GEN-Thermo_Oct22_MakingThePromiseOf - 21

OVERCOMING mRNA PRODUCTION SCALE-UP CHALLENGES
Question 4: Which part of IVT scale-up requires the most optimization?
" The balance between mRNA yield and quality. "
-Pirkko
" Assessing different mixing speeds at an intermediate
scale before scaling up. Large volumes of raw
material will require different mixing to ensure a
homogeneous solution. " -Sirat
" The most optimization may be required to obtain
the highest yield in the system the customer is
using-to select reaction volume, duration of reaction,
whether additional supplementation of NTPs,
enzymes, or other components is needed during
incubation, stability of the buffer (e.g., considering
DTT or other reducing agents), or shaking
during the IVT reaction (e.g., shaking speed). In
my opinion, every customer needs to adjust these
parameters in their system to reach the desired
amounts of RNA synthesized in the shortest time
as possible, because longer incubation times may
also affect RNA integrity, especially if the transcript
is long (e.g., self-amplifying RNA). " -Monika
Question 5: What questions are you most commonly asked?
" What vessels should I use in different scales? My
answer is moving from tubes to shake flasks at
around 15-20 mL. It's then beneficial to start using
controlled vessels, like bioreactors, when producing
GMP materials. " -Pirkko
" Is the IVT yield or integrity impacted as production
is scaled up? It may be, which is why it is important
to understand the impact of all process parameters
on process performance and critical quality attributes
of the target product. " -Sirat
" Does Thermo Fisher have GMP-grade raw materials
for IVT that I can use for clinical trial manufacture?
Yes, we have the TheraPure GMP product
portfolio with almost all critical enzymes and NTPs,
which can be provided with necessary documentation.
We perform validation runs before
starting manufacturing for each raw material, to
help ensure process consistency and QC before
releasing the raw materials. Thermo Fisher can also
supply large quantities of the enzymes and NTPs
when you choose to scale up production. " -Sirat
Question 6: What are your most useful hints and tips for successful IVT scale-up?
" When testing low and high temperature conditions,
keep in mind that lower temperatures require
longer IVT reaction times. But higher temperature
conditions may be limited to 40-45°C, as single-use
plastics used during large-scale production do not
necessarily tolerate high temperatures. " -Pirkko
" Not all " GMP-grade " materials are the same in
terms of product quality and the quality systems
supporting those products. Request data on
raw material quality and lot-to-lot consistency
before selecting a supplier for scale-up, as these
can significantly impact product quality. OptiGENengnews.com
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