Certain prophylactic measures including mass vaccination are being adapted to avoid severe economic losses caused by the endemics of Newcastle disease. Live vaccines in liquid phase cannot be preserved for longer time due to different physical and chemical degradation occurring in the aqueous solution but this degradation can be significantly reduced by reducing the water contents of vaccines through the process of lyophilization. Present study was designed to develop and optimize the lyophilization process for the preparation of lentogenic (LaSota) vaccine by comparing five different protocols devised for this purpose (28:00 hours for Protocol A, 25:20 hours for Protocol B, 21:20 hours for Protocol C, 18:15 hours for Protocol D and 15:15 hours for Protocol E). Variable time durations for freezing, primary and secondary drying were given in each protocol. The resultant vaccine from each protocol was assessed for its physical appearance, haemaggltination titers, safety in laboratory as well as in host animals, sterility using different biological media, residual moisture content and vaccine potency. Vaccine obtained from Protocol A (28:00 hours) was found satisfactory in terms of all aforementioned parameters. The findings of this study can help the vaccine manufacturers for the production of lyophilized live LaSota vaccine.