An advanced bioreactor facility for automated, industry-aligned production of high-value proteins

Lead Research Organisation: University of Kent
Department Name: Sch of Biosciences

Abstract

The production of recombinant proteins is a critically important area in biology, and many proteins are currently produced in bacterial, yeast or mammalian cells and then purified so that their structures and mechanisms can be studied in detail. However, recombinant protein production is also a central part of the biotechnology industry, because many of the most powerful medicines are 'biopharmaceuticals' such as anti-cancer or anti-inflammatory antibodies, cytokines, growth hormones and others. The worldwide biopharmaceutical industry is valued at over $160 billion p.a. and it is growing rapidly. Research into new biopharmaceutical drugs and expression systems is a major priority in the UK.

Many academic groups work closely with Biotechnology companies to develop new cell lines with enhanced properties, but whereas companies use sophisticated equipment to grow cells and produce proteins in large amounts, such systems are rarely available to academic groups. This means that there is a significant mismatch in efforts, and cell growth protocols developed by academics cannot be simply adopted by their industrial partners. Instead, new growth and protein expression protocols have to be developed from scratch.

In this application we propose to purchase an ambr250 bioreactor system, which allows highly automated growth of bacterial or mammalian cells at very high density. The bioreactor volumes are only 100-250 ml but it has been shown that the results are highly scalable; in other words, growth protocols developed using this system almost invariably work when used used in much larger reactor vessels. The system will be installed in a purpose-designed unit and used in two broad types of project:

(i). Groups working on biotechnology projects, where the aim is to produce high-value proteins such as biopharmaceuticals, will benefit from the equipment because they will be able to carry out research under industry-mimicking conditions. This will enhance progress in a number of important projects at Kent and elsewhere, and the equipment will also be used to enhance work in a new 'Global Challenges' research project involving Kent, Imperial College and University College London. This project aims to enable countries in South East Asia to produce their own biopharmaceuticals, most of which are simply too expensive for the vast majority (over 90%) of patients.
(ii). Groups working on more fundamental areas of science, where large quantities of recombinant protein are required, will benefit because the ambr250 system can produce cells at high densities and target proteins are often produced at concentration of over 1 gram per litre of culture - easily enough for most research projects.

The proposed facility will be used by the applicants and also made available to external users. It will be unique among academic Institutions in the UK: UCL is the only University in possession of an ambr250 system and this is not available to external users. The primary reason for the low presence of this equipment in academia is the cost of the bioreactors, which are expensive and single-use. To circumvent this problem and ensure availability to the academic community, the University of Kent has committed a total of £80,000 for the sole purpose of providing bioreactors free of charge over the first four years of the facility's existence. This will provide a huge boost to uptake by external users and we predict that many groups will be able to capitalise on the system's exceptional performance.

Technical Summary

This application aims to establish a new facility for high-density growth of bacterial and mammalian cells that produce recombinant proteins in large amounts. The main driver is to develop facilities that mimic those used by industry for biopharmaceutical production; most academic groups use relatively simple culture facilities whereas industrial groups use more automated systems that generate far higher cell densities. There is a clear mismatch between most academic groups and their industrial collaborators. We propose to purchase an ambr250 bioreactor system (Sartorius) that is capable of highly automated fed-batch growth of bacterial OR mammalian cells. It is the standard development tool in biotechnology companies, and the availability of the system will enable the applicants to align their research much more closely with the efforts of a range of industrial collaborators. The system will also be made available to external users.

A second important application is the production of large quantities of recombinant proteins for structural and other analyses; while the bioreactors are only 100-250 ml in volume, the system is capable of producing proteins at levels of over 1 gram per litre of culture.

The system will provide a major boost to research in IB Catalyst, sLOLA, GCRF and other research projects in the applicants' groups. However, we will make the facility available to external users and we firmly predict that a wide range of groups, particularly those working in the bioprocessing community, will benefit. A major factor preventing wider academic use of amor systems is the high cost of single-use bioreactors, but to circumvent this problem, Kent has committed £80,000 to purchase bioreactors over 4 years. Most will be provided free of charge to users outside Kent.

The facility will be unique in the UK academic community; only one other ambr250 system is present in UK Universities and it is not available to external users.

Planned Impact

The aim of this programme is to develop a state of the art facility for recombinant protein production, based in the School of Biosciences at the University of Kent. The facility will be centred around an ambr250 advanced bioreactor system which can be used for high-density growth of bacterial or mammalian cells, and the facility will be used for for the production of (i) biopharmaceuticals and (ii) vaccine antigens, and (iii) diverse recombinant proteins for structural and other analysis.

This project will be of direct benefit to a wide cross section of academic groups, and will have a significant positive effect on a range of academic-industry collaborations..

A. Impact on academic research groups.

The impact on academic research should be high. Numerous groups work in the field of 'bioprocessing' and a large academic community has been generated through programmes such as BRIC and the IB Catalyst. The applicants are familiar with nearly all of the groups and it will be straightforward to involve a large number of them in this project, particularly when the bioreactors are provided free of charge for initial work. Equally, many groups struggle to produce recombinant proteins in sufficient quantities for structural analysis and the ambr250 system is capable of producing very high levels of protein in many studies. The facility will be of significant interest to such groups and the availability of bioreactors at no cost over 4 years (as part of Kent's Institutional commitment) will enable a range of groups to use this facility.

B. Impact on industry and society.

This is expected to be unusually high. Bioprocessing research is a major priority among several funding agencies, especially BBSRC, and numerous groups collaborate with industry to develop new bacterial and mammalian cell expression platforms with increased capabilities. However, most academic groups lack the equipment to carry out research under conditions routinely used by industry, and this is clearly a barrier to progress in many cases. The availability of this system will enable a number of groups to characterise and optimise their expression studies in a manner that will be clearly relevant to the needs of their industrial collaborators - nearly all of whom use ambr systems. The UK houses some of the world's most prominent biopharmaceutical companies, many of whom already collaborate with one or more of the applicants. The accelerated development of new expression, cultivation, and downstream processing systems is certain to be of interest to such companies, as are product analysis data on the proteins that the new facility generates.

C. Impact overseas.

A new RCUK £4.9 million Global Challenges Research fund project, led by Robinson, brings together highly-rated UK groups in order to develop effective new platforms and processes for the production of biopharmaceuticals. The programme aims to enable Thailand and other SE Asian countries to produce their own target molecules, because the vast majority of patients in those countries have no access to key anti-cancer and other biologics due to their high costs. The availability of the proposed Facility will clearly enhance progress in this project because the research will be more geared to the pilot-scale growth systems used overseas. Moreover, it is undoubtable that the establishment of integrated production processes during this project will be of interest to UK companies and academics, who are very likely to adapt elements of the programme for their own uses.
 
Description This is an equipment grant that provided an ambr250 automated bioreactor. The advanced bioreactor is used for recombinant protein production and it has been set up and tested. The manufacturer, Sartorius, has helped to train staff in its use and we are now ready to interact with users from other Institutions to develop the facility.
Exploitation Route A range of academic and industrial collaborators have expressed an interest in using this facility
Sectors Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description The facility has been used by University of Kent groups and is fully operational. However, attempts to develop its use by academic and industrial groups were prevented by the Covid-19 pandemic. The equipment is housed within the School of Biosciences at the University of Kent and we are finally able to host visiting groups, so the unit can now be made available to external users.
First Year Of Impact 2020
Sector Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
 
Description Bioscience Animal Health group (BIS) 
Organisation Bioscience Animal Health Group
Country Thailand 
Sector Private 
PI Contribution BIS are one of Thailand's largest distributors of veterinary products including animal vaccines. They are working with our consortium to assist in our porcine vaccine production project.
Collaborator Contribution BIS currently distribute imported vaccines against pig viruses, but are keen to help us produce new vaccines and have offered to market the vaccines when they have been validated. They are planning to market our vaccine once it is approved by the Thai FDA and have offered to fund safety trials in Thailand.
Impact BIS are now heavily involved in the planning of the project and we have agreed that they will have full rights to market the vaccine in Thailand if the efficacy trials are successful
Start Year 2017
 
Description Mahidol University PhD secondment scheme 
Organisation Mahidol University
Country Thailand 
Sector Academic/University 
PI Contribution We are keen to maintain the collaborations built up with Thai centres during this award, and have agreed that several PhD students in our BBSRC 'SoCoBio' Doctoral Training Partnership will undertake their 3-month placements (PIPS) with collaborators in Mahidol University.
Collaborator Contribution Mahidol will host several phD students per year and waive all fees.
Impact Secondment scheme has just started this year
Start Year 2022
 
Description Sartorius 
Organisation Sartorius
Department Sartorius Stedim Biotech
Country France 
Sector Private 
PI Contribution We are setting up a protein expression facility using the equipment purchased on a BBSRC ALERT grant. Thai and UK appointees on the GCRF grant are receiving training in the use of the equipment.
Collaborator Contribution Sartorius supplied the equipment and are helping to establish a facility; they have also provided consumables as part of a collaborative project on optimising the setup
Impact Collaboration is proceeding smoothly and we have submitted a manuscript for publication that includes authors from Sartorius.
Start Year 2018
 
Description Syngenta 
Organisation Syngenta International AG
Department Syngenta Ltd (Bracknell)
Country United Kingdom 
Sector Private 
PI Contribution Syngenta are using E. coli strains developed during this award to develop new methods of expressing high-value recombinant proteins.
Collaborator Contribution The University of Kent + University of Birmingham researchers developed 'TatExpress' strain that export folded proteins to the periplasm in high amounts. These strains and the relevant vectors have been transferred to Syngenta for tests.
Impact Research is ongoing
Start Year 2019
 
Description Vanvimon Saksmerprome 
Organisation National Center for Genetic Engineering and Biotechnology (BIOTEC)
Country Thailand 
Sector Public 
PI Contribution Dr Saksmerprome (BIOTEC) is working on vaccines against shrimp viruses. She is working with University of Kent PIs to develop new systems to combat infections and the Robinson group is developing algal strains that express anti-viral RNAs. Algal strains expressing these molecules have been sent to Thailand and are undergoing efficacy trials in shrimp larvae.
Collaborator Contribution The Kent group is developing new methods to produce anti-viral molecules that can be used to prevent viral infections that cause massive losses in the Thai shrimp industry. She is now an 'associate partner' in the GCRF consortium. Dr Saksmerprome has recently been awarded a Newton Advanced Fellowship to work with Prof Robinson's group, significantly strengthening the collaboration. Very recently, Prof Robinson, Dr Saksmerprome and another Thai PI have been awarded a 3-year Royal Society International Collaboration Award to develop an expanded research programme.
Impact The Newton Advanced fellowship (£74,000) and Royal Society ICA (£225k) described above
Start Year 2017
 
Description ESACT annual conference, 2020, Ashford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Plenary talk at the annual ESACT conference on cell culture systems
Year(s) Of Engagement Activity 2020
 
Description Presentation at International Veterinary Vaccine Network conference, Hanoi, December 2020 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Policymakers/politicians
Results and Impact Dr peera Jaru-Ampornpan (PI on this grant) gave an invited presentation at the IVVN network conference 'Vaccines for Swine'
Year(s) Of Engagement Activity 2020