Journal: Cancers

Article Title: Adoptive Immunotherapy beyond CAR T-Cells

doi: 10.3390/cancers13040743

Figure Lengend Snippet: Evolution of the bioreactor platforms. The most advanced systems include G-Rex ® by Wilson Wolf, Z ® RP by Zellwerk, Xuri ® (WAVE ® ) by Cytiva, CliniMACS Prodigy ® by Miltenyi Biotec, Cocoon ® by Lonza (Basel, Switzerland), and Quantum ® by Terumo (Tokyo, Japan).

Article Snippet: The evolution of T-cell manufacturing systems began with the static technologies such as T-flasks and simple bag reactors, then proceeded to rocking motion (e.g., WAVE ® by Cytiva (Marlborough, MA, USA) bioreactors, and eventually evolved into PoC automated manufacturing systems such as CliniMACS Prodigy ® by Miltenyi Biotec (Bergisch Gladbach, Germany) ( ).

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Glutathione affinity chromatography for the scalable purification of an oncolytic virus immunotherapy from microcarrier cell culture

doi: 10.3389/fbioe.2023.1193454

Figure Lengend Snippet: Description of large-scale CVA21 cell culture in microcarrier single-use bioreactors and purification by glutathione affinity chromatography. Chromatography elution infectivity yield from clarified bulk and residual host cell DNA and bovine serum albumin concentrations shown. *Extrapolated HC-DNA concentration indicated if below the lower limit of quantitation, except when indistinguishable from no template controls.

Article Snippet: At the laboratory scale, MRC-5 cells were cultured in 3 L glass bioreactors with a BioFlo 320 controller (Cat#: 1379962911, Eppendorf, Hamburg, Germany) on 1 g/L Cytodex 1 microcarriers (Cat#: 17548702, Cytiva, Marlborough, MA) or in iCELLis Nano fixed-bed bioreactors (Pall Corporation, Port Washington, NY) with a 10 cm bed height with WMEM and 10% BCS controlled at a target temperature of 37°C and pH of 7.2 as described previously ( ).

Techniques: Cell Culture, Purification, Affinity Chromatography, Chromatography, Infection, Concentration Assay, Quantitation Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Glutathione affinity chromatography for the scalable purification of an oncolytic virus immunotherapy from microcarrier cell culture

doi: 10.3389/fbioe.2023.1193454

Figure Lengend Snippet: Optimization of microcarrier bioreactor temperature during CVA21 infection. (A) Relative infectivity sampled across 4–5 days post-infection controlled at 34°C (blue diamond), 35.5°C (green square), 37°C (yellow triangle), and 38.5°C (red circle). Error bars represent one standard deviation of an average of two replicate conditions. (B) Average peak relative infectivity, defined as the highest sample infectivity across the 4–5-day bioreactor infection phase, as a function of infection temperature across three experiments. Error bars represent one standard deviation of an average at each temperature condition. See for a description of experimental replicates.

Article Snippet: At the laboratory scale, MRC-5 cells were cultured in 3 L glass bioreactors with a BioFlo 320 controller (Cat#: 1379962911, Eppendorf, Hamburg, Germany) on 1 g/L Cytodex 1 microcarriers (Cat#: 17548702, Cytiva, Marlborough, MA) or in iCELLis Nano fixed-bed bioreactors (Pall Corporation, Port Washington, NY) with a 10 cm bed height with WMEM and 10% BCS controlled at a target temperature of 37°C and pH of 7.2 as described previously ( ).

Techniques: Infection, Standard Deviation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Glutathione affinity chromatography for the scalable purification of an oncolytic virus immunotherapy from microcarrier cell culture

doi: 10.3389/fbioe.2023.1193454

Figure Lengend Snippet: Impact of cell culture controlled at 37°C and 34°C during infection on downstream GSH affinity chromatography performance. (A) SDS-PAGE of CB (100x pre-dilution) and GSH elution (no pre-dilution) harvested from bioreactors controlled at 37°C (lane 1–2) and 34°C (lane 3–4) from experiment 1. (B) GSH elution sample relative ratio of the 34°C–37°C material from experiment 1 (green, horizontal bar) and experiment 2 (blue, dotted) for infectious particles by relative infectivity assay and total genomes by RT-qPCR. (C) GSH elution sample % empty capsid content by RP-UPLC for the 34°C and 37°C materials from experiment 1 (green, horizontal bar) and experiment 2 (blue, dotted).

Article Snippet: At the laboratory scale, MRC-5 cells were cultured in 3 L glass bioreactors with a BioFlo 320 controller (Cat#: 1379962911, Eppendorf, Hamburg, Germany) on 1 g/L Cytodex 1 microcarriers (Cat#: 17548702, Cytiva, Marlborough, MA) or in iCELLis Nano fixed-bed bioreactors (Pall Corporation, Port Washington, NY) with a 10 cm bed height with WMEM and 10% BCS controlled at a target temperature of 37°C and pH of 7.2 as described previously ( ).

Techniques: Cell Culture, Infection, Affinity Chromatography, SDS Page, Quantitative RT-PCR

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Glutathione affinity chromatography for the scalable purification of an oncolytic virus immunotherapy from microcarrier cell culture

doi: 10.3389/fbioe.2023.1193454

Figure Lengend Snippet: Characterization of GSH chromatography elution purified from large-scale microcarrier bioreactors controlled at 37°C (yellow, horizontal bar) and 34°C (blue, dotted) during infection. Data represent average values from batches shown in , and error bars represent one standard deviation. (A) Relative comparison of batch productivity at GSH elution. (B) Comparison of viral genomes by RT-qPCR and viral particles by VP1 CE Western assays at GSH elution. (C) Comparison of % empty capsids by RP-UPLC in the GSH elution.

Article Snippet: At the laboratory scale, MRC-5 cells were cultured in 3 L glass bioreactors with a BioFlo 320 controller (Cat#: 1379962911, Eppendorf, Hamburg, Germany) on 1 g/L Cytodex 1 microcarriers (Cat#: 17548702, Cytiva, Marlborough, MA) or in iCELLis Nano fixed-bed bioreactors (Pall Corporation, Port Washington, NY) with a 10 cm bed height with WMEM and 10% BCS controlled at a target temperature of 37°C and pH of 7.2 as described previously ( ).

Techniques: Chromatography, Purification, Infection, Standard Deviation, Comparison, Quantitative RT-PCR, Western Blot

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: Description of iCELLis Nano bioreactor runs for continuous LV production process development

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques:

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: Growth kinetics and cell distribution WinPac-RDpro-GFP (A) cell density and (B) cell viability in the iCELLis Nano bioreactor runs. Three macro-carriers were samples from each bioreactor. (C) Distribution of WinPac-RDpro-GFP cells in the iCELLis Nano bioreactor fixed-bed at day 10 of runs 6–8 as measured from the top, middle, and bottom macro-carriers. Points represent mean value ± 1 SD (n = 3).

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques:

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: Specific growth rate, doubling time and population doublings of WinPac-RDpro-GFP cells during the iCELLis Nano bioreactor runs

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques:

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: Metabolite concentrations Daily offline measurement of (A) glucose, (B) lactate, and (D) ammonium during the iCELLis Nano bioreactor runs. Points represent mean value ± 1 SD (n = 1). (C) The lactate yield from glucose for each iCELLis Nano bioreactor runs from days one to ten. Bars represent mean value ± 1 SD (n = 3). The 2 mol mol −1 reference line indicates the maximum theoretical yield of lactate from glucose from anaerobic glycolysis.

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques:

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: Infectious and physical LV yields obtained from the iCELLis Nano bioreactor runs

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques:

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: Cell specific lentiviral vector titres Daily specific productivity of (A) infectious and (B) physical LVs during the iCELLis Nano bioreactor runs. Bars represent mean value ± 1 SD.

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques: Plasmid Preparation

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Continuous manufacturing of lentiviral vectors using a stable producer cell line in a fixed-bed bioreactor

doi: 10.1016/j.omtm.2024.101209

Figure Lengend Snippet: dsDNA concentration Pre-clarification (A) intracellular dsDNA released on a per cell basis and (B) total DNA concentration across the iCELLis Nano bioreactor processes. Bars represent mean value ± 1 SD.

Article Snippet: The iCELLis Nano bioreactor, equipped with a 2 cm low-compaction fixed-bed (0.53 m 2 surface area) and the mPath bioreactor benchtop control tower (Cytiva), was used.

Techniques: Concentration Assay, Clarification Assay