The capacity of exclusive two-wheeler lanes in India has become a critical topic of interest in recent years as these lanes are gaining popularity as a solution to traffic congestion and accidents. The purpose of this study is to study the capacity of exclusive two-wheeler lanes in Nagpur and identify those factors that affect their capacity. The study employs a combination of field observations, traffic counts, and surveys to determine the capacity of exclusive two-wheeler lanes. It also analyses the factors that affect capacity, such as lane width, and traffic flow characteristics. This study aimed to investigate the capacity of exclusive two-wheeler lanes with varying widths of 2, 2.5, and 3 m in Nagpur city and find that the capacity of exclusive two-wheeler lanes is influenced by various factors, including the width of the lane, the speed of two-wheelers, and the presence of obstacles on the lane. The Kinovea, K-lite Codec, and Excel software were used for data extraction and VISSIM software for microscopic simulation to analyse the capacity. Additionally, the speed limits and design speed were calculated using the cumulative frequency distribution method to find the 15th, 85th, and 98th percentile speeds. The study also identifies the need for proper signage and markings to enhance the capacity of these lanes.