European protein production platforms
The EU-funded 'Infrastructure for protein production platforms' (PCUBE)(opens in new window) project worked on developing, improving and standardising protein production, screening and crystallisation technologies. PCUBE members were highly successful in their endeavours. To realise high-throughput production, they developed robot-friendly protocols for automation. They also optimised and standardised molecular biology methods for protein selection and 3D characterisation as well as DNA purification to improve protein quality. Other areas of focus included optimising light microscopy techniques and establishing efficient protein expression systems in prokaryotic, insect and mammalian cells. TNA was granted to 384 scientists and their projects from 30 different European countries. In parallel, 10 training events were offered on protein production, purification, expression, characterisation and crystallisation methods as well as protein engineering technologies. Proteins' production in high quality and quantity is now possible through systems such as the MultiBac uses a baculovirus to reprogramme insect cell cultures and is harmless to humans. They optimised it to produce large numbers of desired multi-protein complexes. Many European scientists accessed this patented technology through PCUBE TNA to produce proteins-of-interest. For instance, SweetBac was engineered that has untapped potential with regard to antibody production. PCUBE funding helped realise the automation of a protein complex screening method that led to the development of the CoESPRIT technology. This was utilised by TNA users to further optimise high-throughput cloning. Automation for the efficient selection and production of designed ankyrin repeat proteins (DARPins) was also funded by PCUBE. Several binders were developed for specific protein targets using DARPin technology and distributed to TNA users for research. The project also enabled the in vivo monitoring of live cells and ongoing protein interactions through techniques such as fluorescence cross-correlation spectroscopy and spectral position determination microscopy. Low-cost high-throughput technologies for protein crystallisation and in situ X-ray diffraction were developed that are robust and accurate. PCUBE has successfully brought about technological innovation by setting benchmarks in structural biology and TNA to high precision instrumentation via joint research activities. Besides basic research, applications of PCUBE technologies include food security, environment protection, biomedicine, green energy and biosensing.
