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Seminars & Webinars

2022

Event: Informa 1-day digital event, highlighting mRNA Applications and Therapeutics
Date: March 23, 2022
Presenter: Tomas Kostelec, Technology & Business Development Manager

Key learning points:

  • Ensuring consistent and robust IVT production of mRNA 
  • Achieving high yield and high purity in mRNA purification processes 
  • Chromatography solutions for in process analytics and purification

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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 HPLC analytical tools
  • Orthogonal analytics are paramount for AAV characterization
  • Integration using PATfix enables fast evaluation of empty-full ratio

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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 HPLC 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.

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Event: mRNA-Based Therapeutics Summit Europe
Date: January 2022
Presenter: Rok Sekirnik, Head of Process development for mRNA and pDNA

Abstract:

In vitro-transcribed messenger RNA-based therapeutics represent a relatively novel and highly efficient class of drugs. Current clinical efforts encompassing mRNA – based drugs are directed to three categories: mRNA vaccine, mRNA therapy and protein replacement therapy. As mRNA vaccine became the front runner to fight the COVID-19 pandemic, challenges surrounding their process development and manufacturing became readily apparent:

  1. mRNA manufacturing is not yet a platform approach. Standardized products along the processes are needed
  2. mRNA is a relatively large molecule, which leads more challenges for downstream purification.
  3. mRNA is intrinsically unstable and prone to degradation.
  4. High raw material cost is limiting mRNA commercialization.

This presentation provides a detailed overview of current mRNA manufacturing approaches, highlights challenges and recent successes, and offers perspectives on the future of mRNA manufacturing.

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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

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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

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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

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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 HPLC to allow for analysis of the full and empty capsids ratio during the virus expression.

Fast and reliable in HPLC methods to allow for process optimisation USP and DSP, and assessing the purity of the final product using PATfix system will be presented.

Attachments

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Published: Cell & Gene Therapy Insights 2021; 7(8), 1125
Date: October 4, 2021
Presenter: Hana Jug, Project Manager in process development for viral vectors and vaccines

Watch the video or read the poster to learn: 

  • Why downstream processing remains one of the main bottlenecks in adenoviral vector manufacturing 
  • How an adenoviral vector purification platform using CIMmultus™ QA as the key purification step, secures a fast and robust process with better purity 
  • CIMmultus™ QA offers high capacity and high yields of adenovirus, results in cost reduction and overcomes raw material supply bottlenecks

Attachments

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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

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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 HPLC 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 in HPLC methods to allow for process optimisation and assessing the purity of the final product using PATfix system will be presented.

Attachments

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Presenter: Rok Sekirnik
Event: mRNA Therapeutics Summit
Date: July 14, 2021

Abstract:

The recently demonstrated efficacy of mRNA-based Covid-19 vaccines has shown the promise of this therapeutic format, but also highlighted the need for higher efficiency of mRNA production to meet enormous needs for global vaccine supply. Typical mRNA production process involves three key steps: 1) plasmid DNA (pDNA) production in supercoiled (sc) isoform, linearization and purification, 2) in-vitro transcription (IVT) reaction and 3) mRNA purification. Here we present a chromatographic toolbox for integrated mRNA production from pDNA to mRNA purification, including in process analytics.

Attachments

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Presenter: Aleš Štrancar
Event: TIDES mRNA Digital Week
Date: June 28, 2021

Abstract:

A new purification approach starting from E.Coli all through to mRNA production is presented here. This new approach integrates a pDNA linearisation step before polishing (removal of linear and open circular isoforms) of plasmid DNA. The polishing step, placed after enzymatic linearisation, purifies linear pDNA from enzyme and other unwanted process impurities. The linearised plasmid DNA is then used in IVT for production of mRNA. Fast in-process analytics allows for transcription number well above 100 in reproducible manner.

This approach further introduces mRNA purification tools for improved contaminant removal. The altered sequence of purification and linearisation reduces the overall number of purification steps required, improves recoveries, while the complete process results in extra low protein impurity and very efficient dsRNA removal.

The participants would get insight into the mRNA production steps, their optimisation and tools to allow for robust process. The analytical tools to allow for safe product will be highlighted, too.

Attachments

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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

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Presenter: Aleš Štrancar
Event: Genetic Vaccine Development for Infectious Diseases Summit
Date: May 20, 2021

Abstract:

  • Developments in gene therapy and threat of pandemics requests novel technologies to allow for efficient manufacturing of very large biomolecules and nanoparticles within short time.
  • Convective Interaction Media with open channel structures resulting in no shear forces allows for fast and very efficient manufacturing of mRNA, pDNA, Adeno and other VLP based Covid vaccines.
  • Ultra-sensitive DNA assay and fast in-process control using PATfix systems are introduced for production of safer Covid vaccines.

Attachments

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Presenter: Aleš Štrancar
Event: ASGCT 24th Annual Meeting
Date: May 13, 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. Therapeutic applications of AAV-based gene therapy vectors require the process and product related impurities to be removed, as they represent serious safety threats as well as burden the economics of manufacturing.

Attachments

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Event: BioProcess International COVID-19 Therapeutic Development & Production
Date: April 27, 2021

Developments in gene therapy and the threat of pandemics requires novel technologies to allow for efficient manufacturing of exceptionally large biomolecules and nanoparticles within a short time. Novel vaccine platforms such as mRNA enable the industry to meet its highest goals: decreasing footprint while increasing efficiency and throughput. However, to respond with agility and prepare for future outbreaks, the mRNA platform must be further optimized.

In vitro transcription (IVT), the enzymatic process used for mRNA vaccine production, is different from biological fermentation processes as it requires linearized plasmid DNA. The linear isoform is produced with restriction enzymes from open circular and supercoiled pDNA. Employing a traditional pDNA manufacturing process, which removes multimers as well as linear and open-circular isoforms, reduces production yield.

However, when pDNA and mRNA are treated as a single production process, the purification steps are optimized, yields improved, and overall production costs reduced. This presentation will describe how Convective Interaction Media monoliths can be used to efficiently purify a board specter of mRNA and cover all production scenarios.

This presentation illustrates fast in-process control methods using the PATfix system, which can ensure the rapid production of the safest possible Covid-19 vaccines while reducing manufacturing costs.

Attachments

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Event: Nucleic Acids- cGMP Production of mRNA & pDNA (virtual edition)

Date: February 1, 2021

Our new Head of Process Development for mRNA and pDNA purification, Rok Sekirnik, presented our Chromatographic tools for high-yielding mRNA production process as workshop.

Click here to listen to the workshop.


Event: Nucleic Acids- cGMP Production of mRNA & pDNA (virtual edition)

Date: February 3, 2021

Our CEO, Ales Strancar presented our Integrated Production Process of mRNA from E. coli to Highly Purified mRNA

Click here to listen to the presentation.

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2020

Presenter: Aleš Štrancar

Date: October 27, 2020

Gene Therapy for Rare Disorders Europe 2020 (digital event)

Bullet points:

  • Buffer optimisation and proper column ligand selection (CIM QA, CIM PrimaS or new CIM EF) allows for >90% full capsid production
  • Introduction of Specimen allows for production columns checking before use  
  • Using ultracentrifuge with set of HPLC detectors allows for prompt orthogonal verification of the full capsid purity  

Attachments

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Presenter: Aleš Štrancar

Date: od demand, October 19-22, 2020

Cell & Gene Therapy Bioprocessing & Commercialization  (digital event)

Established processes for pDNA manufacturing include purification steps designed to enrich the supercoiled (sc) isoform. Homogeneous supercoiled isoform improves cell transfection efficiency in fermentation processes and is favoured in cell culture production systems. In vitro transcription (IVT), the enzymatic process used for production of mRNA vaccines differentiates itself from biological fermentation processes by the need for linearised plasmid DNA. The linear isoform is produced with restriction enzymes from open circular and supercoiled plasmid DNA. Employing a traditional pDNA manufacturing process, which removes linear and open-circular isoforms, will therefore reduce the production yield. When plasmid DNA and mRNA are treated as a single production process, the purification steps can be optimised, yields improved, and the overall production cost reduced.

A new purification approach starting from E.Coli all through to mRNA production is presented here. This new approach integrates a pDNA linearisation step before polishing of plasmid DNA. The polishing step, placed after enzymatic linearisation, purifies linear pDNA from enzyme and other unwanted process impurities. The linearised plasmid DNA is then used in IVT for production of mRNA. This approach further introduces mRNA purification tools for improved contaminant removal. The altered sequence of purification and linearisation reduces the overall number of purification steps required, improves recoveries, while the complete process results in extra low protein impurity and very efficient dsRNA removal.

Attachments

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