Biosensors are devices that detect the presence of an analyte with a bioreceptor and transduce a measurable signal. They are widely used in medical devices such as glucose monitors, rapid antigen tests and pregnancy tests, and for the detection of pollutants and pathogens in food, water, and the environment. Protein biosensors have been developed to target a wide variety of molecules by using antibodies or other proteins as bioreceptors and fluorescent proteins or enzymes for signal transduction. Many protein biosensors consist of two protein components that form a complex upon ligand detection. Although 2-component protein biosensors are simpler to develop, they exhibit several disadvantages, including their limited utility at low concentrations and when immobilised on a surface. In this study, I have demonstrated that attaching two biosensor components with a molecular linker to convert them into a 1-component biosensor is an effective and generalisable method for increasing the sensitivity of 2-component biosensors. I first showed with numerical methods that 1-component biosensors are more sensitive than 2-component biosensors and have an increased dynamic range, and that the magnitude of the change in these properties depends on the properties of the linker. I then experimentally validated the model by assembling a fully integrated 1-component biosensor on a tunable DNA scaffold. The biosensor consists of a beta-lactamase enzyme that is unfolded by the insertion of a calmodulin protein, but refolds upon the binding of a ligand to receptors that are fused to the enzyme and a calmodulin-binding peptide. The proteins are attached to DNA through a SpyTag peptide-DNA conjugate. I demonstrated the synthesis, assembly, and function of the 1-component biosensor and compared its activity and sensitivity to that of the 2-component system. These results demonstrated that 1-component biosensors do indeed have an enhanced signal that is dependent on the characteristics of the linker. By exploring the differences between 1- and 2-component biosensors, this study has established a foundation for the optimisation of linker domains in 1-component biosensors, facilitating the conversion of two-component biosensors into 1-component systems to enhance their sensitivity. I anticipate that this work will inform the design of ultra-sensitive 1-component biosensors, furthering the accurate detection and quantification of molecules that are critical for environmental and human health.