This project will look at a novel method of treating sepsis, one of the world’s leading causes of death. The method uses magnetic nanoparticles (MNPs) that bind to specific entities in blood, combined with MediSieve's patented magnetic blood filter. By infusing the MNPs into an extra-corporeal blood loop containing the filter, MediSieve can remove both the pathogens that cause sepsis (bacteria) and the endotoxins that are responsible for the dysregulated immune response which is characteristic of sepsis. Importantly the method would work even in antibiotic resistant cases. Annually, there are 1M cases in the US and 900K in Europe, with 135K deaths/year in the EU. Mortality rates are 29-50% with a cost of €25K per patient. With the aging of global populations sepsis incidence is increasing rapidly (8-13% annually in the decade to 2013). The challenge is that both the pathogens and the immune response contribute to the disease. The destruction of bacteria by the immune system or antibiotics creates large quantities of endotoxins, which in turn aggravate the immune response and cause a cascade towards septic shock. MNPs that bind specifically to gram-negative bacteria and endotoxins enable the two targets to be rapidly and safely removed from circulation using MediSieve’s magnetic filter. The method provides the unique capability to specifically target both root causes of sepsis, reduce both the mortality and the economic costs of sepsis. The objectives of this Phase 1 study are to validate the technical and commercial feasibility of the proposed treatment method. The latter includes detailed market research, business planning, healthcare economics modelling and a detailed investment proposal with timelines and return on investment projections. The technical assessment will include in vitro lab studies validating that the MNPs efficiently bind to both targets and captured by the magnetic filter. A Phase 2 project would include pre-clinical and clinical studies.

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at https://www.ukri.org/apply-for-funding/how-we-fund-studentships/. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.

Most licensed monoclonal antibodies (mAbs) contain a consensus N-linked glycosylation site on their heavy chains. The oligosaccharides attached to this site greatly influence the efficacy of mAbs as therapeutics either by reducing their serum half-life or by directly affecting the mechanisms they trigger in vivo. It has been widely reported that cell culture conditions, such as carbon source type and availability, dissolved oxygen concentration, ammonia concentration, medium pH and culture mode, affect protein glycosylation, thus having great impact on end product quality (1-4). Recently, the US FDA and the European Medicines Agency have proposed the implementation of the Quality by Design (QbD) paradigm to the manufacture of biopharmaceuticals. Its implementation requires the use of all available knowledge of a given product for the design, optimization and control of the manufacturing process. The goal is to ensure that quality is built into the product at every stage of the manufacturing process. It is proposed that detailed mathematical models may play a critical role in the design, control and optimization of biopharmaceutical manufacturing processes under the QbD scope. To our knowledge, there are currently no mathematical models that relate product quality in terms of glycosylation with cell culture conditions. A model with this capability would be useful for bioprocess design and control, culture media formulation and hypothesis testing for genetic engineering strategies. We propose the development of a novel mathematical tool that links bioprocess conditions to protein glycosylation. The tool will encompass four components: transport of nutrients into the cell, cytosolic synthesis of nucleotide sugar donors (NSDs), transport of NSDs from the cytosol to the Golgi apparatus, and, finally, their addition onto the protein of interest. Lack of carbon availability directly affects the intracellular availability of NSDs, without which the glycan metabolism in the Golgi cannot continue. Other studies have shown that high ammonia concentrations (4) and extremes of pH (5) lead to poor glycoprofiles. Process conditions therefore directly affect the synthesis of NSDs, which, in turn, affects the glycosylation process and the final product quality. We plan to validate this model-based tool with experimental data from cell cultures conducted using stable and transient transfectants. Even though manufacturing cell lines are stably transfected and clonal, transient transfections are of particular interest for the rapid provision of material for clinical evaluation. It is therefore of paramount importance that the material provided is similar to what would be produced from the final production cell lines. The project will be organised around the following objectives: a) Experimental analysis of the effect of residual glucose and glutamine concentration, ammonia accumulation and culture pH on the availability of NSDs and the variation of glycoforms produced. b) Comparison of glycoforms produced by stable and transiently transfected industrial CHO cell lines. c) Development and validation of kinetic mathematical model for the transient and stable transfectants using the experimental data generated in objective 1 (We have already begun the development of the modelling tool for the effects of glucose availability). d) Computational design and experimental evaluation of process and genetic engineering strategies for narrowing the gap between the glycomic profiles of transient and stable transfectants. [1] Hayter et al., Biotechnol Bioeng 1992, 39:327. [2] Wong et al., Biotechnol Bioeng 2005, 89:164. [3] Trummer et al., Biotechnol Bioeng 2006, 94:1045. [4] Gawlitzek et al., Biotechnol Bioeng 2000, 68:637. [5] Borys, Nature Biotechnol 1993, 11:720.

Monoclonal antibodies (mAbs) are large proteins the body uses to fight many diseases and infections, and hence they are needed in larger and larger quantities. At the moment they are used in minute quantities for mainly diagnostic testing, and hence are very expensive. Most of this cost is due to their dilute nature in the biological fermentation they are produced in, and their separation on very expensive Protein A chromatographic columns which are very selective for the dilute amounts. This proposal wants to develop a very cheap method for purifying these mAbs using a solvent with very small aggregations of surfactants in it (Reverse Micelles-RM) which can selectively separate these mAbs from their original source. We want to study how fast this occurs, how we can use them to purify mAbs, and how we can stop them precipitating on the phase boundary between the two phases (fermentation broth and solvent).

Doctoral Training Partnerships: a range of postgraduate training is funded by the Research Councils. For information on current funding routes, see the common terminology at https://www.ukri.org/apply-for-funding/how-we-fund-studentships/. Training grants may be to one organisation or to a consortia of research organisations. This portal will show the lead organisation only.