Six new BIVs awarded

BioProNET has recently awarded six business interaction vouchers:

Industry biotechnology for the production of hyaluronic acid from Streptococcus equi: understanding hyaluronic acid expression
Garry Blakely, University of Edinburgh working with Hyaltech
We wish to understand how the production of hyaluronic acid in Streptococcus equi changes during large-scale fermentation. To acheive this, we will study the genetic mechanisms that regulate expression in order to to yield quantitative improvements in production.

Exploring the feasibility of nonlinear acoustic detection technique for online bioprocess monitoring
Sourav Ghosh, Loughborough University and the Centre for process innovation
This project proposes to evaluate whether a nonlinear acoustic detection technique could offer a sensitive yet fast and reliable alternative to current industrial standard analytical methods (such as immunoassay, HPLC/UPLC).

Supercritical fluid processing to improve the stability and delivery of low dose biopharmaceuticals
Helen Philippou, University of Leeds in collaboration with Crystec
Crystec has innovative supercritical fluid technology that can be used can process larger molecules to form room-temperature stable particles. This project will be used to evaluate whether supercritical fluid technology can be used on a micro-scale to successfully process low-dose biomolecules..

A collaboration to engineer a novel protein nanopore for single molecule DNA sequencing applications
Michael Plevin, University of York working with Oxford Nanopore Technologies
Oxford Nanopore Technologies produce phone-sized DNA sequencer, in which DNA is sequenced as it passes through a motor protein-controlled nonopaore. We will determine whether recmoninant proteins native to heat-loving microbes can be engineered to improve the DNA reading capabilities of the sequencer.

Developing a novel fluorescence-based biopharmaceutical quality control technology
Christopher Pudney, University of Bath in collaboration with Bath ASU
We will develop a novel technology to accurately perform quality control on biopharmaceuticals. The technology is based on the quantification of the fluorescence edge shift phenomenon giving a library of spectroscopic fingerprints for different biopharmaceuticals, which accurately quantifies subtle changes to protein structure.

Predictive tools for folding-supportive sequence design spaces
Tobias von der Haar, University of Kent working with UCB Celltech
This project aims to test if it is possible to combine high yield and high activity during the production of cell-derived, protein-based pharmaceuticals. We will test whether tools to boost yield can be adapted to improve production of novel-format antibodies.

See below for BIVs that we have awarded in previous calls:

Collaboration on a bio-process to optimize the yields and to characterize and test unique exopolysaccharides from two microalgal strains
Kevin Flynn, Swansea University; John Dodd, Algaecytes
Exopolysaccharides are high-molecular-weight polymers that are composed of sugars and are secreted by microalgae into the surrounding environment to protect them against stress. Exopolysaccharides  have biological activity against viruses, antioxidant activity and also can help to improve the immune system in mammals. The aim of the project is to grow two microalgae strains in large volumes and concentrate the exopolysaccharides by the use of a novel reverse osmosis technique (a new bio-process). Samples will then be chemically characterized using FTIR/EDAX techniques. The exopolysaccharides will be screened empirically for bioactivity to seek commercial application of a biologic product.

Enhancing cell growth to allow selection of biopharmaceutical-producer cell lines with favourable properties
Lisa Swanton, Faculty of Life Sciences,University of Manchester; Alasdair Robertson, SAL Scientific
Protein-based therapeutics represent a strategically important and rapidly expanding component of the industrial biotechnology sector, and are typically manufactured via bioprocessing using mammalian host cells engineered to produce high levels of the therapeutic protein. A major limitation is the certainty of ensured survival and selection of the most highly productive cell lines. We will test whether the addition of novel supplements that enhance cell growth/survival during cell line selection of stable cell lines (CHO S cells, transfected to express “difficult-to-express” variants of erythropoietin) enhances the recovery of productive clones in greater numbers and with greater specific productivity of recombinant protein.

Monitoring of host cell proteome expression during bioprocessing of CHO cells expressing recombinant proteins
Martin Michaelis, School of Biosciences, University of Kent; Michael Hutchins, TotalLab
When recombinant Chinese hamster ovary (CHO) cells expressing biotherapeutic monoclonal antibodies are cultured and the cell culture fluid harvested, both antibody product and CHO host cell proteins (HCPs) are present necessitating a multi-step purification procedure. It is a regulatory requirement to determine the amounts of these HCPs in the therapeutic product and current methods to determine this include using 2-dimensional gel electrophoresis (2D-PAGE). Here we will use 2D-PAGE to analyse the HCP proteome during culture of CHO cells, and potentially during early purification steps, and then apply new software developed by TotalLab to determine those HCPs present and their relative abundance. These data will provide the basis for risk-based assessment of the presence and relative concentration of specific HCPs to provide better control of HCP amounts and improved assurance of recombinant protein product quality. We will also assess if the new software is more sensitive, accurate and rapid in defining the HCP profile than current technologies.

Design consultation and testing of a membrane photobiorector suitable for advanced biologic production from micro algae
Mike Allen, Plymouth Marine Laboratory, Joe McDonald, Varicon Aqua Solutions
The production of high value biologics in plankton is uneconomical, primarily due to the demands of mixing and harvesting low biomass concentrations from large volumes. This  issue is almost entirely negated by the use of bioreactors that use a biofilm-membrane design, in which the biomass is retained within a high-density, low-volume system. To our knowledge, however, there is no commercially available membrane system suitable for the growth of algae as a biofilm. This project will develop a prototype membrane photobioreactor for high value biologic production from microalgae — bench top size; about 5l volume — and will assess biologic production capability using a reference algal system: a genetically modified strain of Phaeodactylum tricornutum expressing the Venus Yellow Fluorescent Protein.To achieve this scientists from Varicon Aqua Solutions (VAS) will work alongside researchers from PML, advising on photobioreactor design and production assessment, fabricating the photobioreactor, incorporating existing VAS control and lighting systems, providing nutrients, and analyzing data. Funds from BioProNET will cover consumables and parts as well as investigator costs

Evaluating the use of Raman Spectroscopy to determine topological isoforms of plasmid DNA
Lorna Ashton, Department of Chemistry, Lancaster University, who will be partnering with Cobra biologics.
The use of plasmid DNA as a direct gene therapy product, and as a critical starting material for transient viral vector production, is a rapidly growing global market. Current analytical methods for determining plasmid tertiary structure are highly invasive, requiring a high degree of sample preparation. Raman spectroscopy is non-invasive, providing near real-time information on therapeutically relevant biological molecules (proteins, nucleic acids and viruses)
We will test if Raman spectroscopy can identify different plasmid DNA topologies, and if the methodology is able to provide quantitative data, compared with standard orthogonal analytical approaches, on the various plasmid isoform levels. A better knowledge of DNA topology will aid the formulation and in-process development of biological molecules.

Production of therapeutic and industrial proteins in microalgae
Anil Day, Faculty of Life Sciences, University of Manchester, who will be collaborating with Protein Technologies.
Proteins are natural catalysts and include antibodies that can bind a diverse array of molecules including those present in viruses, bacteria and cancer cells. They have multiple applications in industrial biotechnology by speeding up chemical reactions, detecting and visualizing molecules in cells, and treating diseases. Modern manufacture of therapeutic and industrial proteins for specific applications requires a well characterized, reliable and safe host cell to manufacture proteins. In this project we will use microalgae, which provide an attractive new system, which is low-cost, safe and sustainable, to make proteins with applications in healthcare.

Exploiting advanced electron microscopy to optimise protein and biologic expression platforms
Corinne Smith, University of Warwick and Colin Robison, University of Kent will be working with Jeol UK
Modern bioscience and industrial biotechnology processes depend on expression and export of recombinant proteins — such as biologics — in a very small number of systems. E. coli is a popular expression system because it is easy to work with and cheap to run; Surprisingly, we know very little about the distribution and shape of the structural elements in the E. coli organism which drive protein expression and export to the periplasm. A better understanding of these processes will facilitate better bioprocessing technologies.
We propose to work with Jeol U.K. to exploit advanced electron microscopy technology to image organisms such as E. coli in fine detail using zero-loss cryo-electron tomography and direct electron detection. This will provide ultra-high resolution data on the effects of high-level recombinant protein production in terms of:
(i) Distribution of any insoluble protein aggregates (inclusion bodies, which will hamper expression and export).
(ii) Effects on levels and distribution of protein export systems, using immunogold electron microscopy – this is important because it will help us to understand if over-expressed protein export system is properly localised.
(iii) Generalised effects on membrane structures.

Development of a crossflow filtration dynamic flux control system to reduce cell harvest time
Yuhong Zhou, Department of Biochemical Engineering, University College London is working on a project with BioProControl Tech.
Crossflow filtration is one of the key operations for cell harvest, particularly when cells are
shear sensitive such as infectious cells and cell therapy products. A short operation time is desirable
but often achieved by using conservative fluxes (to prevent fouling) with a large membrane area hencehigh cost at large scale processes. A dynamic control system that balances the flux
and the fouling over the entire operation has the potential to shorten the operating time. A dynamic control system combining model-based control and feedback control strategies is proposed. Its feasibilitywill be tested in E. coli cell harvest.

Rapid processing to recover high value microbial by-products
Paul Clegg and Joe Tavacoli, School of Physics & Astronomy, University of Edinburgh are collaborating with Recyclatech.
Recyclatech employs mycolic-acid producing bacteria in industrial biotechnological processes that generate large volumes of spent medium containing effective biosurfactants and bacteria that can be utilized in bioremediation of oil contaminated soils and aqueous matrices. The biosurfactants can be used for home-care, pharmaceutical and other products.
The project aims to develop a rapid recovery system, using extensive expertise available at the University of Edinburgh, for these commodities by forming droplets that hold the bacteria and cell-associated biosurfactants. The bio-material can then be facilely removed by a skimming process. Better and/or more cost-effective ways to recover spent medium will benefit many bioprocessing applications.

A pilot study to improve the expression of a Clostridium difficile toxin-based fragment in E.coli
Tarit Mukhopadhyay and Michael Sulu, Deptment of Biochemical Engineering, University College London will be partnering with Public Health England.
Clostridium difficile
is a major cause of hospital and community-acquired infections. This bacterium produces toxins in the gut that lead to diarrhoea and gut inflammation. This is a serious disease and can be fatal. Public Health England is developing an antibody-based therapy that will neutralize these toxins and halt infection symptoms. To do this, toxin-derived antigens are required to produce ovine antibodies and in this project we seek to express these antigens in harmless strains of E. coli. This project will investigate antigen expression and manufacturing protocols for this new treatment.

Evaluating enhancement of Secretion for Recombinant Proteins in CHO cells via overexpression of 7SL RNA
Bob White, University of York and Cobra Biologics
Secretion of therapeutic monoclonal antibodies for clinical production is a highly desirable requirement for a scalable manufacturing process. In some production systems, bottlenecks in secretion can limit antibody production. Signal recognition particle (SRP) is an essential component involved in protein secretion. It consists of six proteins assembled around 7SL RNA, the essential scaffold of SRP. We will test if the overexpression of 7SL RNA can increase the amount of a therapeutically-relevant monoclonal antibody secreted by a stably-transfected cell line. We will also examine the intracellular trafficking of the antibody to better understand the secretion bottleneck.

Initial development of novel product concepts with unique pharmacokinetic characteristics Randall Mrsny, University of Bath and Arecor
Arecor has developed technologies for stabilising therapeutic proteins. Some of these have the potential to modify the pharmacokinetic profile of therapeutic protein or peptide (biopharmaceuticals) following subcutaneous (SC) injection. University of Bath has developed an in vitro technique for modelling interactions between a biopharmaceutical and hypodermis components that mimics SC injection events. This information describes bioavailability outcomes in humans and we believe can also be deconvoluted to acquire pharmacokinetic parameters. This project will combine respective strengths of Arecor and Bath to generate proof-of concept data on selected therapeutic proteins, demonstrating the ability to produce a desirable pharmacokinetic profile. A better understanding of the key parameters that control the pharmacokinetic properties of injectable biopharmaceuticals will improve (for example, by allowing rationale design based on the formulation) the process of producing stable, functionally active biopharmaceuticals that are commercially useful.

If you would like to submit an application for a business interaction vouchers, please see the ‘funding’ tab.