Knowledge transfer is a key prerequisite for the success of setting up management systems in organisations, as they become one of the strategic resources that can ensure increased performance or significant growth. There are a number of strategies using multiple activities (patent sales, custom research, consulting, horizontal mobility, publishing activities, human resources management practices) that ensure effective knowledge transfer for achieving long-term competitive advantage. These initiatives are considered as an important aspect of knowledge management and sustainable business development. The article aims to identify and evaluate a group of key factors to ensure the transfer of knowledge in selected Czech organisations. Data were obtained through a questionnaire technique of data collection (n = 183) using descriptive and multidimensional statistics in combination with a focus group (n = 8). Four key factors have been identified to ensure knowledge transfer, namely internal factors, altruism, satisfaction, organisational identity.