Seminars & Webinars
2023
Event: mRNA Applications in Discovery & Development
Date: June 20, 2023
Presenter: Rok Sekirnik, Sartorius BIA Separations
This talk presents case studies of leveraging the power of at-line HPLC monitoring for process development to increase purity and yield of mRNA production. It describes approaches to optimize plasmid purification and linearization, IVT reaction and mRNA purification, including purification methods to achieve more than 95% supercoiled plasmid purity, optimize linearization reaction, and increase of IVT reaction yield from 3-5 mg/mL to 10-12 mg/mL. Further, use of at-line analytics to monitor automated, continuous production in single use bioreactors to produce multi-gram amounts of mRNA are demonstrated. Oligo dT-based affinity purification is described with approaches to optimize dynamic binding capacity to reach up to 6 g/L. Multimodal chromatography and reverse-phase chromatography cases are discussed for capture of non-polyadenylated RNA, and removal of dsRNA.
Event: Cell and Gene Therapy Insights webinar
Date: March 1, 2023
Presenter: Blaž Bakalar, Product Manager PATfix platform
mRNA-based therapeutics represent a promising new modality for therapies and vaccines. However, industry-wide process understanding of the CQA and CPP interplay is still developing. As such, the lack of immediate feedback on the process state during process optimization, sometimes delayed by weeks and limited by throughput, represents a big bottleneck in mRNA commercialization. The PATfix mRNA analytical platform enables reliable at-line insight during process development and production of mRNA-based therapeutics. With an ability to resolve key IVT reaction components (NTPs, capping reagent, template and mRNA), both USP and DSP process insight can be gained, with a single analytical tool. The PATfix mRNA platform has been upgraded to include LNP encapsulation analytics, enabling at-line monitoring of a crucial process step on the path from mRNA drug substance to LNP drug product.
Presenter: Blaž Bakalar, Product Manager PATfix platform
Date: February 15, 2023
Event: 2nd mRNA Analytical Development Summit 2023
Abstract:
- PATfix mRNA analytical platform is a perfect tool for process developers looking to optimize their mRNA production and purification
- Three orthogonal analytical methods based on mix-mode, affinity and reverse phase allow for precise quantification and mRNA sample characterization
- The newly developed method for characterization of encapsulated mRNA molecules covers the last crucial part in mRNA therapeutic production
Attachments
2022
Event: Cell and Gene Therapy Insights webinar
Date: June 7, 2022
Presenter: Nejc Pavlin
Pre-developed and validated PATfix analytical methods using different column chemistries enable:
- Effective control over linear plasmid upstream and downstream,
- IVT reaction optimization for mRNA synthesis,
- Control of yield, purity and downstream process of mRNA,
- mRNA formulation and stability analysis,
- Scale-up optimization.
In this webinar, will discuss a robust analytical tool PATfix™ platform, and its specific methods for pDNA and mRNA analytics that enable efficient monitoring of key upstream and downstream process steps will be showcased.
Event: LCGC Live Webcast
Date: March 17, 2022
Presenters:
Rok Zigon, Project Manager; Process Development Dept.
Blaz Goricar, Project Manager; Process Analytical Technology Dept.
Key learning points:
- Rapid downstream process of AAV using monoliths, with emphasis on PATfix analytical tools
- Orthogonal analytics are paramount for AAV characterization
- Integration using PATfix enables fast evaluation of empty-full ratio
Event: BPI Ask the Expert
Date: March 3, 2022
Speaker: Rok Sekirnik, Head Process Development for mRNA and pDNA
In this webinar, we will discuss an HPLC-based analytical method using a multimodal ligand based on anion exchange/hydrogen-bonding ligand (PrimaS) to monitor the IVT reaction, which allows for simultaneous quantification of NTPs, capping reagent, plasmid, and mRNA within 3.5 min.
Event: Cell and Gene Therapy Insights webinar
Date: February 17, 2022
Presenter: Ales Strancar, Managing Director
Key learning points:
- One of the key challenges in manufacturing viral vectors is to increase the ratio between empty and full capsids.
- When the expression in the cell line results in less than 10% full capsid it is quite impossible to reach better than 90% full in the final product. It is therefore mandatory to optimise the USP to result in better empty/full ratio.
- This can be efficiently realised by using at-line PATfix to allow for analysis of the full and empty capsids ratio directly in the harvest.
- The residual empty capsids can be removed by polishing step using different anion exchange columns.
2021
Published: Cell & Gene Therapy Insights 2021; 7(12), 1661
Date: December 20, 2021
Presenter: Maja Leskovec, Head of Process Development Viruses (viral vectors and vaccines)
Watch the video or read the poster to learn:
- Importance of sample pretreatment before first chromatographic step
- Rapid downstream process of AAV with emphasis on chromatographic step of CIMmultus SO3 capture
- Three different ligands full enrichment of AAV
- Why orthogonal analytics is the key in downstream development and what are the options
Attachments
Published: Cell & Gene Therapy Insights 2021; 7(11), 1409
Date: December 2, 2021
Presenter: Rok Sekirnik, Head Process development for mRNA and pDNA
Watch the video or read the poster to learn:
- Non-affinity chromatography with PrimaS provides purification for IVT processes with co-transcriptional polyadenylation
- The Sartorius mRNA purification toolbox provides a scalable chromatographic solution for commercial manufacturing of mRNA therapeutics
- CIMac analytical HPLC columns can resolve individual IVT reaction components in minutes, resolving the analytical bottleneck in optimization of IVT reaction conditions
- Rapid HPLC analytics can improve in-process and quality control of the mRNA production process
Attachments
Published: Cell & Gene Therapy Insights 2021; 7(10), 1437
Date: November 15, 2021
Presenter: Nejc Pavlin, Project Manager in Process Analytics Development department
Watch the video or read the poster to learn:
- The importance of pDNA as an enabling product in the production of mRNA, AAV, and other therapeutic vectors
- How the PATfix™ pDNA platform using CIMac™ pDNA column offers fast and efficient solution for problems related to pDNA upstream and downstream processes
- How the PATfix™ pDNA platform can be used to optimally adjust pDNA production and purification procedures of pDNA according to your needs, increasing pDNA yield and purity and reducing the cost of its production
Attachments
Presenter: Ivana Petrović Koshmak, PhD
Event: Late Stage Bioprocessing & Viral Vectors 2 11 November 2021
Date: November 3, 2021
Abstract:
Adeno associated virus (AAV) is the leading vector in the field of gene therapy because of its low toxicity, good overall safety profile, and ability to maintain stable expression for long periods of time. It is therefore crucial to develop a robust and high efficiency platform for its manufacturing.
One of the key challenges in manufacturing viral vectors is to increase the ratio between empty and full capsids. The most efficient way is to design the USP to result in less than 10% of empty capsids. This can be realised by process optimisation using at-line PATfix analytics to allow for analysis of the full and empty capsids ratio during the virus expression.
Fast and reliable at-line analytical methods to allow for process optimisation USP and DSP, and assessing the purity of the final product using PATfix system will be presented.
Attachments
Presenter: Aleš Štrancar
Event: BPI and Cell & Gene Therapy Bioprocessing & Commercialization 2021
Date: September 28, 2021
Abstract:
Adeno associated virus (AAV) is the leading vector in the field of gene therapy because of its low toxicity, good overall safety profile, and ability to maintain stable expression for long periods of time. It is therefore crucial to develop a robust and high efficiency platform for its manufacturing.
One of the key challenges in manufacturing viral vectors is to increase the ratio between empty and full capsids. The most efficient way is to design the USP to result in less than 10% of empty capsids. This can be realised by process optimisation using at-line biochromatography analytics to allow for analysis of the full and empty capsids ratio directly in the harvest.
The residual empty capsids can be further removed by polishing purification step using different anion exchange columns.
Fast and reliable at-line analytical methods to allow for process optimisation and assessing the purity of the final product using PATfix system will be presented.
Attachments
Presenter: Aleš Štrancar
Event: BPI and Cell & Gene Therapy Bioprocessing & Commercialization 2021
Date: September 30, 2021
Abstract:
Global demand for pDNA production is at an all time high, due to increased need from Gene Therapy ramp-up. pDNA, as an enabling product, is critical in production of mRNA, AAV and other therapeutic vectors. Increasing yield and purity in the production of pDNA is a vital step in meeting such demand. Supporting reliable in-process control during pDNA purification, PATfix pDNA analytical platform is enabling rapid process development and optimization while providing a reliable analytical platform for production runs.
Attachments
Presenter: Ivana Petrović Koshmak
Event: 2nd Annual Gene Therapy Analytical Development EU
Date: May 26, 2021
Abstract:
In-process analytics of vector capsid production is a critical optimization target in development of AAV-based gene therapy products. In this presentation we introduce a fast at-line HPLC based system that enables differentiation of vector capsid and empty capsid production in transfection mixtures. Case study results demonstrating the effects of different process variables on capsid production and assembly are shown and insights about purification are also discussed.
Attachments
2020
Messenger RNA is poised to become a major contributor in the fields of gene therapy and vaccines. Making this a practical reality requires purification technology that accommodates its unique features and challenges. Some of those challenges are inherent to mRNA. Others are byproducts of RNA synthesis.
This presentation addresses both and introduces a coordinated purification toolbox to advance the evolution of mRNA therapy.
Attachments
Presenter: Ingo Nagler
Date: June 3, 2020
Biomanufacturing and Cell & Gene Strategy Meetings (Proventa International's online meetings)
Click here to listen to BIA's Keynote Presentation
Attachments
Presenter: Ingo Nagler
Date: May 14, 2020
Asia’s SARS-CoV-2 Vaccine Development Challenges (Corona360 In-Conversation webcast series)