Development of a microfluidic bioanalytical device for the characterization of cellular therapeutic products
Lead Research Organisation:
University College London
Department Name: Biochemical Engineering
Abstract
The development of cell and gene therapies, as well as their manufacture at production scale relies on the capacity to expand mammalian cell cultures traceably, reproducibly and cost effectively. To achieve this aim, one needs to go beyond the automation of bioprocesses and the development of novel single-use technologies. In particular, for effective and high quality bioprocessing, robust analytical methods for process monitoring, as well as for the characterization of the final product are of paramount importance. Online measurement in bioreactors is often limited to a few key process variables, including dissolved oxygen (DO), temperature, and pH. However, to attain precise process control, it is necessary to monitor multiple metabolites in the complex cell culture media. This information can then be employed to optimize feeding rates, and ultimately obtain higher cell yields of better quality cells. For mammalian cell culture, there are many challenges involved in applying existing process analytical techniques (PATs) for the monitoring of key metabolites such as glucose, lactate or glutamine. Current strategies for culture media characterization are limited: they use sensors which are analyte-specific, requiring one probe for each analyte, and therefore significantly increase the cost of the process. Furthermore, probes are invasive, and enzymatic sensors, widely used for metabolic screening, degrade during long processes, affecting the accuracy, and limiting the sampling regime.
New approaches to process analytical technologies will be required to meet the challenges imposed on the industry with the advent of personalized medicine: single-use technologies, inline (or at-line) monitoring of key culture variables, the integration of data acquisition with high throughput adaptive algorithms to increase reproducibility and reduce cost through effective process automation. ; Spectrophotometric techniques allow non-invasive monitoring of culture media composition as well as the characterization of intracellular metabolism and general cell quality. New microfabrication technologies have enabled microfluidics and micro-electro-mechanical devices (MEMs) to be developed for the handling of microscale liquid volumes, offering new approaches to sample withdrawal, separation and preparation prior to chemical processing or screening. Much work is being carried in the miniaturization of traditional processes, such as liquid chromatography for the processing of microfluidic samples due to the reduction of reagents and improvement of flow control. Furthermore, immobilization techniques allow for electrochemical or chemoluminescent screening of a variety of relevant analytes. Microfluidic sample handling can also be used for cell withdrawal, concentration and exposure, for the characterization of cellular features such as gene expression or internal metabolism. The 'lab-on-chip' capabilities of microfluidic and microelectromechanical can be used along with traditional spectroscopic techniques used in metablomics such as, MS, UV-Vis, MIR, fluorescence and other emerging techniques such as Raman or combined Raman/NIR spectroscopy. Of these techniques, Raman spectroscopy has the capacity to detect virtually any constituent of cell media with little or no sample derivatization, given its high signal-to-noise ratio with aqueous samples.
The aim of this research is to design, develop and integrate a bioanalytical tool for the characterization of automated T-Cell and mesenchymal stem cell expansion processes. The samples will be interrogated spectroscopically and electrochemically to obtain key process information on the chemical environment and cellular quality of the culture.
New approaches to process analytical technologies will be required to meet the challenges imposed on the industry with the advent of personalized medicine: single-use technologies, inline (or at-line) monitoring of key culture variables, the integration of data acquisition with high throughput adaptive algorithms to increase reproducibility and reduce cost through effective process automation. ; Spectrophotometric techniques allow non-invasive monitoring of culture media composition as well as the characterization of intracellular metabolism and general cell quality. New microfabrication technologies have enabled microfluidics and micro-electro-mechanical devices (MEMs) to be developed for the handling of microscale liquid volumes, offering new approaches to sample withdrawal, separation and preparation prior to chemical processing or screening. Much work is being carried in the miniaturization of traditional processes, such as liquid chromatography for the processing of microfluidic samples due to the reduction of reagents and improvement of flow control. Furthermore, immobilization techniques allow for electrochemical or chemoluminescent screening of a variety of relevant analytes. Microfluidic sample handling can also be used for cell withdrawal, concentration and exposure, for the characterization of cellular features such as gene expression or internal metabolism. The 'lab-on-chip' capabilities of microfluidic and microelectromechanical can be used along with traditional spectroscopic techniques used in metablomics such as, MS, UV-Vis, MIR, fluorescence and other emerging techniques such as Raman or combined Raman/NIR spectroscopy. Of these techniques, Raman spectroscopy has the capacity to detect virtually any constituent of cell media with little or no sample derivatization, given its high signal-to-noise ratio with aqueous samples.
The aim of this research is to design, develop and integrate a bioanalytical tool for the characterization of automated T-Cell and mesenchymal stem cell expansion processes. The samples will be interrogated spectroscopically and electrochemically to obtain key process information on the chemical environment and cellular quality of the culture.
Planned Impact
The IDC has a proven track record of delivering impact from its research and training activities and this will continue in the new Centre. The main types of impact relate to: (i) provision of highly skilled EngD graduates; (ii) generation of intellectual property (IP) in support of collaborating companies or for new venture creation; (iii) knowledge exchange to the wider bioprocess-using industries; (iv) benefits to patients in terms of new and more cost effective medicines, and (v) benefits to wider society via involvement in public engagement activities and encouraging future generations of researchers.
With regard to training, the provision of future bioindustry leaders is the primary mission of the IDC and some 97% of previous EngD graduates have progressed to relevant bioindustry careers. These highly skilled individuals help catalyse the development and expansion of private sector innovation and biomanufacturing activity. This is of enormous importance to capitalise on emerging markets and to create new jobs and a skilled labour force to underpin the UK economy.
In terms of IP generation each industry-collaborative EngD project will have direct impact on the industry sponsor in terms of new technology generation and improvements to existing processes or procedures. Where substantial IP is generated this has the potential to lead to spin-out company creation and job creation with wider UK economic benefit. IDC research has already led to creation of two UCL spin-out companies focussed on the emerging field of Synthetic Biology (Synthace) and novel nanofibre adsorbents for improved bioseparations (Puridify). Once arising IP is protected the IDC also provides a route for wider dissemination of project outputs and knowledge exchange available to all UK bioprocess-using companies. This occurs via UCL MBI Training Programme modules which have been attended by more than 1000 individuals from over 250 companies to date.
The majority of IDC projects address production of new medicines or process improvements for pharmaceutical or biopharmaceutical manufacture which directly benefit healthcare providers and patients. Examples arising from previous EngD projects have included: engineered enzymes used in the synthesis of a novel pharmaceutical; early stage bioprocess development for a new meningitis vaccine; redevelopment of the bioprocess for manufacture of the UK anthrax vaccine; and establishment of a cGMP process for manufacture of a tissue-engineered trachea (this was subsequently transplanted into a child with airway disease and the EngD researcher was featured preparing the trachea in the BBC's Great Ormond Street series). Each of these examples demonstrates IDC impact on the development of cost-effective new medicines and therapies. These will benefit society and provide new tools for the NHS to meet the changing requirements for 21st Century healthcare provision.
Finally, in terms of wider public engagement and society, the IDC has achieved substantial impact via involvement of staff and researchers in activities with schools (STEMnet, HeadStart courses), presentations at science fairs (Big Bang, Cheltenham), delivery of high profile public lectures (Wellcome Trust, Royal Institution) as well as TV and radio presentations. The next generation of IDC researchers will be increasingly involved in such outreach activities to explain how the potential economic and environmental benefits of Synthetic Biology can be delivered safely and responsibly.
With regard to training, the provision of future bioindustry leaders is the primary mission of the IDC and some 97% of previous EngD graduates have progressed to relevant bioindustry careers. These highly skilled individuals help catalyse the development and expansion of private sector innovation and biomanufacturing activity. This is of enormous importance to capitalise on emerging markets and to create new jobs and a skilled labour force to underpin the UK economy.
In terms of IP generation each industry-collaborative EngD project will have direct impact on the industry sponsor in terms of new technology generation and improvements to existing processes or procedures. Where substantial IP is generated this has the potential to lead to spin-out company creation and job creation with wider UK economic benefit. IDC research has already led to creation of two UCL spin-out companies focussed on the emerging field of Synthetic Biology (Synthace) and novel nanofibre adsorbents for improved bioseparations (Puridify). Once arising IP is protected the IDC also provides a route for wider dissemination of project outputs and knowledge exchange available to all UK bioprocess-using companies. This occurs via UCL MBI Training Programme modules which have been attended by more than 1000 individuals from over 250 companies to date.
The majority of IDC projects address production of new medicines or process improvements for pharmaceutical or biopharmaceutical manufacture which directly benefit healthcare providers and patients. Examples arising from previous EngD projects have included: engineered enzymes used in the synthesis of a novel pharmaceutical; early stage bioprocess development for a new meningitis vaccine; redevelopment of the bioprocess for manufacture of the UK anthrax vaccine; and establishment of a cGMP process for manufacture of a tissue-engineered trachea (this was subsequently transplanted into a child with airway disease and the EngD researcher was featured preparing the trachea in the BBC's Great Ormond Street series). Each of these examples demonstrates IDC impact on the development of cost-effective new medicines and therapies. These will benefit society and provide new tools for the NHS to meet the changing requirements for 21st Century healthcare provision.
Finally, in terms of wider public engagement and society, the IDC has achieved substantial impact via involvement of staff and researchers in activities with schools (STEMnet, HeadStart courses), presentations at science fairs (Big Bang, Cheltenham), delivery of high profile public lectures (Wellcome Trust, Royal Institution) as well as TV and radio presentations. The next generation of IDC researchers will be increasingly involved in such outreach activities to explain how the potential economic and environmental benefits of Synthetic Biology can be delivered safely and responsibly.
