Nanomaterials have revolutionized scientific research due to their exceptional physical and chemical capabilities. Carbon-based nanomaterials such as graphene and its derivates have excellent electrical, optical, thermal, physical, and chemical properties that have made them indispensable in several industries worldwide, including medicine, electronics, and energy. By incorporating carbon-based nanomaterials as nanofillers in electrospun nanofibers (ESNFs), smoother and highly conductive nanofibers can be achieved that possess a large surface area and porosity. This approach provides a superior alternative to traditional materials in the development of improved biosensors. Carbon-based ESNFs, among the most exciting new-generation materials, have many applications, including filtration, pharmaceuticals, biosensors, and membranes. The electrospinning technique is a highly efficient and cost-effective method for producing desired nanofibers compared to other methods. Various types of natural and synthetic organic polymers have been successfully utilized in solution electrospinning to produce nanofibers directly. To create diagnostics devices, various biomolecules like antibodies, enzymes, aptamers, ligands, and even cells can be bound to the surface of nanofibers. Electrospun nanofibers can serve as an immobilization matrix to create a biofunctional surface. Thus, biosensors with desired features can be produced in this way. This study comprehensively reviews biosensors that integrate nanodiamonds, fullerenes, carbon nanotubes, graphene oxide, and carbon dots into electrospun nanofibers.