This paper includes sensitivity assessment of label-free biosensors using hetero dielectric Junctionless-TFET (HD-JL-TFET) thorough TCAD simulator. The fundamental structure, operation and design of a Junctionless-TFET (HD-JL-TFET) as biosensor are investigated in this paper. For the purpose of detecting the biomolecule, a nano-gap is added close to the source end between the gate and channel. To test the sensing potential, we adjusted the charge density and material dielectric constant (K) by comprehensive calibrated device simulation. For several biomolecules, the device's sensitivity was examined as surface potential, electron tunnelling rate, and conduction-valence band edge fluctuation. Additionally, the Id vs VGS features, the sensitivity to the drain current, and the ION/IOFF fluctuation are also examined. By contrasting neutral or charged biomolecules using various dielectric constants, the sensitivity characteristics of positive, negative, and neutral biomolecules are examined. The development of biosensors, which enable the rapid and precise detection of multiple biomolecules, has revolutionized the field of bioanalysis.