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Next-generation biopharmaceutical downstream process

Periodic Reporting for period 1 - nextBioPharmDSP (Next-generation biopharmaceutical downstream process)

Reporting period: 2015-03-01 to 2016-08-31

The scope of the project is the optimization of downstream processing (DSP) for the production of Biopharmaceuticals. Biopharmaceuticals have been successfully used as efficient therapeutic drugs for many pathophysiological conditions since the first recombinant product, insulin, was approved in 1982. Despite its efficacy, accessibility is still limited due to extremely high costs. In the production chain, capturing and purifying still represents a major bottleneck. Consequently, improvements in this area produce substantial cost reductions and expand patients’ accessibility to highly efficient drugs. Another aim of this action is to cope with the changing manufacturing demands, by lowering its environmental footprint and moving to more sustainable technologies.
The project’s main objective is to implement a fully integrated manufacturing platform based on continuous chromatography, in combination with single-use disposable techniques for all unit operations of DSP sequence for biosimilar monoclonal antibodies and derivatives thereof on pilot or small production scale together with incorporation of advanced analytical tools. The action encompasses the entire DSP sequence.
For the activities connected to primary separation of product from cells alternative technologies are being implemented, such as flocculation or alternating / tangential flow filtration (ATF / TFF). The expected outcome is a reduction in the size and number of downstream unit operations and the elimination of centrifugation. Data we generated during the development of flocculation already showed that the process shows significant cost benefits.
Alternative approaches to the batch process for the capture step, such as continuous chromatography, are being evaluated in order to improve the efficiency and lower the need for expensive resin volume. It was already shown, that resin can be utilized to a much higher extent and productivity of this step can be improved significantly. Those inputs were used for designing a novel disposable continuous chromatography system, which is being developed as a part of this project. In addition non-chromatographic approaches are being evaluated as a replacement of first chromatography step. Here continuous precipitation is developed together with prototypes of disposable flow-through reactors.
Since single-use disposable technologies can substitute the extensive use of resources (water) and significantly reduce the overall utility needs, the whole DSP sequence is being developed on disposable technology, which includes evaluation of pre-packed disposable columns, various membrane adsorbers and flow-through templates, where different devices are being interconnected.
Continuous manufacturing requires novel approaches for process control, therefore advanced analytical tools are being developed and implemented, enabling the monitoring of different quality attributes in real time.
The underlying strategy for process development, which aligns the different molecular formats to the different unit operations and developed process schemes, will be established with the latest technology in terms of high throughput process development (HTPD) approaches together with enhanced model based process development. This enables modern quality by design (QbD) concepts and novel process analytical technology (PAT).
Main work performed within 18M of the project is described below.

WP1 - Management:
- All the agreements were signed in time (Grant Agreement and Consortium Agreement).
- Consortium boards were established and are functioning normally (General Assembly, Scientific/Executive Board, Innovation-Exploitation Board).
- Meeting structure was established (kick-off meeting and 3 regular consortium meetings organized; regular management overview meetings; regular scientific WP meetings; specific meetings).
- Tools for efficient administration were implemented (project sharepoint for sharing documents and meeting minutes; website; templates; regular TCs).
- Financial coordination is implemented (4PM software for tracking the financial data for all the partners; overview meetings).
- IP coordination (intellectual property policy was prepared and confirmed; IP capture: IP manager and Innovation/Exploitation board actions).

WP2 - Primary Separation
- Starting material was being prepared as needed.
- Continuous perfusion bioprocesses based on TFF, ATF and centrifugation was developed and tested (ATF/TFF showed best performance).
- Primary separation for fed batch production was developed and optimized by the use of different flocculants in combination with depth filtration (pDADMAC best performing).

WP3 - Capture Step
- Starting material was prepared.
- For alternative batch process on protein A, different loading strategies based on computational modelling were developed and verified experimentally.
- For continuous chromatography process development protein A resin and buffer screening studies were performed together with cleaning efficiency studies. Inputs were used and multi-column continuous chromatography process was developed on more resins.
- Ion-exchange periodic counter-current chromatography (IEX-PCCC) process was developed based on mechanistic modelling and optimization is ongoing.
- For integrated precipitation and IEX-PCCC process a HT precipitation platform was established and prototype reactors for continuous precipitation were developed and tested.

WP4 - Continuous chromatography equipment
- Exploration study was prepared and design around due to patent situation was performed.
- New Product Development Specification (NPDS) document and Process and Instrumentation Diagram (P&ID) were prepared and confirmed.
- 3D mechanical concept has been drafted and user interface was designed and is now being developed.

WP5 - Disposable DSP
- Starting material was prepared.
- Continuous low pH viral inactivation step is being developed.
- mAb flow-through template was evaluated and performance was confirmed. Further optimization will follow.
- Performance of pre-packed columns is being evaluated and anion exchange FT step was developed on membrane adsorbers.
- Disposable UF/DF device was evaluated and SPTFF approaches are being evaluated for continuous concentration of product.

WP6 - Advanced analytical tools
- Detailed definition and description of requirements for AAT was performed.
- Classical analytical methods, which are used as regular analytics and to evaluate the performance of novel AAT were established.
- Protein concentration determination - UV/Vis & FTIR PLS-based peak deconvolution calibrated models were developed and combined with the Flow-VPE device for product breakthrough monitoring.
- Quartz crystal microbalance is being evaluated as a tool for mAb content determination.
- Focused beam reflectance measurements have been successfully used to monitor protein particle formation during UF/DF and flocculation.
- Lab-on-a-chip prototype was developed and manufactured and can be used as an at line tool for monitoring protein particles in the low µm range.

WP7 - Final process
- Start of WP planned for M37, some preliminary discussions started on defining the final setup.

WP8 - Dissemination and exploitation
- Project logo and webpage were created.
- Stakeholders board was finalized and main players were approached
The final expected result of the project is implementation of a fully integrated manufacturing platform based on continuous chromatography, in combination with single-use disposable techniques for all unit operations of DSP on pilot or small production scale together with incorporation of advanced analytical tools.

The project will contribute to the Key Enabling Technologies (KETs):
-The action is expected to develop technology building blocks representing real progress towards better health (societal challenge 1). In fact the technology will enable a broader patient access to biopharmaceuticals.
- Aiming to create new and breakthrough technologies, the action will boost competitiveness, job creation and growth, as well as build more efficient and cost effective DSP. In this way nextBiopharmDSP will help to achieve the EU’s Industrial policy’s goals. The advances for the EU’s global competitiveness are clear: the reduction in the amount of required initial investment and in running costs of the process will promote fast diffusion of the technology. The companies involved will have resources to reinvest in further R&D and in the short term create new jobs and growth.
- Lastly our technology contributes to a more competitive EU manufacturing sector, providing a technological solution that will help the European Union face its societal challenges in health and climate change, considered its beneficial impacts on resource consumption and pollutants reduction.

The project results should eventually lead to:
- Broader patients’ accessibility to highly efficient biopharmaceuticals;
- Smaller Health Care System burden;
- Highly efficient as well as more sustainable production processes;
- Reduced utilisation of materials (chromatography resins, buffers, water) will substantially decrease the environmental footprint of the proposed process;
- Further environmental benefits: large decrease in water consumption and reduction of relative CO2 emissions;
- Sizable reduction of required investments and operating costs (smaller buildings, lower facility footprint - reflected in reduced running costs and decreased energy consumption);
- Increased production flexibility when changing between different products;
handout nextBioPharmDSP
scheme of the proposed vs. current process