Attachment of biomolecules on gold, silicon or glass surfaces has direct implications for the development of novel biosensors in the context of nanoscale detection of pathogens and other metabolites related to issues of human health. In this critical review, we have highlighted the current developments in various techniques of immobilization of biomolecules, specifically biological macromolecules on surfaces through the modification of a functional self-assembled monolayer. The utility of such immobilized biomolecules in the area of biosensing in nanoscale has been surveyed. Merits and demerits of some of the methods with reference to sensitivity of detection and practical use have been discussed (221 references).