The Biopharmaceutical Production & Development Unit (BP&DU)
Barbara Leuchs
The Biopharmaceutical Production & Development Unit (BP&DU) supports research-associated topics
Viral vectors and oncolytic viruses
- Large Scale production and purification of parvovirus (vector) stocks for pre-clinical applications or basic research
- Development and optimization of new production, purification and analytic methods with the aim to establish working protocols for GMP-production of virus (vector) stocks necessary for clinical applications and possible market release.
- Accompanying research in two ParvOryx clinical trials with patient tumor and shedding assessment (see: Research - Treat (dkfz.de))
- In the establishment of GMP-produced parvovirus, the unit plays a pivotal role in the interaction and support of various organizations involved in the approval and application of procedures for virus (vector) production under GMP and GLP-conditions. Cooperation with GMP company (IDT Biologika GmbH) and GLP companies (Biogenes GmbH, Eurofins GmbH, Labor Enders & Kollegen)
Bovine Milk and Meat factors (in cooperation with Episomal-Persistent DNA in Cancer- and Chronic Diseases)
- Standardized production and purification of different BMMF´s in prokaryotic and eukaryotic cells.
- Characterization of BMMF´s regarding quantity, purity, aggregation, stability and activity, isoelectric point (pI), formulation
- Optimization of the process (upscaling, purity)
- Optimization of diagnostic assays in different matrices
- BMMF detection in animals and dietary products
- Stability and Stress tests of BMMFs in different formulations
Parasites
- Large scale (up to 5 Liter) production and purification of parasites including optimization strategies
DNA
- Large scale (up to 5 Liter) production and purification of pre- GMP- DNA
Assay-Development, Optimization and Qualification
- Divers immunological, biological and molecular-biological assays (e.g. ELISA, potency, qPCR, endotoxin, stability, deactivation) with standardized SOPs
- Stability and stress tests for possible drug candidates or antigens for diagnostic
- Consulting and technical support for production-related inquiries
Focus of the group lies on
- Upstream process: establishment, optimization and upscaling of production process
- Downstream process: optimization of purification process according to impurity profile, quality and quantity
- Qualified and validated analysis (quantity and quality)
- Establishment of good process and quality controls
- Safety assessment (i.e. establishment of optimal deactivation protocols)
- Consideration of ecological and economical aspects
- Statistical experiment planning using Design of Experiment approach
Methods-Portfolio
Upstream production process
- Micro- and macrocarrier up to 2.5 Liters for adherent cells
- Suspension (shaked or stirred) Bacteria and Parasites cultures up to 5 L
- Production in 10-layer Cell factories (6360 cm², Corning) or fixed bed bioreactor (4 m²) iCellis nano (Pall) For detailed description see Leuchs et al 2016 and Wohlfarth et al 2021
Downstream production process
- Clarification (centrifugation, filtration with diverse filters up to 0.2µm)
- Chromatographic purification with ÄKTApure (Cytiva). For H-1PV purification see Leuchs et al 2017
- Tangential flow filtration for separation, buffer exchange and concentration
- Sterile filtration and final formulation
Specific qualitative and quantitative Analysis
- Metabolite analysis (Glucose, Lactate, Ammonia, LDH, etc.) with CEDEX Bio analyzer (Roche)
- Cell and cell cycle characterization with Nucleocounter (Chemometec)
- Photometric, fluorometric and luminometric analytical methods with FLUOstar Omega (BMG Labtech) for e.g. ELISA, immunoassays, protein-, DNA analysis
- Characterization of the product using size exclusion chromatography, 2D electrophoresis, protein profile (WB), endotoxin assessment
- Stability testing as well as establishment of optimal deactivation protocol of H-1PV in regard to regulatory requirements (Frehtman et al; 2023)
Cooperations
Episomal-Persistent DNA in Cancer- and Chronic Diseases
Virotherapy (Clinical trials)
Training opportunities
6 Month training or bachelor or master theses for students from life sciences faculties or from technical schools are possible.
You are welcome to contact Barbara Leuchs (Engineer of Biotechnology)
Our team
5 Employees
Virus production, purification and specification related publications
Stability and safety key factors of the oncolytic protoparvovirus H-1 from manufacturing to human application
Veronika Frehtman, Daniel Wohlfarth, Marcus Müller, Ottheinz Krebs, Barbara Leuchs. Appl Microbiol Biotechnol. 2023 Aug;107(15):4777-4787
Upstream process optimization and micro- and macrocarrier screening for large-scale production of the oncolytic H-1 protoparvovirus
Daniel Wohlfarth 1, Veronika Frehtman 1, Marcus Müller 1, Martin Vogel 1, Linh Minh Phuc Phan 1, Adrian Brunecker 1, Barbara Leuchs 2
Appl Microbiol Biotechnol. 2021 Dec;105(24):9113-9124; doi: 10.1007/s00253-021-11642-y. Epub 2021 Nov 16.
Protocol for Efficient Generation and Characterization of Adeno-Associated Viral Vectors.
Jungmann A, Leuchs B, Rommelaere J, Katus HA, Müller OJ.
Hum Gene Ther Methods. 2017 Oct;28(5):235-246. doi: 10.1089/hgtb.2017.192. PMID:29048971
A novel scalable, robust downstream process for oncolytic rat parvovirus: isoelectric point-based elimination of empty particles.
Leuchs B, Frehtman V, Riese M, Müller M, Rommelaere J.
Appl Microbiol Biotechnol. 2017 Apr;101(8):3143-3152. doi: 10.1007/s00253-016-8071-x. Epub 2017 Jan 14.
Standardized large-scale H-1PV production process with efficient quality and quantity monitoring.
Leuchs B, Roscher M, Müller M, Kürschner K, Rommelaere J.
J Virol Methods. 2016 Mar;229:48-59. doi: 10.1016/j.jviromet.2015.11.022. Epub 2015 Dec 2.
Preclinical and clinical trail related publications
H-1 Parvovirus-Induced Oncolysis and Tumor Microenvironment Immune Modulation in a Novel Heterotypic Spheroid Model of Cutaneous T-Cell Lymphoma
Assia Angelova, Milena Barf, Alexandra Just, Barbara Leuchs, Jean Rommelaere, Guy Ungerechts
Cancers. 2024 Jul 30;16(15):2711
Phase 2 Trial of Oncolytic H-1 Parvovirus Therapy Shows Safety and Signs of Immune System Activation in Patients With Metastatic Pancreatic Ductal Adenocarcinoma.
Hajda J, Leuchs B, Angelova AL, Frehtman V, Rommelaere J, Mertens M, Pilz M, Kieser M, Krebs O, Dahm M, Huber B, Engeland CE, Mavratzas A, Hohmann N, Schreiber J, Jäger D, Halama N, Sedlaczek O, Gaida MM, Daniel V, Springfeld C, Ungerechts G.
Clin Cancer Res. 2021 Oct 15;27(20):5546-5556. doi: 10.1158/1078-0432.CCR-21-1020. Epub 2021 Aug 23. PMID: 34426438
Human Retrotransposons and the Global Shutdown of Homeostatic Innate Immunity by Oncolytic Parvovirus H-1PV in Pancreatic Cancer.
Neulinger-Muñoz M, Schaack D, Grekova SP, Bauer AS, Giese T, Salg GA, Espinet E, Leuchs B, Heller A, Nüesch JPF, Schenk M, Volkmar M, Giese NA.
Viruses. 2021 May 28;13(6):1019. doi: 10.3390/v13061019. PMID: 34071585
Fluorescence In Situ Hybridization (FISH) Detection of Viral Nucleic Acids in Oncolytic H-1 Parvovirus-Treated Human Brain Tumors.
Kiprianova I, Just A, Leuchs B, Rommelaere J, Angelova AL.
Methods Mol Biol. 2020; 2058:295-306. doi: 10.1007/978-1-4939-9794-7_20. PMID: 31486047
Oncolytic H-1 Parvovirus Shows Safety and Signs of Immunogenic Activity in a First Phase I/IIa Glioblastoma Trial.
Geletneky K, Hajda J, Angelova AL, Leuchs B, Capper D, Bartsch AJ, Neumann JO, Schöning T, Hüsing J, Beelte B, Kiprianova I, Roscher M, Bhat R, von Deimling A, Brück W, Just A, Frehtman V, Löbhard S, Terletskaia-Ladwig E, Fry J, Jochims K, Daniel V, Krebs O, Dahm M, Huber B, Unterberg A, Rommelaere
J. Mol Ther. 2017 Aug 24. pii: S1525-0016(17)30378-7. doi: 10.1016/j.ymthe.2017.08.016. [Epub ahead of print] PMID:28967558
A non-controlled, single arm, open label, phase II study of intravenous and intratumoral administration of ParvOryx in patients with metastatic, inoperable pancreatic cancer: ParvOryx02 protocol.
Hajda J, Lehmann M, Krebs O, Kieser M, Geletneky K, Jäger D, Dahm M, Huber B, Schöning T, Sedlaczek O, Stenzinger A, Halama N, Daniel V, Leuchs B, Angelova A, Rommelaere J, Engeland CE, Springfeld C, Ungerechts G.
BMC Cancer. 2017 Aug 29;17(1):576. doi: 10.1186/s12885-017-3604-y. PMID:28851316
Moving oncolytic viruses into the clinic: clinical-grade production, purification, and characterization of diverse oncolytic viruses.
Ungerechts G, Bossow S, Leuchs B, Holm PS, Rommelaere J, Coffey M, Coffin R, Bell J, Nettelbeck DM.
Mol Ther Methods Clin Dev. 2016 Apr 6;3:16018. doi: 10.1038/mtm.2016.18. eCollection 2016. Review.
Bioavailability, biodistribution, and CNS toxicity of clinical-grade parvovirus h1 after intravenous and intracerebral injection in rats.
Geletneky K, Leoni AL, Pohlmeyer-Esch G, Loebhard S, Leuchs B, Hoefer C, Jochims K, Dahm M, Huber B, Rommelaere J, Krebs O, Hajda J.
Comp Med. 2015 Feb;65(1):36-45.
Pathology, organ distribution, and immune response after single and repeated intravenous injection of rats with clinical-grade parvovirus h1.
Geletneky K, Leoni AL, Pohlmeyer-Esch G, Loebhard S, Baetz A, Leuchs B, Roscher M, Hoefer C, Jochims K, Dahm M, Huber B, Rommelaere J, Krebs O, Hajda J.
Comp Med. 2015 Feb;65(1):23-35.
Phase I/IIa study of intratumoral/intracerebral or intravenous/intracerebral administration of Parvovirus H-1 (ParvOryx) in patients with progressive primary or recurrent glioblastoma multiforme: ParvOryx01 protocol.
Geletneky K, Huesing J, Rommelaere J, Schlehofer JR, Leuchs B, Dahm M, Krebs O, von Knebel Doeberitz M, Huber B, Hajda J.
BMC Cancer. 2012 Mar 21;12:99. doi: 10.1186/1471-2407-12-99.
Full publication list
Angelova A, Barf M, Just A, Leuchs B, Rommelaere J, Ungerechts G.
Frehtman V, Wohlfarth D, Müller M, Krebs O, Leuchs B.
Wohlfarth D, Frehtman V, Müller M, Vogel M, Phan LMP, Brunecker A, Leuchs B.
Singhal M, Gengenbacher N, Abdul Pari AA, Kamiyama M, Hai L, Kuhn BJ, Kallenberg DM, Kulkarni SR, Camilli C, Preuß SF, Leuchs B, Mogler C, Espinet E, Besemfelder E, Heide D, Heikenwalder M, Sprick MR, Trumpp A, Krijgsveld J, Schlesner M, Hu J, Moss SE, Greenwood J, Augustin HG.
Hajda J, Leuchs B, Angelova AL, Frehtman V, Rommelaere J, Mertens M, Pilz M, Kieser M, Krebs O, Dahm M, Huber B, Engeland CE, Mavratzas A, Hohmann N, Schreiber J, Jäger D, Halama N, Sedlaczek O, Gaida MM, Daniel V, Springfeld C, Ungerechts G.
Neulinger-Muñoz M, Schaack D, Grekova SP, Bauer AS, Giese T, Salg GA, Espinet E, Leuchs B, Heller A, Nüesch JPF, Schenk M, Volkmar M, Giese NA.
Chen Q, Njenga R, Leuchs B, Chiocca S, Kleinschmidt J, Müller M.
Kiprianova I, Just A, Leuchs B, Rommelaere J, Angelova AL.
Bauer R, Enns H, Jungmann A, Leuchs B, Volz C, Schinkel S, Koch WJ, Raake PW, Most P, Katus HA, Müller OJ.
Lacroix J, Kis Z, Josupeit R, Schlund F, Stroh-Dege A, Frank-Stöhr M, Leuchs B, Schlehofer JR, Rommelaere J, Dinsart C.
Jungmann A, Leuchs B, Rommelaere J, Katus HA, Müller OJ.
Geiss C, Kis Z, Leuchs B, Frank-Stöhr M, Schlehofer JR, Rommelaere J, Dinsart C, Lacroix J.
Geletneky K, Hajda J, Angelova AL, Leuchs B, Capper D, Bartsch AJ, Neumann JO, Schöning T, Hüsing J, Beelte B, Kiprianova I, Roscher M, Bhat R, von Deimling A, Brück W, Just A, Frehtman V, Löbhard S, Terletskaia-Ladwig E, Fry J, Jochims K, Daniel V, Krebs O, Dahm M, Huber B, Unterberg A, Rommelaere J.
Hajda J, Lehmann M, Krebs O, Kieser M, Geletneky K, Jäger D, Dahm M, Huber B, Schöning T, Sedlaczek O, Stenzinger A, Halama N, Daniel V, Leuchs B, Angelova A, Rommelaere J, Engeland CE, Springfeld C, Ungerechts G.
Niopek K, Üstünel BE, Seitz S, Sakurai M, Zota A, Mattijssen F, Wang X, Sijmonsma T, Feuchter Y, Gail AM, Leuchs B, Niopek D, Staufer O, Brune M, Sticht C, Gretz N, Müller-Decker K, Hammes HP, Nawroth P, Fleming T, Conkright MD, Blüher M, Zeigerer A, Herzig S, Berriel Diaz M.
Leuchs B, Frehtman V, Riese M, Müller M, Rommelaere J.
Josupeit R, Bender S, Kern S, Leuchs B, Hielscher T, Herold-Mende C, Schlehofer JR, Dinsart C, Witt O, Rommelaere J, Lacroix J.
Ungerechts G, Bossow S, Leuchs B, Holm PS, Rommelaere J, Coffey M, Coffin R, Bell J, Nettelbeck DM.
Leuchs B, Roscher M, Müller M, Kürschner K, Rommelaere J.
Geletneky K, Leoni AL, Pohlmeyer-Esch G, Loebhard S, Leuchs B, Hoefer C, Jochims K, Dahm M, Huber B, Rommelaere J, Krebs O, Hajda J.
Geletneky K, Leoni AL, Pohlmeyer-Esch G, Loebhard S, Baetz A, Leuchs B, Roscher M, Hoefer C, Jochims K, Dahm M, Huber B, Rommelaere J, Krebs O, Hajda J.
Gardyan A, Osen W, Zörnig I, Podola L, Agarwal M, Aulmann S, Ruggiero E, Schmidt M, Halama N, Leuchs B, von Kalle C, Beckhove P, Schneeweiss A, Jäger D, Eichmüller SB.
Ayuso E, Blouin V, Lock M, McGorray S, Leon X, Alvira MR, Auricchio A, Bucher S, Chtarto A, Clark KR, Darmon C, Doria M, Fountain W, Gao G, Gao K, Giacca M, Kleinschmidt J, Leuchs B, Melas C, Mizukami H, Müller M, Noordman Y, Bockstael O, Ozawa K, Pythoud C, Sumaroka M, Surosky R, Tenenbaum L, van der Linden I, Weins B, Wright JF, Zhang X, Zentilin L, Bosch F, Snyder RO, Moullier P.
Alonso A, Reinz E, Leuchs B, Kleinschmidt J, Fatar M, Geers B, Lentacker I, Hennerici MG, de Smedt SC, Meairs S.
Werfel S, Jungmann A, Lehmann L, Ksienzyk J, Bekeredjian R, Kaya Z, Leuchs B, Nordheim A, Backs J, Engelhardt S, Katus HA, Müller OJ.
Lacroix J, Schlund F, Leuchs B, Adolph K, Sturm D, Bender S, Hielscher T, Pfister SM, Witt O, Rommelaere J, Schlehofer JR, Witt H.
Alonso A, Reinz E, Leuchs B, Kleinschmidt J, Fatar M, Geers B, Lentacker I, Hennerici MG, de Smedt SC, Meairs S.
Raykov Z, Grekova SP, Hörlein R, Leuchs B, Giese T, Giese NA, Rommelaere J, Zawatzky R, Daeffler L.
Jones A, Friedrich K, Rohm M, Schäfer M, Algire C, Kulozik P, Seibert O, Müller-Decker K, Sijmonsma T, Strzoda D, Sticht C, Gretz N, Dallinga-Thie GM, Leuchs B, Kögl M, Stremmel W, Diaz MB, Herzig S.
El-Andaloussi N, Leuchs B, Bonifati S, Rommelaere J, Marchini A.
Geletneky K, Huesing J, Rommelaere J, Schlehofer JR, Leuchs B, Dahm M, Krebs O, von Knebel Doeberitz M, Huber B, Hajda J.
Nieto K, Stahl-Hennig C, Leuchs B, Müller M, Gissmann L, Kleinschmidt JA.
Retargeting of rat parvovirus H-1PV to cancer cells through genetic engineering of the viral capsid.
Allaume X, El-Andaloussi N, Leuchs B, Bonifati S, Kulkarni A, Marttila T, Kaufmann JK, Nettelbeck DM, Kleinschmidt J, Rommelaere J, Marchini A.
Moehler M, Sieben M, Roth S, Springsguth F, Leuchs B, Zeidler M, Dinsart C, Rommelaere J, Galle PR.
Grekova SP, Aprahamian M, Daeffler L, Leuchs B, Angelova A, Giese T, Galabov A, Heller A, Giese NA, Rommelaere J, Raykov Z.
Pleger ST, Shan C, Ksienzyk J, Bekeredjian R, Boekstegers P, Hinkel R, Schinkel S, Leuchs B, Ludwig J, Qiu G, Weber C, Raake P, Koch WJ, Katus HA, Müller OJ, Most P.
Kiprianova I, Thomas N, Ayache A, Fischer M, Leuchs B, Klein M, Rommelaere J, Schlehofer JR.
Kaya Z, Leib C, Werfel S, Göser S, Öttl R, Leuchs B, Pfitzer G, Katus HA, Müller OJ.
Osen W, Soltek S, Song M, Leuchs B, Steitz J, Tüting T, Eichmüller SB, Nguyen XD, Schadendorf D, Paschen A.
El-Andaloussi N, Endele M, Leuchs B, Bonifati S, Kleinschmidt J, Rommelaere J, Marchini A.
Lock M, McGorray S, Auricchio A, Ayuso E, Beecham EJ, Blouin-Tavel V, Bosch F, Bose M, Byrne BJ, Caton T, Chiorini JA, Chtarto A, Clark KR, Conlon T, Darmon C, Doria M, Douar A, Flotte TR, Francis JD, Francois A, Giacca M, Korn MT, Korytov I, Leon X, Leuchs B, Lux G, Melas C, Mizukami H, Moullier P, Müller M, Ozawa K, Philipsberg T, Poulard K, Raupp C, Rivière C, Roosendaal SD, Samulski RJ, Soltys SM, Surosky R, Tenenbaum L, Thomas DL, van Montfort B, Veres G, Wright JF, Xu Y, Zelenaia O, Zentilin L, Snyder RO.
Ying Y, Müller OJ, Goehringer C, Leuchs B, Trepel M, Katus HA, Kleinschmidt JA.
Lacroix J, Leuchs B, Li J, Hristov G, Deubzer HE, Kulozik AE, Rommelaere J, Schlehofer JR, Witt O.
Nieto K, Kern A, Leuchs B, Gissmann L, Müller M, Kleinschmidt JA.
Angelova AL, Aprahamian M, Grekova SP, Hajri A, Leuchs B, Giese NA, Dinsart C, Herrmann A, Balboni G, Rommelaere J, Raykov Z.
Sieben M, Herzer K, Zeidler M, Heinrichs V, Leuchs B, Schuler M, Cornelis JJ, Galle PR, Rommelaere J, Moehler M.
Raykov Z, Grekova S, Leuchs B, Aprahamian M, Rommelaere J.
Sonntag F, Bleker S, Leuchs B, Fischer R, Kleinschmidt JA.
Abschuetz A, Kehl T, Geibig R, Leuchs B, Rommelaere J, Régnier-Vigouroux A.
Kuck D, Lau T, Leuchs B, Kern A, Müller M, Gissmann L, Kleinschmidt JA.
Müller OJ, Leuchs B, Pleger ST, Grimm D, Franz WM, Katus HA, Kleinschmidt JA.
Wilgenbus KK, Seranski P, Brown A, Leuchs B, Mincheva A, Lichter P, Poustka A.
Patents
Barbara Leuchs is inventor / joint inventor of granted patents
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