Increasing productivity of IVT reaction using at-line monitoring by HPLC chromatography
Apr
12
2022
On demand

Increasing productivity of IVT reaction using at-line monitoring by HPLC chromatography

Tuesday 08:00 PDT / 11:00 EDT / 16:00 BST / 17:00 CEST
Sponsor
Increasing productivity of IVT reaction using at-line monitoring by HPLC chromatography

mRNA is produced by in vitro transcription (IVT) reaction, which is typically performed as a batch process. Considering its catalytic basis, it is possible to extend reaction time and yields by continuous addition of consumed reagents to the reaction mixture, however fed-batch strategies reported to date have only managed to achieve a 40 – 100 % increase in the production of mRNA. One of the main limitations of development has been low throughput of analytics for quantification of mRNA and NTP precursors; productivity is typically determined as an end-point measurement of mRNA concentration.

Webinar will discuss an HPLC-based analytical methodology 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.

When applied to monitoring IVT reaction a batch approach with an average productivity of 3-5 mg/mL can be converted to fed-batch yielding 10-12 mg/mL. Two approaches for controlling IVT reaction will be demonstrated, one focusing on productivity of uncapped mRNA, another on productivity of capped mRNA, thereby requiring a precise control of NPT:capping reagent ratio

Attend this webinar to increase your understanding of:

  • Benefits of implementing HPLC-based analytics in mRNA production workflow
  • Principles of monolith chromatography and its use for mRNA analytics and purification
  • Principles of IVT reaction and factors affecting rate and yield
  • Principles of conversion from batch to fed-batch process
Rok Sekirnik
Rok Sekirnik
Head Process Development for mRNA and pDNA at BIA Separations
Rok Sekirnik graduated with a degree in chemistry from the University of Oxford, where he also completed his PhD. He has worked in the areas of mAb, vaccine and gene therapy vector development in various roles, including Principal Scientist at Novartis and Project Manager at Batavia Biosciences. His team at BIA Separations is focused on developing high-yielding, cost-effective purification approaches for pDNA and mRNA production.

SPEAKERS

Rok Sekirnik
Rok Sekirnik
Head Process Development for mRNA and pDNA at BIA Separations

You might also like

Same-section spatial multiomics: a platform for detailed analysis of the solid tumor TME

Same-section spatial multiomics: a platform for detailed analysis of the solid tumor TME

Emily Neil, Ariadna Pascual
27 June
in 70
Days
Register
Pure and simple: understanding LNP analytics for better mRNA-based drugs

Pure and simple: understanding LNP analytics for better mRNA-based drugs

Adam Crowe
29 May
in 41
Days
Register
Empowering patient care: in-process and quality control solutions for cellular therapies

Empowering patient care: in-process and quality control solutions for cellular therapies

Christin Tischner, Simon Lennartz
23 May
in 35
Days
Register
T cells from beginning to end: optimizing leukopak and drug product cryopreservation

T cells from beginning to end: optimizing leukopak and drug product cryopreservation

Meredith Safford, ShaNelle Yelity
22 May
in 34
Days
Register
Mastering AAV production: best practices For small- to large-scale manufacturing success and reduced development time

Mastering AAV production: best practices For small- to large-scale manufacturing success and reduced development time

Emily Jackson-Holmes
21 May
in 33
Days
Register