Background: Biomedical waste (BMW) refers to waste generated during the diagnosis, treatment, or immunization of humans or animals, posing risks to health and the environment if not managed properly. Proper knowledge, attitudes, and practices (KAP) among healthcare professionals are essential for effective BMW management. This study assessed the KAP of healthcare workers regarding BMW in a tertiary care teaching hospital. Methods: A cross-sectional study was conducted among 149 healthcare professionals at a private medical college from April 10 to May 10, 2023. Participants were selected using simple random sampling, including consultants, postgraduates, interns, nursing staff, laboratory technicians, and housekeeping staff. Data were collected via a semi-structured questionnaire covering socio-demographics and BMW-related KAP. Analysis was done using SPSS 28, with frequencies and percentages calculated for categorical variables. Results: Of the 149 participants, 98.7% had heard about BMW management, and 97.3% were aware of proper handling practices. While 98% adhered to color-coded waste segregation, only 21.5% used hub cutters for needle disposal, raising concerns about needle-stick injuries. Despite 71.8% having received BMW training, gaps in practical application remained. Attitudes were positive, with 94.6% recognizing BMW management's importance, and 83.9% willing to attend further training. Additionally, 83.2% of participants were vaccinated for Hepatitis B. Conclusion: While healthcare professionals demonstrated high awareness and positive attitudes towards BMW management, low compliance with needle disposal practiceshighlights the needfor regular training and stricter adherence to protocols. Improved practices can reduce injury risks and ensure a safer healthcare environment. Keywords: Biomedical waste management, waste segregation, compliance