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SSPC Impact Case Study: Biologics

Advanced Biopharmaceutical Technologies Spokes Project

Project Lead:

Prof. Gavin Walker, University of Limerick, Prof. Niall Barron, NIBRT, Dr Jonathan Bones, NIBRT, Dr Colin Clarke, NIBRT, Dr Gavin Davey, Trinity College Dublin

Project Partner:

Pfizer, Allergan, Eli Lilly, Janssen, Merck, Biomarin and Genzyme

Areas of Impact:

Economic, Environmental, Societal

Scientific Domains:

Biologics / Drug Product

With the increasing importance of biopharmaceutical manufacturing, initiating the project was a strategic move due to the growth annually in biopharmaceutical sales, representing the fastest growing segment within company pipelines. This project, which concluded in December 2017, included seven of the leading Biopharmaceutical companies in Ireland (Allergan, Biomarin, Eli Lilly, Janssen, MSD, Pfizer, Sanofi-Genzyme) and researchers from two of Ireland’s leading pharmaceutical research Centres SSPC and National Institute for Biopharmaceutical Research and Training (NIBRT). This SPOKE expanded the breath of the SSPC research programme to include large molecules incorporating the expertise and facilities of NIBRT.

The Challenge:

Driven by the need to improve quality and efficiency, this partnership brought together Ireland’s leading scientists and engineers with industry partners to develop innovative single-use disposable systems for bioprocessing. Currently within the biopharmaceutical industry there is widespread utilisation of single-use technologies to ensure that manufacturing facilities can be flexible, cost-effective and constructed with minimal capital investment and timelines. Extractable and leachable (E&L) compounds, arising from the interaction of culture media with disposable components within bioprocessing, represent a major concern for companies and regulators.

The Solution:

This SPOKE and SSPC facilitate requirement through (1) the provision of highly trained scientists and engineers, (2) knowledge exchange and (3) excellent research outputs that directly impact the R&D process within the biopharmaceutical industry.
This overall research underpins the current confidence in the use of disposable technologies for biopharmaceutical manufacture with appropriate awareness and controls. There is currently a wave of investments globally in disposable production facilities and every facility will be heavily scrutinised by the regulatory bodies to ensure that the risks of leachates from plastic components are adequately controlled to ensure that they do not impact production consistency, product quality or interfere with analytical test methods. The research from SSPC spoke team shows that the mechanism of action of bDtBPP impacts cell growth, but not final product quality attributes. However the regulatory scrutiny in this area will intensify as the technology continues to be adopted and applications broaden, particularly as it is now beginning to be adopted in the manufacturing of parenterals, which are much closer to the patient.

The Impact:

This partnership has resulted in the development of advanced technologies to maximize biotherapeutic production, reducing adverse effects of E&L’s and increasing bioengineering capability during cell bioprocess in polymeric disposable systems. SSPC and NIBRT are now leading the way in collaboration across the small to the large molecule sectors augmenting the work of the SSPC research programme, to bridge the gap from the molecule to the material to the medicine.
The project led to improved success of pharma development of novel biologics for therapeutic benefit, and, improved pharma performance, through streamlining of manufacturing, with obvious economic implications passed along to the end user. This has impacted public health and well-being through improved and more cost appropriate pharma products, resulting from streamlined bioprocess and manufacturing methods with associated cost reduction in manufacturing of biologics. Environmentally this could help minimise waste by-product generation and natural resource utilisation during manufacturing.

The industry partners benefit from the project at several levels, for example, identification of possible metabolic pathways affected by E&L from single use bags supplied by different vendors, gaining a deeper insight into the underlying mechanics link between cellular metabolism and CHO cells growth control, biopharmaceutical titer and quality and patient health. Promotion of practical strategies to improve the tolerance of CHO cells against E&L from single use technologies.

The €1.5m in industry/state funding invested in this partnership has already had a direct economic impact with the on-boarding of 7 post-doctoral researchers, enhancing the value and power of Ireland’s R&D program. This collaboration positively impacts the research ecosystem in Ireland, which has a direct impact on existing companies and also strengthens the business case for new Foreign Direct Investment (FDI) to locate in Ireland.

  • Development of a sophisticated method ultra-high-performance chromatography coupled with quadrupole-orbitrap mass spectrometry (UPLC-QToF-MS) to screen the disposable bags from the leading vendors in this field and also assessed first and second generations of the product lines.
  • Deployment of a multi-omic (glycomic, proteomic, metabolomics) approach to analysing cells incubated in media pre-conditioned with bDtBPP to characterise what is involved in the response to this target leachate and to widen understanding of the potential to be impacted by new leachates in the future by the same mechanism.
  • Development of analytical platforms to allow industry to identify and select the most suitable SUBs for cell cultivation according to specific purposes, based on results of extractables and leachable’s (E&Ls) assessments and correlations found with several characteristics of the disposable bags.
  • Development of a detailed cell culture-based assessment of the phenotypic impact of a known leachate on CHO cell behaviour in culture and an understanding of the levels of that compound that may be problematic when using these single-use solutions.
  • Papers published from the project have been well received by the international community. This was evidenced by an invitation to convert a poster presentation into a full oral presentation at the 25th ESACT meeting in Lausanne.
  • First report published on the effects of bDtBPP on biotherapeutic products.
  • Method development for MS analysis of CHO cells cellular N-glycome and
  • Quantitive comparison of glycomes from perturbated and unperturbated cells
    A new metabolic flux control model was developed allowing rapid identification of ongoing metabolic events inside CHO cells and facilitates bioprocess
  • Deeper insight was gained into the underlying mechanics linking cellular metabolism and CHO cells growth control, biopharmaceutical titre and quality and patient health
  • Development of practical strategies to improve the tolerance of CHO cells against E & L from single use technologies.
  • Evaluation of the suitability of Waters PATROL UPLC Process Analysis System for upstream online monitoring of E&L compounds during the bioprocess, which has not been done to date.
  • Demonstration of tangible improvement in new generation material design for SUBs based on reduced negative impact of CHO cell physiology

Publications arising directly from the Spoke

Ta and J. Bones, Development and validation of an ultra-performance liquid chromatography method for the determination of bis(2,4-di-tert-butylphenyl)phosphate and related extractable compounds from single-use plastic films. J. Chromatogr. A. 1492 (2017) 49-54.

Dorival–García and J. Bones. 2017. Monitoring Leachables from Single-Use Bioreactor Bags for Mammalian Cell Culture by Dispersive Liquid-Liquid Microextraction (DLLME) followed by UHPLC-QToF-MS. J. Chromatogr. A. 1512: 51–60.

Dorival–García and J. Bones. 2017. Evaluation of Solvent Systems for Optimized Extractables Studies of Single Use Bioprocessing Solutions. J. Chromatogr. A. 1513: 69–77..N. Dorival-García, S. Carillo, C. Ta, D. Roberts, K. Comstock, S. Lofthouse, E. Ciceri, K. D’Silva, G. Kierans, C. Kaisermayer, R. Hayes, J. Bones. Large-scale assessment of extractables and leachables in single-use bags for biomanufacturing.

Dorival–García, F. Galbiati, R. Kruell, R. Kovasy, K. D’Silva, J. Bones. Identification of additives in different polymer materials from single-use bags used in bioprocessing by accelerated solvent extraction and ultra high-performance liquid chromatography coupled with quadrupole orbitrap high-resolution mass spectrometry.

Carillo and J. Bones. In-depth characterisation of CHO cells N-glycome and their response to bDtBPP exposure.

Kelly PS, McSweeney S, Coleman O, Carillo S, Henry M, Chandran D, Kellett A, Bones J, Clynes M, Meleady P, Barron N. 2016. Process-relevant concentrations of the leachable bDtBPP impact negatively on CHO cell production characteristics. Biotechnol Prog. 32(6):1547-1558.

Hayes JM, Frostell A, Karlsson R, Müller S, Millan-Martin S, Pauers M, Reuss F, Cosgrave E, Anneren C, Davey GP and Rudd PM. (2017) Identification of Fc gamma receptor glycoforms that produce differential binding kinetics for rituximab. Molecular Cell Proteomics doi: 10.1074/mcp.M117.066944

Hayes JM, Wormwald M, Rudd P and Davey GP* (2016) Fc gamma receptors: glycobiology and therapeutics prospects. Journal of Inflammation Research 9, 209-219

McDonald AG, Hayes JM and Davey GP* (2016) Metabolic flux control and glycosylation Current Opinion in Structural Biology 40, 97-103

McDonald AG, Tipton KF and Davey GP* (2016) A knowledge-based system for display and prediction of O-glycosylation network behavior in response to enzyme knockouts. PLOS Computational Biology 12(4):e1004844

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