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000177493 0247_ $$2doi$$a10.1007/s00253-021-11642-y
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000177493 0247_ $$2ISSN$$a0171-1741
000177493 0247_ $$2ISSN$$a0175-7598
000177493 0247_ $$2ISSN$$a0340-2118
000177493 0247_ $$2ISSN$$a1432-0614
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000177493 037__ $$aDKFZ-2021-02580
000177493 041__ $$aEnglish
000177493 082__ $$a570
000177493 1001_ $$0P:(DE-He78)b3beb23586d3d89554eeb27e1c384463$$aWohlfarth, Daniel$$b0$$eFirst author$$udkfz
000177493 245__ $$aUpstream process optimization and micro- and macrocarrier screening for large-scale production of the oncolytic H-1 protoparvovirus.
000177493 260__ $$aNew York$$bSpringer$$c2021
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000177493 500__ $$a#EA:F010#LA:F010# / 2021 Dec;105(24):9113-9124
000177493 520__ $$aThe 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.
000177493 536__ $$0G:(DE-HGF)POF4-316$$a316 - Infektionen, Entzündung und Krebs (POF4-316)$$cPOF4-316$$fPOF IV$$x0
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000177493 650_7 $$2Other$$aBead-to-bead transfer
000177493 650_7 $$2Other$$aMacrocarrier
000177493 650_7 $$2Other$$aMicrocarrier
000177493 650_7 $$2Other$$aProtoparvovirus H-1PV production
000177493 650_7 $$2Other$$aScale-up
000177493 650_7 $$2Other$$aSerum-free
000177493 7001_ $$0P:(DE-He78)0ee3163b1149540612c1397430e616d5$$aFrehtman, Veronika$$b1$$udkfz
000177493 7001_ $$0P:(DE-He78)e98381f0ca8b5d8503c582337efa5453$$aMüller, Marcus$$b2$$udkfz
000177493 7001_ $$0P:(DE-HGF)0$$aVogel, Martin$$b3
000177493 7001_ $$0P:(DE-HGF)0$$aPhan, Linh Minh Phuc$$b4
000177493 7001_ $$0P:(DE-He78)3eeba3804d05055920104ab28a2f56d7$$aBrunecker, Adrian$$b5$$udkfz
000177493 7001_ $$0P:(DE-He78)119d7d540a0e6fc5734898bef4866d7f$$aLeuchs, Barbara$$b6$$eLast author$$udkfz
000177493 773__ $$0PERI:(DE-600)1464336-4$$a10.1007/s00253-021-11642-y$$n24$$p9113-9124$$tApplied microbiology and biotechnology$$v105$$x1432-0614$$y2021
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