China
Africa
Explore chapters and articles related to this topic
Production of Clinical Lots of Gene Therapy Vectors Using Good Manufacturing Practice: Experience in a University Setting
Published in Eric Wickstrom, Clinical Trials of Genetic Therapy with Antisense DNA and DNA Vectors, 2020
The quality assurance tests for the Production Lots may include some of the same tests and may also include the Limulns amebocyte lysate test. The purpose of this test is to detect and quantitatively determine the gram-negative bacterial endotoxin level of a test article or extract using the Limulus Amebocyte Lysate (LAL) gel-clot method for testing. Quality assurance tests can also be performed on the recombinant adenovirus lysates. Establishment of a Master Cell Bank requires extensive testing. Quality assurance tests performed on the Master Cell Bank might include mycoplasma and sterility testing as well as cell culture identification.
Biologic Drug Substance and Drug Product Manufacture
Published in Anthony J. Hickey, Sandro R.P. da Rocha, Pharmaceutical Inhalation Aerosol Technology, 2019
Ajit S. Narang, Mary E. Krause, Shelly Pizarro, Joon Chong Yee
CHO cells have been grown at high cell densities up to >30 × 106 cells/mL in large-scale production vessels (>15 kilo-liters). The upstream manufacturing process initiates with a vial thaw of a cell bank (Figure 8.2), and cells are expanded in progressively higher volumes in culture vessels such as shake flasks and/or wave bioreactors. Prior to inoculation of large production vessels, the cells are expanded through a series of inoculum train bioreactors. Depending on the scale of the production bioreactor, this upstream process may take 2 weeks–6 weeks from vial thaw.
CMC Requirements for Biological Products
Published in Shein-Chung Chow, Analytical Similarity Assessment in Biosimilar Product Development, 2018
The CMC development for biological drug products starts with establishment of the expression system. A cell-line will be selected among bacterial, yeast, and mammalian host strains, and then the correct DNA sequence will be inserted. Elaborate cell-screening and selection methods are then used to establish a master cell bank. Extensive characterization on the master cell bank needs to be carried out to provide microbiological purity or sterility and identity (CBER/FDA, 1993). Bulk protein production involves developing robust and scalable fermentation and purification processes.
miR-761-hepcidin/Gpx4 pathway contribute to unexplained liver dysfunction in polycystic ovary syndrome by regulating liver iron overload and ferroptosis
Published in Gynecological Endocrinology, 2023
Ruoheng Zheng, Chuanping Lin, Yuchan Mao, Fan Jin
Cells were purchased from cell bank of the Chinese academy of sciences (shanghai). AML12 cells were cultured in 10 μg/mL insulin, 10 μg/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone, 100 U/mL penicillin and 100 mg/mL streptomycin, 10% FBS in DMEM/F-12 medium at 37 °C in a 5% CO2 incubator. L02 cells were routinely cultured in DMEM medium with 100 mg/mL penicillin and 100 mg/mL streptomycin in the same condition. A model of hepatocytes interfering/overexpressing miR-761 was established in AML2 cells by adding miR-761 mimics (50 nM) or inhibitors (100 nM) and were collected 24 h after transfection. AML12 cells were divided into 4 groups: Control group, negative control (NC) group, miR-761-inhibitor group and miR-761-mimic group. L02 cells were incubated with miR-761 mimics for 24 h, followed by ferroptosis inhibitor-Ferrostatin-1 (2 μM) and Fe chelator-DFO (1 mM) addition for 24 h and 48 h. They were divided into 6 groups: Ctrl group (A), NC mic group (B), miR-761-mic group (C), DMSO + miR-761-mic group (D), DFO + miR-761-mic group (E) and Fer-1 + miR-761-mic group (F). The mRNA levels of miR-761, TNF-α; IL-6, IL-1β by qRT-PCR; the protein levels of TNF-α; IL-6; IL-1β by ELISA and GPX4/hepcidin levels by western blot were routinely detected.
Generation and functional characterization of a multigene-modified NK101 cell line exerting diverse mechanisms of antitumor action
Published in OncoImmunology, 2022
Injung Hwang, Hyun Tak Jin, Moon Cheol Kang, Tae Yoon Kim, Young Chul Sung, Sae Won Kim
The integration of genetic engineering technologies into cellular immunotherapy has facilitated rapid advancement of the immuno-oncology field, as represented by landmark approvals of patient-derived T cells modified to express a chimeric receptor that redirects cellular specificity to a tumor antigen.1 Driven by the success of the first-generation therapeutics, growing efforts are being directed toward implementing multiple genetic modifications to alter or enhance various functional traits of antitumor immune cells and identifying an optimal cell source/type to apply such engineering.1,2 As the insertion or deletion of multiple genes accompanies increased manufacturing and regulatory complexities, potentially necessitating longer ex vivo processing and more comprehensive characterization of the final products, primary immune cells that are prone to terminal differentiation/exhaustion with limited time frame for efficient genetic manipulation would be challenging targets for multigenic modification.3–5 In this regard, cell line-based platforms, such as clonal master induced pluripotent stem cell (iPSC) lines and immortalized natural killer (NK) cell lines, serve as an attractive alternative source for multi-functional or multi-step engineering, as (i) they can be easily grown and repeatedly manipulated in vitro; (ii) single cells stably expressing multiple transgenes can be clonally selected for the creation of a master cell bank; and (iii) standardized and fully characterized products consisting of defined populations of therapeutic cells can be manufactured at clinical scale.6
Effects of the COVID-19 pandemic: new approaches for accelerated delivery of gene to first-in-human CMC data for recombinant proteins
Published in mAbs, 2023
Hervé Broly, Jonathan Souquet, Alain Beck
In conclusion, as of today, the use of a non-clonally-derived master cell bank to produce materials for human studies could be acceptable solely in a specific emergency context, or in case of a product fulfilling an unmet high medical need and where a program exhibits a high benefit-to-risk ratio provided that: 1) the RCB has been established in compliance with GMP and tested for adventitious agents in accordance with appropriate ICH guidelines, and 2) the residual uncertainty of the impact of non-clonality is counterbalanced by testing for genetic stability of aged cells and implementing an augmented control strategy, such as batch-to-batch amino acid sequence variant analysis.