applying amplification strategies.