A reagent-free, low-cost and sensitive immunosensor has been fabricated using anti-apolipoprotein B (AAB) conjugated l-cysteine in situ capped cadmium sulfide quantum dots (CysCdS QDs) bound to nickel oxide nanorods (nNiO) for detection of low density lipoprotein (LDL) molecules in human serum samples. The structural and morphological properties of AAB conjugated CysCdS QDs and nNiO have been investigated using electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy and UV-visible techniques. In this immunosensor, the synthesized NiO nanorods act as mediators that allow the direct electron transfer due to their channeling effect resulting in a mediator-free biosensor. This mediator-free CysCdS-NiO based immunosensor shows improved characteristics such as a good sensitivity of 32.08 μA (mg dl-1)-1 cm-2 compared to that based on nNiO (1.42 μA (mg dl-1)-1 cm-2) alone for detection of lipid (LDL) molecules over a wide concentration range, 5-120 mg dl-1 (0.015-0.36 μM). The kinetic analysis yields an association constant (Ka) of 3.24 kM-1 s-1, indicating that the antibody conjugated CysCdS-NiO platform has a strong affinity towards lipid molecules. The excellent electron transport properties of the CysCdS-NiO nanocomposite in this immunosensor reveal that it provides an efficient platform for routine quantification of LDL molecules in real samples. © The Royal Society of Chemistry 2016.