Endotoxin and Microbiological Control

Williams, Kevin L.

doi:10.1007/978-3-030-17148-3_5

Cited by 2 publications

(2 citation statements)

References 78 publications

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“…Current protocols attempt to increase the maturation of DCs by multiple compounds, which often target the pattern recognition receptors, namely TLRs [ 44 ]. However, many of these, often highly potent, compounds, such as lipopolysaccharides (LPS), are not permitted for the maturation of DCs produced for clinical use, mainly due to endotoxin level regulation [ 45 ]. Their replacement for other compounds may sometimes provide a comparable maturation-inducing efficacy [ 46 ].…”

Section: Discussionmentioning

confidence: 99%

Thapsigargin-Stimulated LAD2 Human Mast Cell Line Is a Potent Cellular Adjuvant for the Maturation of Monocyte-Derived Dendritic Cells for Adoptive Cellular Immunotherapy

Taborska

Stakheev

Bartůňková

et al. 2021

IJMS

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The preparation of dendritic cells (DCs) for adoptive cellular immunotherapy (ACI) requires the maturation of ex vivo-produced immature(i) DCs. This maturation ensures that the antigen presentation triggers an immune response towards the antigen-expressing cells. Although there is a large number of maturation agents capable of inducing strong DC maturation, there is still only a very limited number of these agents approved for use in the production of DCs for ACI. In seeking novel DC maturation agents, we used differentially activated human mast cell (MC) line LAD2 as a cellular adjuvant to elicit or modulate the maturation of ex vivo-produced monocyte-derived iDCs. We found that co-culture of iDCs with differentially activated LAD2 MCs in serum-containing media significantly modulated polyinosinic:polycytidylic acid (poly I:C)-elicited DC maturation as determined through the surface expression of the maturation markers CD80, CD83, CD86, and human leukocyte antigen(HLA)-DR. Once iDCs were generated in serum-free conditions, they became refractory to the maturation with poly I:C, and the LAD2 MC modulatory potential was minimized. However, the maturation-refractory phenotype of the serum-free generated iDCs was largely overcome by co-culture with thapsigargin-stimulated LAD2 MCs. Our data suggest that differentially stimulated mast cells could be novel and highly potent cellular adjuvants for the maturation of DCs for ACI.

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Section: Discussionmentioning

confidence: 99%

Thapsigargin-Stimulated LAD2 Human Mast Cell Line Is a Potent Cellular Adjuvant for the Maturation of Monocyte-Derived Dendritic Cells for Adoptive Cellular Immunotherapy

Taborska

Stakheev

Bartůňková

et al. 2021

IJMS

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“…1 Endotoxins are typically quantified by the Limulus amebocyte lysate (LAL) assay as recommended by the FDA and Pharmacopeias. 20 However, low endotoxin recovery (LER) is an issue with this assay, which can result in a false negative. 21,22 Alkaline lysis treatment of fermented plasmid harvest releases endotoxins into the lysate.…”

Section: The Challenge Of Endotoxinsmentioning

confidence: 99%

Improved endotoxin removal using ecofriendly detergents for intensified plasmid capture

Kiesewetter,

Gupta,

Heinen‐Kreuzig

et al. 2023

Biotechnology Progress

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Increasing plasmid demand for both production of viral and gene therapies as well as nucleic acid based vaccines has highlighted bottlenecks in production. One bottleneck is traditional bead‐based chromatography as a capture step. To meet the needs of fast‐growing markets, new production solutions are needed. These solutions must enable efficient capture of a diverse range of plasmid types and excellent clearance of bacterial host impurities, such as endotoxin. Enhanced endotoxin clearance during chromatographic purification has previously been demonstrated with detergents such as Triton™ X‐100. However, degradation products of Triton™ X‐100 are known to have a negative environmental impact, and more sustainable, environmentally benign alternatives have been identified. This work establishes an efficient, intensified plasmid capture using convective anion exchange (AEX) chromatography. The feasibility of the intensified capture approach was assessed with different membrane and a monolith AEX supports. Various detergents from different physico‐chemical classes were evaluated with different AEX technologies. Purification efficiency evaluated endotoxin and host cell protein (HCP) clearance, plasmid yield, potential interference of the detergents with analytical in‐process control assays, and overall process compatibility. This comprehensive screening approach provides valuable insights to intensified plasmid production.

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Endotoxin and Microbiological Control

Cited by 2 publications

References 78 publications

Thapsigargin-Stimulated LAD2 Human Mast Cell Line Is a Potent Cellular Adjuvant for the Maturation of Monocyte-Derived Dendritic Cells for Adoptive Cellular Immunotherapy

Thapsigargin-Stimulated LAD2 Human Mast Cell Line Is a Potent Cellular Adjuvant for the Maturation of Monocyte-Derived Dendritic Cells for Adoptive Cellular Immunotherapy

Improved endotoxin removal using ecofriendly detergents for intensified plasmid capture

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