Bioprocessing Webinar Series●

Abstract:

Purification of monoclonal antibodies by Protein A resin is an industry standard operation but developing the process for efficient manufacturing scale production can be time intensive. This presentation will cover our historical approach to ProteinA process development followed by how we have used a Tecan with Robocolumns to improve that process yielding significant time and material savings without sacrificing the accuracy of our data.

Abstract:

Scaling down our chromatography processes has provided a useful, high-throughput and automated screening tool, that can help us accelerate timelines and save resource. We have been working on increasing our automation capabilities to try and accomplish a full end-to-end downstream process, that we can use for process development of our assets. Using OPUS RoboColumns on the Tecan EVO and Fluent workstations, we have developed various chromatography scrips to investigate the behaviours of our molecules in flow-through mode, with the ability to screen multiple condition in parallel. This has helped us better characterise our molecules, how they interact with different resins and optimise the separation of product from impurities.

Abstract:

This webinar describes automated high throughput parallel protein purification using Repligen OPUS® RoboColumns® on the Tecan Fluent® Automation Workstation – a market-leading solution. Benefit from the expert partnership between Tecan and Repligen, bringing together years of experience and joint application support.

Key themes:
- Increase throughput by processing up to eight OPUS RoboColumns in parallel
- Validate chromatographic conditions without the need for complex LC systems, for example, to predict manufacturing scale performance

From simple benchtop systems to complex automated solutions to run multiple assays.

Abstract:

Purification of adeno-associated vectors (AAV) remains a challenge and can easily create a bottleneck in the process development for the manufacturing of therapeutic genes. Join this free on-demand webinar and learn how to fully automate miniaturized column chromatography to speed-up your purification process development by 8-fold. In the second part, discover the various possibilities Repligen offers with OPUS® Pre-Packed Columns to streamline AAV purification.

Abstract:

Miniaturization, parallelization and lab automation are powerful approaches to increase cultivation throughput in microbial phenotyping, metabolic engineering and bioprocess development. However, to fully release the potential of such approaches, standard and tailor-made automated workflows need to be put in place, comprising the full experimental cultivation pipeline from upstream processing, batch and fed-batch cultivation processes, process analytics, data management and design-of-experiment. This is illustrated using case studies with microbial systems, demonstrating how developments in miniaturized cultivation combined with smart lab automation, analytics and data processing are amalgamated in workflows for more efficient microbial phenotyping and bioprocess development.

Abstract:

The development of serum-free (SF) and chemically defined media (CDM) for the clinical production of human Mesenchymal Stem Cells (MSC) is a high priority within the cell therapy community. Small molecules as media supplements are an attractive tool as they are generally easily available, chemically defined and tunable, and chemical media design presents an attractive opportunity for improving MSC manufacturing. In this context, we are developing a novel analytical technology aimed at designing chemically defined media (CDM) tailored to the MSC production process. For this purpose we have established an automated high-content imaging assay to characterize the morphological profile of hMSCs in response to different growth conditions. This assay, multiplexed with a viability assay, drives the high-throughput screening of a suits of small molecules on a serum free background, with the goal of identifying molecules that sustain proliferation of MSC while retaining their potency. Importantly, the assay may represents a straightforward strategy to benchmark MSC preparation and media formulations during production processes for therapeutic application.

Key focus area:
Cell Therapy, Media Development, Automation, High Throughput Screening.

Speaker Bio:
Marilina Piemontese holds a Masters degree in Biotechnology and a PhD in “Skeletal Biology and Molecular Pathology” from the University of Parma, Italy. During her career, Marilina acquired extensive knowledge in Mesenchymal Stem Cell (MSC) cell culture and biology. From 2012 to 2016 Marilina was a visiting research fellow at the University of Arkansas for Medical Sciences (UAMS, USA) where she successfully managed several preclinical projects in the field of skeletal biology. In 2016 Marilina joined the Telethon Institute for Genetics and Medicine (Italy) where she worked on a project aimed at identifying new inducers of autophagy by high-throughput screening of small molecules to treat lysosomal storage disorders. In 2019 Marilina was awarded the Irish Research Council Enterprise Postdoctoral Fellow to work with Prof. Frank Barry’s laboratory at REMEDI (NUIG, Ireland) on collaborative work with industry partner Valitacell.Her research interests include the development of analytical technologies for MSC manufacturing, with a specific focus on establishing a novel media design platform for Mesenchymal Stem Cell (MSC) manufacturing.

Abstract:

When considering the development of therapeutics, vaccines in particular, research and development work is labour intensive, often requires lengthy timescales and is costly to perform. High throughput screening methodologies can support and speed up process development and obtain significant amounts of data early to de-risk vaccine manufacture. When coupled with automation, these screening platforms can significantly reduce the operator burden during optimisation studies and reduce the variability attributed to manual handling.

For the development of an automated screening platform the inherent variability associated with biological cell culture is often accounted for by the end-user and some robotic manipulations must be manually adapted for each iteration of a new experimental run. The ability to create an integrated generic platform, capable of adaptation would greatly enhance the generation of a true ‘smart’ screening platform with ‘plug and play’ functionality.

In this work we present a platform which provides the end-user with the flexible operation of a process sequence on an automated liquid handling system. The functionality of this platform is demonstrated from the conversion of a benchtop vaccine production process to the microscale, such as to investigate various operating conditions during upstream development. The application is shown to allow hands-free operation from the point of preculture until the loading of a western blot for product analysis. The use of such a platform can immensely simplify development activities and is accessible for non-automation experts once established.

Abstract:

It is necessary to screen using purified antibody at all stages of antibody generation to obtain the most reliable assay results. We have designed and implemented an automated high throughput process for the rapid generation of tens of thousands of antibodies. We have also introduced an HTP Process for generating matrices of thousands of bispecific antibodies which is vital to the screening process.

Key focus area:
Bispecific antibody generation, high-throughput screening platform, high throughput purification, biologics drug discovery.

Speaker Bio:
As an experienced and innovative scientist Jason has developed a high-throughput automated antibody production platform at Pfizer. Jason has extensive experience in developing, implementing and automating processes to produce, screen and characterize antibodies & proteins for all Research Units across Pfizer’s portfolio. Jason is currently enrolled in an MBA Program at Babson College.

Abstract:

All too often, the translation from development to manufacture of new vaccines relies on protocols and methodologies that were sufficient to allow development of candidate products to proof of concept and clinical trial rather than being optimized for efficiency and cost. In addition, the development of upstream and downstream processing are often treated as independent units. The combination of these factors can result in inefficient production systems with increased production costs. To prevent this from happening scalable manufacturing processes and appropriate analytics need to be defined early in the development process to speed manufacturing development.

We have taken a holistic approach to the development of a new scalable manufacturing process for a subunit rotavirus vaccine candidate, thereby speeding and simplifying the development process.

To accomplish this we made use of automated scale-down systems for screening and development of both upstream fermentation and downstream chromatography.

This whole process approach allowed identification of compatible upstream and downstream conditions, where product quality was improved in fermentation and maintained throughout the purification process. This entire process is scalable and compatible with an integrated, automated, modular manufacturing platform.

Abstract:

In collaboration with Tecan, Synthace Ltd has developed an integrated, end-to-end protocol on using their software Antha that cuts down time needed to plan, track, execute, analyse and share miniaturized purification workflows, thereby facilitating high throughput process development (HTPD) and process analytical technology (PAT).

In this presentation, we firstly demonstrate the flexible design of a purification workflow, and the data alignment capabilities, yielding a 5-fold decrease in time to visualisation. Next, we show how A280 absorption data can easily be augmented with external data, for example from a LabChip® GXII electrophoresis system. Finally, we present how instructions for a 24-parameter, 6-resin DOE are effortlessly created, split across independent runs, and how data from these runs is effortlessly combined together.

Key focus areas:
High throughput purification, laboratory data management, Design of Experiments.

Speakers Bio:
Dr Arne Vandenbroucke is the Product Lead for Miniaturised Purification and Senior Automation Engineer at Synthace Ltd. He leads the product strategy and development for Synthace’s partnership with Tecan to integrate the Antha software platform with Tecan’s Te-Chrom™ automated chromatography systems on Freedom EVO® and Fluent® platforms. Prior to joining Synthace, Dr Vandenbroucke developed liquid handling and molecular imaging systems, automated workflows, and data analysis pipelines at Labcyte Inc., Specific Diagnostics LLC, and Stanford University. He completed his PhD in Experimental Particle Physics at Ghent University.