Qkine combines proprietary animal-free biomanufacturing processes with protein engineering technology. Leveraging our expertise and manufacturing process development platform, we will develop a unique animal-origin-free, high-purity TGF beta family product portfolio to accelerate stem cell science, overcome specific drug discovery barriers, and solve fundamental reagent availability and scale challenges for high-growth fields including cell therapies, precision oncology, regenerative medicine, 3D organoid biology and cultivated meat. Additionally, we will address the currently intractable scale-up challenges associated with TGF beta proteins essential to establishing a credible TGF beta growth factor supply chain for the cultivated meat and cell therapy manufacturing sectors.

Organoids are 3-dimensional (3D) clusters of stem cells that come together and emulate the microenvironment within individual organs, whether that be liver, kidney, heart, gut or other specific organs. Essentially, they can be viewed as miniature, simplified organs. They typically range in size from a few micrometers to five millimeters and there are potentially as many different organoids as there are different tissues and organs in the body. Organoids can also be grown that mimic diseased such as cancer and brain disorders. Such a diverse range of organoids can be formed by controlling the differentiation of the specific stem cell used, which can be influenced by the cells receiving instructive signals from proteins called growth factors, the 3D extracellular matrix (ECM) and its components, and chemical moieties in the medium the organoids grow in. Organoids hold extraordinary promise: they are a truly disruptive technology capable of completely transforming our understanding of basic biology and revolutionising the drug discovery process, and its reliance on animal models. That said, methods used for growing organoids in the laboratory need to be improved and standardised to allow wider adoption, particularly in drug discovery, and to make organoid culture easier and more reproducible. The two elements that need particular focus are the bioactive protein growth factors and the 3D gel matrix in which organoids grow. The matrix, a complex mixture of structural proteins and growth factors, is currently extracted from mouse tumours so there is a pressing need to make synthetic alternatives for reproducibility and to reduce use of animal-products. In our previous UKRI-funded project we developed optimised, highly defined forms of growth factors for organoid culture. Large multinational companies are now interested in bulk purchase of these and we need to undertake further R&D to scale manufacture, remove the use of beta-lactam antibiotics (which are not compatible with pre-clinical applications of organoids) and introduce a quality system ISO13485. In addition, there is one growth factor, called Wnt3a, which is crucial for the growth of many types of organoids and is not easily sourced. The few commercially available have variable bioactivity, are exceptionally expensive (<£17,000 /milligram) and are not made in a full animal-product-free process. We made a promising start on the R&D needed to solve this problem in the last project and wish to continue this work, using protein engineering technologies to make a highly bioactive, totally animal-free and cost-effective Wnt protein.

Organoids are 3-dimensional (3D) clusters of stem cells that come together and emulate the microenvironment within individual organs, whether that be liver, kidney, heart, gut or other specific organs. Essentially, they can be viewed as miniature, simplified organs. They typically range in size from a few micrometers to five millimeters and there are potentially as many different organoids as there are different tissues and organs in the body. Organoids can also be grown that mimic diseased such as cancer and brain disorders. Such a diverse range of organoids can form by controlling the differentiation of the specific stem cell used, which can be influenced by the cells receiving instructive signals from the 3D extracellular matrix (ECM) and its components, such as bioactive proteins, and the medium the organoids grow in. Organoids hold extraordinary promise: they are a truly disruptive technology capable of completely transforming our understanding of basic biology and also revolutionising the drug discovery process, and its reliance on animal models. That said, growing organoids in the laboratory still requires the use of animal-derived components; in particular, the 3D gel matrix in which organoids grow, which is made from mouse tumours. There are very few suppliers of this matrix and because it is made from animals, each batch is slightly different, it is also unusual in that it is liquid at 4oC but sets to a gel at room temperature, this makes it very difficult for scientists to use and not compatible with robotic systems needed in high throughput drug discovery. Thus, to fully develop the potential of organoids in their capacity to reduce the need for use of animals in research, it is essential that a replacement for this type of 3D matrix is obtained, which is not derived from animals. The aim of this project is to develop a new, fully synthetic (non-animal derived) 3D gel matrix which is optimised for the growth of organoids and can be used in the future for industrial-scale organoid production to drive forwards biomedical research, drug discovery and development of new therapeutics. This will be achieved by combining the proprietary synthetic matrix from Manchester BIOGEL with the optimised bioactive cell signalling growth factor proteins from Qkine to create a wholly synthetic hydrogel that recreates the ideal growth environment for the organoids. Cellesce, a specialist organoid company will help tailor the synthetic hydrogels for different organoid types and downstream applications to maximise the impact on science and the commercial potential of the combined technology.

One of the most exciting developments in biological research is the development of miniature three-dimensional structures that mimic mammalian organs such as gut, pancreas, lung and, in the case of cancer, tumours. These so-called organoids allow us to study the structure, development and function of these organs in highly controlled environment and most importantly, without the need to use animals. Organoids are likely to revolutionise drug development by providing a platform in which to test the efficacy and possible toxicity of new drugs and they facilitate the development and modelling of individualised therapies. One of the critical factors in growing organoids is the use of a precise cocktail of specific proteins, growth factors and cytokines, to mimic the micro-environment in the body. These proteins are hard to make, expensive and their use is often limited by variable quality and commercial availability. We have expertise in developing variant forms of these proteins using structure-guided protein engineering to optimise the biological properties, production process and quality of these growth factors. Here, we will use our expertise to improve those growth factors critical for organoid culture. We will ensure the quality of our products by defining strict criteria for their evaluation and ensure they are formulated in the most effective way for routine use. The results of this project will establish UK-based manufacturing of critical components for the emerging field of precision medicine.