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@ARTICLE{Wohlfarth:177493,
author = {D. Wohlfarth$^*$ and V. Frehtman$^*$ and M. Müller$^*$ and
M. Vogel$^*$ and L. M. P. Phan$^*$ and A. Brunecker$^*$ and
B. Leuchs$^*$},
title = {{U}pstream process optimization and micro- and macrocarrier
screening for large-scale production of the oncolytic {H}-1
protoparvovirus.},
journal = {Applied microbiology and biotechnology},
volume = {105},
number = {24},
issn = {1432-0614},
address = {New York},
publisher = {Springer},
reportid = {DKFZ-2021-02580},
pages = {9113-9124},
year = {2021},
note = {#EA:F010#LA:F010# / 2021 Dec;105(24):9113-9124},
abstract = {The oncolytic virus H-1PV is a promising candidate for
various cancer treatments. Therefore, production process
needs to be optimized and scaled up for future market
release. Currently, the virus is produced with minimum
essential medium in 10-layer CellSTACK® chambers with
limited scalability, requiring a minimum seeding density of
7.9E3 cells/cm2. Production also requires a $5\%$ fetal
bovine serum (FBS) supplementation and has a virus yield up
to 3.1E7 plaque-forming units (PFU)/cm2. Using the
animal-free cell culture medium VP-SFM™ and a new feeding
strategy, we demonstrate a yield boost by a mean of 0.3 log
while reducing seeding density to 5.0E3 cells/cm2 and
cutting FBS supplementation by up to $40\%$ during the
production process. Additionally, FBS is completely removed
at the time of harvest. Eleven commercial micro- and
macrocarriers were screened regarding cell growth,
bead-to-bead transfer capability, and virus yield. We
present a proof-of-concept study for producing H-1PV on a
large scale with the microcarrier Cytodex® 1 in suspension
and a macrocarrier for a fixed-bed iCELLis® bioreactor. A
carrier-based H-1PV production process combined with an
optimized cell culture medium and feeding strategy can
facilitate future upscaling to industrial-scale production.
KEY POINTS: • Virus yield increase and FBS-free harvest
after switching to cell culture medium VP-SFM™. • We
screened carriers for cell growth, bead-to-bead transfer
capability, and H-1PV yield. • High virus yield is
achieved with Cytodex® 1 and macrocarrier for iCellis® in
Erlenmeyer flasks.},
keywords = {Bead-to-bead transfer (Other) / Macrocarrier (Other) /
Microcarrier (Other) / Protoparvovirus H-1PV production
(Other) / Scale-up (Other) / Serum-free (Other)},
cin = {F010},
ddc = {570},
cid = {I:(DE-He78)F010-20160331},
pnm = {316 - Infektionen, Entzündung und Krebs (POF4-316)},
pid = {G:(DE-HGF)POF4-316},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:34782922},
doi = {10.1007/s00253-021-11642-y},
url = {https://inrepo02.dkfz.de/record/177493},
}