Development of bioassays has become the matter of intense research in the field of molecular diagnostic. Biodetection techniques have been drastically used in laboratory since the past 20 years and tend now to reach the in-vitro diagnostic industry. Most commercially available biosensing methods rely on immunoassays and use fluorescence as reading technique. However, the use of labeling methods such as fluorescence increases the cost of a single bioassay and may interfere with the biological functions of molecules. In this perspective, we have developed an optical label-free technique of microarray reading, which is an alternative to fluorescence. In this work, we use a label free biosensing method based on the diffraction of light by molecular gratings. Molecular gratings are employed as diffractive probe arrays for protein interaction analysis, as the diffraction efficiency changes in response to analyte binding. This Ph.D is supported by the French company Innopsys, which provides optical solutions for microarray reading and the Nanobiosystems group at the LAAS-CNRS. This work deals with the development of a detection platform for biomolecular interactions analysis, through the use of multiplexed biochips and the validation of an optical scanner. We present a special surface chemistry, based on blocking layers to reduce the non-specific protein adsorption and consequently decrease the limit of detection. Thanks to bi-functionalized biochips and this label free instrument, we have detected proteins interactions involving low molecular weight molecules.