The project will develop the concept of employing Novozymes engineered proteins coupled to polymers, to form conjugates that will self-assemble into supramolecular micellar or vesicular architectures. These materials will contain amphiphilic block co-polymers that self-assemble under certain conditions, but which can disassemble under other conditions, for example by changes in temperature or pH. As a consequence it should be possible to encapsulate drug/biotherapeutic compounds in the interior of the self-assembled conjugates, but also to trigger the release of the therapeutic when conditions disfavour self-assembly. The proposal aims to combine the advantageous properties of both engineered proteins (serum stability, target specificity, biodegradability) with synthetic or modified natural polymers (tailored physical properties, tuneable functionalities) to generate a versatile new class of bioactive materials. The polymer components, will be based in the first instance on the well-known Pluronic series of amphiphilic co-polymers, to enable temperature-triggered assembly/disassembly, or hyaluronic acid, to enable ionic and pH-triggered assembly/disassembly. The other, biomolecular, component of the conjugate will consist of molecules available within Novozyme's product portfolio (including recombinant albumin (recombumin), albufuse, hyaluronic acid and recombinant transferrin (including modified versions for subsequent conjugation)). The latter components will be selected dependent on the functionality desired within the self-assembled complex. For example, transferrin could be utilized for improved cellular targeting and uptake, whereas albufuse would be employed to promote serum stability and half-life. Combinations of these conjugates with other protein-polymer hybrids and/or block co-polymers will thus enable a family of self-assembling materials to be developed, that will display varying degrees of transferrin, or transferrin-type functionality, as well as controllable protein activity dependent on self-assembly state. Most importantly, the controllable assembly/disassembly of the conjugates will allow the encapsulation and release of therapeutic agents/actives in the micellar core or vesicular interiors of the conjugates. As such, the conjugates represent a new materials platform that may be applicable to many areas of therapeutic, flavour and fragrance release. The student will prepare polymer-protein conjugates using techniques well-established in the School of Pharmacy (Chem. Commun. 2008, 4433 - 4435, Int. J. Pharmaceutics 2007, 340, 20-28, J. Am. Chem. Soc. 2004, 126, 13208-13209). A key component of their studies will also be the biophysical characterization of the developed conjugates and in particular their self-assembly/disassembly behaviours. This will be achieved to the nanometre scale using the suite of instruments available within the School (including atomic force microscopy (AFM), Quartz-Crystal Microbalance (QCM-D), CPS-Disc Centrifuge and light light scattering (DLS)). Importantly these tools will also be employed to probe interactions with model cellular membranes (building on recently published studies (Molecular Biosystems 2008, 4, 741-745)), and also cellular interaction and uptake (using scanning ion condictance microscopy (SICM) and confocal microscopy). Encapsulation of actives (for example doxorubicin, nucleic acids) and their release from self-assembled bioconjugates will be evaluated by fluorescence and UV-Vis spectroscopies and/or gel-shift assays for biotherapeutics. To assess the efficacy of materials (e.g. drug encapsulation and release within cell models), the student will work closely with researchers within on-going drug-delivery projects within the School. Importantly the developed conjugates will be evaluated in comparison with conventional drug delivery systems (liposomal doxorubicin, Lipofectamine) as well as protein-nanoaggregate carriers (e.g. albumin-paclitaxel (Abraxane))

Steering nanoparticle transport in human cells - why is this important? Viruses seem to travel effortlessly into tissues and cells, being transported selectively in the body to reach the sites where they can cause most harm. They do this by breaching cellular barriers such as the outer or plasma membrane of cells and use human cellular machinery to make copies of themselves. Drug molecules on the other hand spread non-selectively throughout the body thus reducing effects where they are most needed, and causing adverse side-effects where they are not wanted. One reason for this is that current synthetic carriers for drugs and diagnostic agents, unlike viruses, are unable to effectively cross biological barriers and then reach specific sites inside cells. An artificial particle that could transport through tissue, in a manner analogous to a virus and then into defined cell locations but without causing disease, would therefore revolutionise healthcare applications. This would be particularly important for the early stage diagnostics and therapeutics needed in developing nations and for ageing populations. What do novel polymer-coated gold nanoparticles have to offer? These materials are an optimal test platform for proof-of-concept studies described in this application. Firstly, gold particles can be tracked in cells using a number of microscopic approaches including the newly developed highly sensitive four-wave mixing imaging system available to this team. They can be coated with a variety of polymeric materials that will help to guide them into cells and into specific cell locations. We have previously shown that polymers which are capped with functional 'keys' to enter natural cell portals can have their entry switched on and off by small increases in temperature which cause them to change their conformations. We also have shown that we can generate these temperature increases at gold nanoparticles inside cells through laser pulses but without damaging the cells. By attaching the temperature-responsive polymers to gold, it should be possible to use laser pulses to switch the functional keys on and off, and in this way guide particles to reach defined cellular locations. This will help to unravel the mechanisms by which materials travel in cells, thus enabling us to guide diagnostics and therapeutics to where they are required. Impact. A major hurdle to effective therapy against major disease burdens such as cancer, coronary heart disease and neurodegeneration is our inability to direct therapeutic molecules such as genes and proteins to specific tissue and defined compartments inside cells. This is a major objective of this application and progress here could have widespread implications for academia, industry and the society that they serve. One could envisage a commercial application in which a combined imaging and guiding instrument (e,g, ultrasonic probes and imaging) is used with a set of nanoparticles with functionality for specific disease markers, with a potential for truly selective personalised therapies. Better diagnostics are also needed that allow earlier detection of disease and thus better healthcare outcomes. Successful completion of this work could allow development of an imaging/detection platform where specific markers of disease could be detected through their interaction with selective receptors on gold nanoparticles guided to intracellular sites by the local laser-heating method. When it is considered that 1 in 3 individuals in the EU will be affected directly or indirectly by cancer by 2010, it is clear that earlier detection and intervention will bring marked benefits to patients, carers and society as a whole. Longer-term development could generate impact through a new biomedical technology i.e. laser-guided therapeutics wherein local heating by focused ultrasound guides biodegradable responsive nanoparticles in humans.

Biopharmaceutical manufacturing continues to evolve with an increased emphasis on underpinning science and engineering. Effective deployment of contemporary knowledge in science and engineering throughout the product life cycle will facilitate manufacturing efficiencies and regulatory adherence for biopharmaceuticals. Fundamental to this paradigm shift has been the drive to adopt an integrated systems approach based on science and engineering principles for assessing and mitigating risks related to poor product and process quality. Changes have been enabled as a consequence of the regulatory authorities introducing a new risk-based pharmaceutical quality assurance system. The traditional approach to manufacture has been to accommodate product variability into the specifications and fix operational strategies to ensure repeatability. Developments in measurement technology have invited changes in operational strategy. This revised approach is based on the application of Quality by Design (QbD), underpinned by process analytical technology (PAT) to yield products of tighter quality and more assured safety. QbD is defined as the means by which product and process performance characteristics are scientifically designed to meet specific objectives. Practical improvements therefore demand a knowledge base of science and engineering understanding to identify the interrelationship between variables and integrate the learning into different manufacturing scenarios. The focus of the Centre is to address the challenges emerging from this paradigm shift and to train a new generation of students with competencies in all stages of commercial biopharmaceutical process development. Critical to this is to ensure they have the skills to work at the discipline interfaces in the areas of biosystem development, upscaled upstream process engineering, and the engineering and development of downstream processing. The training will be formulated around three elements that form the backbone of achieving an enhanced understanding of the process. The three elements are (i) Measurement, Data and Knowledge Management, (ii) Enhance Available Knowledge and (iii) Use Knowledge More Effectively. The power of the approach being adopted is that it is equally applicable to established bioprocesses based on microbial and animal cell culture, as well as emerging areas including stem cells, marine biotechnology and bio-nanotechnology. The rationale for proposing a Centre in this area is to address a well recognised problem, a lack of appropriately trained personnel, who will deliver the next generation of biopharmaceutical development. These issues have been clearly articulated in a series of reports. SEMTA reported that over a quarter of bioscience companies do not have sufficient science skills. 39% of bioscience/pharmaceutical companies have long-term vacancies; with 22% having skill shortages in the science arena (five times that for other sectors). Lord Sainsbury, concerned at the rapidly changing nature of the bioscience business, set up the BIGT and commissioned Bioscience 2015. One of the strong messages raised was the serious shortfall in trained staff. Furthermore a quantitative assessment of the increase needed of trained people entering the sector was made by bioProcessUK. They estimated an increase of 100 trained personnel was required on top of the current 150 doctoral level candidates graduating per year. It is not simply a matter of increasing the number of trained persons. The Centre will also address the limitations of the current UG training of engineers, chemists and biologists which does not prepare them for the challenge of working in process development distinguished by disciplinary interfaces. The proposed programme will address a strategic shortfall and produce a new generation of graduates with the appropriate inter-disciplinary skills to drive both the research agenda and knowledge transfer of underlying concepts into industry.

In the 1980s it began to be possible to produce potentially unlimited quantities of human proteins by placing the gene defining them in a simple organism such as yeast. From this grew a new kind of medicine capable of treating conditions such as severe arthritis, haemophilia, growth deficiency, and some cancers that previously had no satisfactory treatments. As well as having great clinical value the resulting technology has become the basis of a new and fastest growing part of the pharmaceutical industry, described as biopharmaceuticals. Because the molecules involved are proteins, they are orders of magnitude larger and more complex than conventional drugs such as aspirin and their processing is much more demanding. They are also so complex that they cannot in general be characterised with precision except in relation to the methods by which they are made. That means the capacity to precisely define such processes is critical to clinical safety and commercial success. Full scale trials of the processes are so costly they can only be conducted once clinical promise is established but, given the number of factors governing processing of even first generation products, there have often been hold-ups so extensive as to delay availability to patients. UCL has pioneered micro scale methods that are sufficiently good at predicting efficient conditions for large scale performance that far fewer and better focussed large scale trials suffice. That resolves part of the problem but an even greater challenge is now emerging. The early biopharmaceuticals were in general the easiest ones to produce. The final scales were also relatively modest. Now, the next generation of biopharmaceuticals are more complex materials and with rising demand the scales are far larger so that processes push the boundaries of the possible. The combined complexity of the product and the process with so many variables to consider means that the managers need better systematic means of supporting their decisions. Already the cost of developing a single biopharmaceutical can exceed 0.7 billion and take 10 years. With more advanced biopharmaceuticals these figures tend to rise and yet the world's governments are facing a healthcare cost crisis with more older people. They therefore exert pressure on companies to reduce prices. Because the public wishes to have medicines that do not pose risks, regulations become ever more stringent so they are a major factor in defining the bioprocess. This also adds to the need for managers to have sector-specific decisional-support aids well grounded in the detailed engineering of the processes. Finally, it is now possible to apply molecular engineering to proteins and vaccines to enhance their therapeutic properties but this can also cause serious bioprocessing problems. The research vision developed with detailed input from UK industry experts will apply these methods as the foundation for another step change whereby much faster and lower cost information can be gathered and integrated with advanced decisional techniques to give managers a better foundation on which to base their policies. The academic team from leading UK universities provides the necessary continuum of skills needed to assess the ease of manufacture of novel drugs, the costs of processing and of delivery to patients. We will work with companies to test the outcomes to ensure they are well proven prior to use on new biopharmaceuticals. This will cut costs so that all the patients who might benefit can receive them and at the earliest possible date achieved within the severely restricted budgets now available to the NHS.