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Background: Biomedical waste (BMW) management is essential for hospital infection control and environmental safety, and inadequate segregation, limited awareness of legal frameworks, and poor access to policy documents increase occupational hazards and the risk of nosocomial infections. Objectives: To assess knowledge and practices related to biomedical waste management and associated legal frameworks among doctors and nurses in a tertiary care hospital in Mysuru, India. Methods: A descriptive cross-sectional study was conducted among doctors and nurses at a 250-bed tertiary care teaching hospital in Mysuru, Karnataka. Data were collected using a structured, self-administered questionnaire assessing knowledge of BMW guidelines, access to policy documents, waste handling and segregation practices, and awareness of infection risks. Data were analysed using descriptive statistics and the Chi-square test. Results: Of the 150 questionnaires distributed, 128 were completed (response rate: 85%). Nurses constituted 74.2% of participants and doctors 25.8%. Awareness of the WHO manual on safe management of health-care waste was limited (45.3%), with no significant difference between professional groups (p = 0.40). Access to BMW policy documents differed significantly, with 90.5% of nurses reporting access compared to 15.2% of doctors (p < 0.001). Despite limited policy awareness, more than 90% of respondents reported appropriate waste segregation, correct use of colour-coded bins, and glove use. Awareness of infection risks associated with improper BMW management was high. Conclusion: Although reported biomedical waste management practices were satisfactory, gaps in awareness of and access to formal guidelines and legal frameworks persist, particularly among doctors. Targeted training and improved dissemination of policy documents are required to strengthen compliance.