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High yield mRNA production process from E.Coli to highly pure mRNA
 
At this year's Cell & Gene Therapy Bioprocessing & Commercialization conference our CEO, Aleš Štrancar, presented a new purification approach starting from E.Coli all through to mRNA production. This new approach integrates a pDNA linearisation step before polishing (removal of linear and oc isoforms) of pDNA. The polishing step, placed after enzymatic linearisation, purifies linear pDNA from enzyme and other unwanted process impurities. The linearised pDNA is then used in IVT for production of mRNA. This approach further introduces mRNA purification tools for improved contaminant removal. Read more...
Viral Vector Purification
A Discussion of Current Challenges and Methods

Adeno-associated viral (AAV) vectors have become synonymous with gene therapy delivery. However, because they are produced in such small quantities and because their upstream processes carry comparatively large amounts of host-cell DNA and other impurities, AAV purification can be challenging. This eBook features a discussion among several industry experts (Aleš Štrancar, J. Michael Hatfield, and Pete Gagnon) and explores challenges specific to AAV purification, shedding light on whether current strategies and separation technologies are up to the task.
Challenges in Industrial Process Development of Exosome-Based Therapies: Characterizing and Managing Diversity

The traditional classification of extracellular vesicles (EVs) includes three types: exosomes, microvesicles, and apoptotic vesicles. Each type arises from a distinct origin and exhibits distinct characteristics, but their size ranges overlap and the major surface proteins presented by exosomes also are present on the surfaces of microvesicles and apoptotic bodies. This makes it a challenge for process developers to identify the vesicle fraction that best serves a particular exosome therapy. This article highlights the complementarity of two analytical methods for characterizing distinctions among EV populations separated by anion exchange chromatography: imaging flow cytometry (IFCM) and size-exclusion chromatography.
One-Step Purification of Research-Grade Single-Stranded mRNA

CIMac PrimaS™ is a new member of BIA’s family of high performance monoliths for analysis and purification of mRNA. Its positive charge gives it some anion exchange behavior but hydrogen bonding makes its selectivity entirely distinct from traditional AEX.

QA and DEAE anion exchangers need to be heated into the range of 50–70°C for large mRNA to elute. PrimaS elutes mRNA in a pH gradient, well separated from DNA and dsRNA.
This Technical note presents new experimental data showing that NaCl shifts the elution profile to lower pH values.
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