The adsorption of non-specific biomolecules onto solid supports creates problems of sample loss and reduction in the sensitivity and performance of sensors used for in vitro diagnostics in food and health monitoring. In an effort to develop bioactive surfaces that resist non-specific biomolecules adsorption, the surfaces are generally functionalized with poly(ethylene glycol) (PEG) or oligo(ethylene glycol) which effectively drives away the non-specific biomolecules and enhances their bioactivity. In this work we have reviewed selective strategies for surface functionalization with hydrophilic polymers so that bioactive molecules can be preferentially attached to these surfaces to develop protein-specific and protein-resistant antifouling bioactive surfaces. Adhesion strategies responsible for antifouling over polymeric substrates and their effects on improved, efficient and reproducible electrophoretic separation of biomolecules over solid surfaces have been reviewed. © 2015 Scrivener Publishing LLC.