Real-Time study of the adsorption and grafting process of biomolecules by means of bloch surface wave biosensors

A combined label-free and fluorescence surface optical technique was used to quantify the mass deposited in binary biomolecular coatings. These coatings were constituted by fibronectin (FN), to stimulate endothelialization, and phosphorylcholine (PRC), for its hemocompatibility, which are two properties of relevance for cardiovascular applications. One-dimensional photonic crystals sustaining a Bloch surface wave were used to characterize different FN/PRC coatings deposited by a combination of adsorption and grafting processes. In particular, the label-free results permitted to quantitatively assess the mass deposited in FN adsorbed (185 ng/cm2) and grafted (160 ng/cm2). PRC binding to grafted FN coatings was also quantified, showing a coverage as low as 10 and 12 ng/cm2for adsorbed and grafted PRC, respectively. Moreover, desorption of FN deposited by adsorption was detected and quantified upon the addition of PRC. The data obtained by the surface optical technique were complemented by water contact angle and X-ray photoelectron spectroscopy (XPS) analyses. The results were in accordance with those obtained previously by qualitative and semiquantitative techniques (XPS, time-of-flight secondary ion mass spectrometry) on several substrates (PTFE and stainless steel), confirming that grafted FN coatings show higher stability than those obtained by FN adsorption.