Challenges associated with nonspecific adsorption of proteins on sensor surfaces have steered the development of novel antifouling materials and strategies. Inspired by human synovial fluid composition and structure, we designed synergistic antifouling coatings with mixtures of hyaluronic acid (HA) and a zwitterionic bottlebrush polymer (BB). Using a fast and convenient online surface modification method, the polymers were immobilized on the Au surface, significantly increasing its hydrophilicity. Using surface plasmon resonance (SPR), a 10:1 ratio of HA to BB was found optimal to provide the best antifouling performance. Bovine serum albumin (BSA) adsorption on HA-BB coated surfaces was 0.2 ng/cm2, which was 60 times lower than BB or HA alone and 25 times lower than the commonly accepted ultralow adsorption limit (<5 ng/cm2), demonstrating the synergistic effect of HA and BB against nonspecific protein adsorption. This was found to be independent of BSA concentration up to physiological concentrations. Furthermore, the antifouling performance of HA-BB coated surfaces was tested against milk and serum, showing almost 92% lower protein adsorption than that on bare surfaces, suggesting the potential efficacy of this antifouling coating in real life settings.