ObjectiveTo assess the association between work location and movement behaviours (physical activity [PA], screen time, sleep) and adherence to the Canadian 24-Hour Movement Guidelines (24-H Guidelines) among Canadian workers during the COVID-19 pandemic. MethodsUsing cross-sectional data from the 2021 Canadian Community Health Survey (n = 10,913 working adults 18–75 years), primary work location was categorized as: worked outside the home at a fixed location (fixed workplace), worked at home (telework), and worked outside the home at no fixed location (non-fixed workplace). Recreational, transportation and occupational/household PA, as well as leisure screen time and sleep duration were self-reported. Logistic regression assessed associations between work location and adherence to movement behaviour recommendations, adjusting for covariates. ResultsCompared to a fixed workplace, those teleworking reported more recreational PA (21.1 vs 17.0 min/day, p < 0.0001) and sleep (7.2 vs 7.1 h/night, p = 0.026) and were more likely to meet sleep duration recommendations (adjusted odds ratio [aOR] = 1.28, 95% CI: 1.08–1.51) and the 24-H Guidelines (aOR = 1.25, 95% CI: 1.04–1.51). Compared to fixed workplaces, those at non-fixed workplaces reported more occupational PA (62.7 vs 32.8 min/day, p < 0.0001) and less leisure screen time (2.5 vs 2.7 h/day, p = 0.021), and were more likely to meet the PA recommendation (aOR = 1.46, 95% CI: 1.15–1.85) and the 24-H Guidelines (aOR = 1.38, 95% CI: 1.09–1.75). ConclusionsResults suggest that adherence to the 24-H Guidelines varies by work location, and work location should be considered when developing strategies to promote healthy movement behaviours. Future studies could explore hybrid work arrangements, and longitudinal study designs.