Direct whole blood analysis by the antigen-antibody chemically-delayed dissociation from nanosensors arrays

A wide range of fields, starting from basic research in life sciences and up to medical applications, are highly interested in the investigation and detection of biomarkers in all their forms, including proteins. However, direct analytical detection of specific protein biomarkers from a physiological biosample is still extremely challenging due to the abundant variety and amount of its components. In this work, we apply the chemically-controlled antigen-dissociation detection approach on silicon nanowires-based field-effect transistor arrays, by creating a suitable 'chemical environment' which enabled the clear-cut splitting of the dissociation regime window into two sub-regimes, thus allowing the complete washing of the nonspecifically adsorbed salts and biomolecules, while significantly delaying the dissociation of specific surface-bounded antigen-antibody pairs. This was accomplished by the addition of the water-miscible organic reagent ethylene glycol, which radically alters the properties of the aqueous solvent, by means of dramatically reducing its interactions with the particular protein antigen, and thus allowing for the increase in the antigen-antibody interaction strength. This in turn, deeply reduces the solubility of the surface-bound protein molecules and increases their interaction with the specific receptor antibody units, which brings to a substantial delay in the antibody-antigen dissociation behavior. This phenomenon allows the clear-cut splitting of the dissociation regime window and the quantitative and accurate analysis of proteins in physiological samples. We demonstrated the direct and quantitative detection of protein biomarkers, down to concentrations in the fM range, from unprocessed whole blood minuscule samples of only a few microliters. This work is the first demonstration on the chemically-controlled dissociation kinetics of antibody-antigen pairs by the use of water-miscible organic solvent mixtures, and its application in the direct ultrasensitive detection of protein biomarkers from whole blood samples.