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Your Position: Home > CMC Production Process of Adoptive NK Cell Therapies
Adoptive cell therapy (ACT) is a promising strategy in cancer immunotherapy, aiming to enhance immune function through ex vivo modification and expansion of immune cells before reinfusion into patients. Apart from chimeric antigen receptor T-cell (CAR-T) therapy, newer forms of ACT such as T-cell receptor T-cell (TCR-T) therapy, tumor-infiltrating lymphocyte (TIL) therapy, chimeric antigen receptor natural killer (CAR-NK) cell therapy, and chimeric antigen receptor macrophage (CAR-M) therapy have emerged in clinical settings.
CAR-NK cell therapy, as a novel treatment approach, offers several advantages over CAR-T therapy: 1) Faster efficacy: without the need for antigen presentation, 2) Higher safety profile: reduced risk of graft-versus-host disease (GVHD) and cytokine release syndrome (CRS), and 3) Effectiveness against both hematologic malignancies and solid tumors. However, CAR-NK faces a key challenge: obtaining enough NK cells for cell manufacturing.
To support research and CMC manufacturing processes related to cell therapies, we have developed a series of CMC production process related products for CAR-NK, helping customers accelerate the production process and fully supporting the development process based on adoptive NK Cell Therapies.

CMC Production Process of Adoptive NK Cell Therapies

The manufacturing process of CAR-NK cells involves several steps: NK cell selection, cell isolation, cell activation, CAR construction, cell expansion, cryopreservation, and infusion back into patients. Each step in this process requires strict adherence to Good Manufacturing Practice (GMP) standards.
CMC Production Process of Adoptive NK Cell Therapies
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               Activation and expansion of NK cells from different sources

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               Enhancement of NK cell activity

Activation and expansion of NK cells from different sources

Cytokines play a crucial role in the maturation, activation, survival, and functionality of NK cells, making them a fundamental component of the ex vivo NK cell maintenance system. NK cells can be derived from a wide range of sources. Various types of NK cells, such as induced pluripotent stem cells (iPSCs), embryonic stem cells (ESCs), peripheral blood mononuclear cells (PBMCs), umbilical cord blood (UCB) cells, and NK cell lines, all of which are used in ex vivo NK cell production.

NK cell activation products

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  • iPSC/ESC Source

  • PBMC Source

  • UCB Source

  • NK Cell Lines Source

iPSC/ESC Source

iPSC-NK differentiation culture can be conducted using feeder or feeder-free systems. Feeder-free cultures, particularly those using recombinant DLL4 protein, are becoming the mainstream for clinical-scale iPSC production due to concerns over batch-to-batch consistency and safety with feeder cells derived from animal sources.

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Product Features

GMP Grade DLL4, VCAM1:Eliminate the need of feeder cells in culture systems and support differentiation of iPSCs to NK cells

Validated activity by iPSC/ESC to NK cell differentiation

Production and quality control are carried out under strict GMP systems and comply with regulations from multiple countries

Comprehensive quality release verification, with 16 quality control indicators

Enhanced safety (sterile, no mycoplasma, no exogenous virus, animal free production system, and various impurities residual detection)

Produced in a Pharmaceutical-grade production facility

Support for online and offline audits

Completion of FDA Drug Master File (DMF) registration

Validation Data

DLL4 & VCAM-1 for NK differentiation
DLL4 & VCAM-1 for NK differentiation

The combination of DLL4 (GMP-DL4H28) & VCAM1 (GMP-VC1H25) could significantly facilitate the differentiation efficiency of CD56+ CD3- NK cells.

SCF, FLT3L, IL7 for NK differentiation
DLL4 & VCAM-1 for NK differentiation

SCF(GMP-SCFH25), Flt3L(GMP-FLLH28), IL-7(GMP-L07H24) could significantly promote the HSPC differentiation to NK cells, comparable to Company P.

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PBMCs are advantageous for NK cell therapy due to their ease of collection, ability to expand in vitro, and low toxicity. However, NK cells constitute only 10-15% of PBMCs, necessitating the use of cytokines to stimulate NK cell proliferation. Studies suggest that combinations of multiple cytokines are more effective than single cytokines in inducing NK cell expansion.

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Recommended cytokines combination:IL-2+IL-15  IL-15+IL-21  IL-2+IL-18

Product Features

Validated activity by PBMC to NK cell differentiation

Production and quality control are carried out under strict GMP systems and comply with regulations from multiple countries

Comprehensive quality release verification, with 16 quality control indicators

Enhanced safety (sterile, no mycoplasma, no exogenous virus, animal free production system, and various impurities residual detection)

Produced in a Pharmaceutical-grade production facility

Support for online and offline audits

Completion of FDA Drug Master File (DMF) registration

Validation Data

Human PBMCs were cultured with IL-2

Human PBMCs were cultured with IL-2 (Cat. No. GMP-L02H14), IL-15 (Cat. No. GMP-L15H13), IL-18 (Cat. No. IL8-H5114), IL-21 (Cat. No. GMP-L21H25). The result shows that they are functional and can promote the expansion of these cells with a reasonable cell viability.

Human PBMCs were cultured with IL-2

Human PBMCs were cultured with IL-2 (Cat. No. GMP-L02H14), IL-15 (Cat. No. GMP-L15H13), IL-18 (Cat. No. IL8-H5114), IL-21 (Cat. No. GMP-L21H25). The result shows that they are functional and can increase the percentage of the CD3-CD56+ cells.

iPSC-NK

Large quantities of NK cells can be obtained from UCB through two main approaches: direct expansion of NK cells from UCB or differentiation of CD34+ hematopoietic stem and progenitor cells from UCB into NK cells followed by expansion.

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Product Features

Validated activity by UCB to NK cell differentiation

Production and quality control are carried out under strict GMP systems and comply with regulations from multiple countries

Comprehensive quality release verification, with 16 quality control indicators

Enhanced safety (sterile, no mycoplasma, no exogenous virus, animal free production system, and various impurities residual detection)

Pharmaceutical-grade production facility

Support for online and offline audits

Completion of FDA Drug Master File (DMF) registration

Currently, besides NK-92 cell line used in clinical research, other NK cell lines are in preclinical stages. NK cell lines are typically expanded using cytokine-dependent culture systems, where all NK cell lines can proliferate in IL-2-containing media.

Product List

Product Features

Production and quality control are carried out under strict GMP systems and comply with regulations from multiple countries

Comprehensive quality release verification, with 16 quality control indicators

Enhanced safety (sterile, no mycoplasma, no exogenous virus, animal free production system, and various impurities residual detection)

Pharmaceutical-grade production facility

Support for online and offline audits

Completion of FDA Drug Master File (DMF) registration

Enhancement of NK cell activity

Clinical trials have shown that allogeneic NK cells typically survive for only a few weeks after injection. Adoptive NK cells have limited proliferation in vivo, leading to poor clinical outcomes against solid tumors. Modifying genes related to NK cell metabolism and replication using CRISPR-Cas9 technology can promote the persistence of NK cells in vivo and enhance their anti-tumor activity. Additionally, pre-activating NK cells with a cocktail of cytokines (IL-12, IL-15, and IL-18) can also improve their persistence and impart a memory-like phenotype to NK cells.

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Product Features

High purity, high enzyme activity, high cleavage efficiency

Possesses nuclear localization signals to enhance editing efficiency

Aseptic, ultra-low endotoxin

Produced in GMP-compliant facilities and undergoes QC testing

Validation Data

GMP FGF basic-Stemness maintenance

Different amounts of Cas9 were incubated with the same amount of excess gRNA and plasmid for 60 minutes at 37°C. When using 400-200 ng Acro Cas9, the cutting efficiency is greater than 90%. In comparison, when using a 200 ng Competitor T, the cutting efficiency is only about 50%.

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