With increasing emphasis on green chemistry, biomass-based materials have attracted increased attention regarding the development of highly efficient functional materials. Herein, a new pore-rich cellulose nanofibril aerogel is utilized as a substrate to integrate highly conductive polypyrrole and active nanoflower-like nickel-cobalt layered double hydroxide through in situ chemical polymerization and electrodeposition. This ternary composite can act as an effective self-supported electrode for the electrocatalytic oxidation of glucose. With the synergistic effect of three heterogeneous components, the electrode achieves outstanding glucose sensing performance, including a high sensitivity (851.4 µA·mmol⁻¹·L·cm⁻²), a short response time (2.2 s), a wide linear range (two stages: 0.001−8.145 and 8.145−35.500 mmol·L⁻¹), strong immunity to interference, outstanding intraelectrode and interelectrode reproducibility, a favorable toxicity resistance (Cl^‒), and a good long-term stability (maintaining 86.0% of the original value after 30 d). These data are superior to those of some traditional glucose sensors using nonbiomass substrates. When determining the blood glucose level of a human serum, this electrode realizes a high recovery rate of 97.07%–98.89%, validating the potential for high-performance blood glucose sensing.