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Rapid, reliable and low cost techniques to fabricate biosensors is a hot topic nowadays. Here, we present a BIO-grating fabricated by means of local, selective denaturing of molecules using UV radiation. A phase-mask is used to generate an interferometric pattern of 1420 nm pitch that, when illuminating a biolayer of BSA molecules lead to its periodic deactivation. After the biorecognition of the specific antibody, aBSA, a BIO-grating is generated due to the height difference between the protein, and the complex protein + antibody. We present the optimization of the fabrication of the BIO-gratings and their AFM characterization. Also, the biosensor performance in terms of limit of detection and limit of quantification will be presented.