Biotechnology is a key area of industrial interest and the importance of effective knowledge management for rapid bioprocess development, optimisation and operation is widely recognised as an important driver of biomanufacturing excellence. The Britest suite of tools and methodologies, designed to highlight knowledge gaps within chemical and physical processes, is explored as an approach to bioprocess knowledge acquisition and management. These tools can help identify where optimisation may be most beneficial, and also increase understanding of the process as a whole across a range of disciplines. This research identifies areas where Britest tools are not directly transferable into biotechnological applications, and formulates a whole bioprocess development methodology. The Britest tools have been considered using SuperPro Designer in relation to production of insulin using E. coli. Some of the existing Britest tools have been found to be directly applicable to biological processes, although adaptations were required in some cases, to account for differences between chemical and biological processing. A gap was identified relating to considering the process as a whole, and so a new tool (the Reaction/Reagent/Transformation Tracker, R2T2) was developed to address this. The Britest tools show promise within the context of a bioprocess, although further work is required to fully realise their potential in this exciting field. It is anticipated that the tools can be applied to aid in the identification of Critical Quality Attributes (CQAs) and Critical Process Parameters (CPPs), constructing a robust design space, and facilitating the development of a Quality by Design (QbD) approach to bioprocessing.