Shenzhen Cell Valley achieved a major breakthrough in CAR-NK preparation process
In view of the pain points in the early development and industrial production of CAR-NK cells, the core R&D team of Shenzhen Cell Valley conducted long-term research on its underlying technology, and finally made a breakthrough in the preparation process of CAR-NK cells: NK cells were transduced by retrovirus vector produced by PackRV-SS system unique to Shenzhen Cell Valley. The transduction efficiency was stable above 60%, and the positive rate remained stable. Moreover, CAR-NK cells proliferated up to 10,000 times within 30 days of culture after transduction. This breakthrough will greatly promote the process of product development and clinical application of CAR-NK cells. Enterprises and institutions engaged in NK and CAR-NK cell therapy are welcome to cooperate with Shenzhen Cell Valley to jointly promote the development of the industry.
In the past two years, in addition to CAR-T therapy has attracted much attention, another new immune cell therapy - natural killer (NK) cell therapy is also regarded as a new generation of anti-tumor special treatment of solid tumors. However, the main problems facing the current CAR-NK production are: the efficiency of exogenous gene transduction is low and unstable; NK cell culture is difficult, in vitro proliferation multiple is low, can not meet the clinical treatment of cell dosage. The amount of initial virus vector is large and the cost is high. The retrovirus with novel envelope design can achieve efficient transduction of primary NK cells, and achieve stable, economical and large-scale retrovirus vector production through a stable virus-producing monoclonal packaging cell line.
The Cell Valley Core research and development team in Shenzhen used a retrovirus vector designed with a new envelope to transduce NK cells, and detected a CAR-NK positive rate of 67.7% after 2 days of viral transduction. In addition, the positive rate of CAR-NK was continuously detected until the 27th day after virus transduction, and the positive rate of CAR-NK remained at 71%, at a stable and high transduction level. At the same time, after 10 days of transduction, the purity of NK cells (CD56+CD3-) in CAR-NK cells remained above 95%, and the content of T cells (CD56-CD3+) was less than 0.5%. The purity is very high, has reached and far exceeded the United States FDA recognized allograft standards (NK cells greater than 90%, T cells less than 5%), can avoid the occurrence of GVHD.
三、The killing effect of CAR-NK cells in vitro was better than that of NK cells at low target ratio (1:1 and 1:3)
In the past two years, in addition to CAR-T therapy has attracted much attention, another new immune cell therapy - natural killer (NK) cell therapy is also regarded as a new generation of anti-tumor special treatment of solid tumors. However, the main problems facing the current CAR-NK production are: the efficiency of exogenous gene transduction is low and unstable; NK cell culture is difficult, in vitro proliferation multiple is low, can not meet the clinical treatment of cell dosage. The amount of initial virus vector is large and the cost is high. The retrovirus with novel envelope design can achieve efficient transduction of primary NK cells, and achieve stable, economical and large-scale retrovirus vector production through a stable virus-producing monoclonal packaging cell line.
一、The single transduction positive rate of CAR-NK reached more than 60% and remained at a relatively stable level for a long time, and there was no situation of CAR positive rate decreasing
The Cell Valley Core research and development team in Shenzhen used a retrovirus vector designed with a new envelope to transduce NK cells, and detected a CAR-NK positive rate of 67.7% after 2 days of viral transduction. In addition, the positive rate of CAR-NK was continuously detected until the 27th day after virus transduction, and the positive rate of CAR-NK remained at 71%, at a stable and high transduction level. At the same time, after 10 days of transduction, the purity of NK cells (CD56+CD3-) in CAR-NK cells remained above 95%, and the content of T cells (CD56-CD3+) was less than 0.5%. The purity is very high, has reached and far exceeded the United States FDA recognized allograft standards (NK cells greater than 90%, T cells less than 5%), can avoid the occurrence of GVHD.
二、The proliferation of CAR-NK is more than 10,000 times, which can meet the needs of cell consumption in non-clinical trials and even clinical trials
The research team continuously monitored the proliferation status of the untransduced NK cells as well as the CAR NK cells after transduction. The proliferation of NK cells reached 15,000-fold after 32 days of culture, and 10,000-fold after 27 days of CAR-NK cell transduction. Both the proliferation of NK and CAR-NK can meet the cell dosage of commercially available NK cell therapy. In the continuous expansion, the purity of NK cells was always greater than 96%. The positive rate of CAR in CAR-NK was always greater than 65% in one month, overcoming the problem of declining CAR positive rate reported by other NK cell research institutions.
三、The killing effect of CAR-NK cells in vitro was better than that of NK cells at low target ratio (1:1 and 1:3)
The research team also conducted a preliminary assessment of the killing ability of NK and CAR-NK cells in vitro. The experimental results showed that there was no significant difference in the killing effect of NK and CAR-NK at the high efficiency target ratio (3:1 and 10:1), and the killing efficiency of both on target cells reached more than 90%. At the low efficiency target ratio (1:3 and 1:1), the killing effect of CAR-NK was better than that of NK cells. Compared with CAR-T, the killing effect of CAR-NK was superior to that of CAR-T cells in most target ratios (1:1, 3:1 and 10:1). These data indicate that NK transduction by retroviral vector can enhance the killing ability of NK cells in vitro to a certain extent.
Shenzhen Cell Valley has designed and produced CAR-specific retroviral vector products for a variety of tumor targets such as BCMA, CD19, CD38, CD22, Trop2, FRα, CD7, CD33, EGFR/EGFRVIII, etc., and all of them have achieved efficient transduction of human primary T cells. Further research will also be carried out in the field of immune cell therapy such as NK cells, γδT cells and macrophages. Shenzhen Cell Valley warmly welcomes enterprises and institutions in the CGT field to exchange and guide their work, and make use of the huge advantages of retrovirus vectors in scientific research and industrial production to jointly develop cell products based on retrovirus vectors and jointly promote the advent of new CAR-NK cell products.
【 At present, Shenzhen Cell Valley can provide trial packages of CD19-CAR and GFP-Luciferase virus products prepared based on retroviral vector for testing. You can contact the Marketing Department and Sales Department at any time to obtain trial packages. 】
Disclaimer: Shenzhen Cell Valley is committed to the research of cell and gene therapy, in order to promote emerging technologies, so that more people understand the new development of biomedicine. The content of this article is only used for information exchange, and the platform remains neutral on the content, statements and opinions of the article, and does not represent the position and views of Shenzhen Cell Valley. The relevant information in this article should not be used as a diagnosis or treatment, is not a substitute for professional medical advice, and the company's website will not assume any responsibility. The final interpretation of the content of the above statement belongs to the company's website, this statement will apply to the company's website all the time to share the article, thank you for your cooperation! Copyright description: The copyright of the article belongs to Shenzhen Cell Valley, individuals are welcome to forward to the circle of friends, media or institutions without authorization, reproduced in any form to other platforms, will be regarded as infringement. For reprinting, please contact email: contact@sz-cell.com
Shenzhen Cell Valley has designed and produced CAR-specific retroviral vector products for a variety of tumor targets such as BCMA, CD19, CD38, CD22, Trop2, FRα, CD7, CD33, EGFR/EGFRVIII, etc., and all of them have achieved efficient transduction of human primary T cells. Further research will also be carried out in the field of immune cell therapy such as NK cells, γδT cells and macrophages. Shenzhen Cell Valley warmly welcomes enterprises and institutions in the CGT field to exchange and guide their work, and make use of the huge advantages of retrovirus vectors in scientific research and industrial production to jointly develop cell products based on retrovirus vectors and jointly promote the advent of new CAR-NK cell products.
【 At present, Shenzhen Cell Valley can provide trial packages of CD19-CAR and GFP-Luciferase virus products prepared based on retroviral vector for testing. You can contact the Marketing Department and Sales Department at any time to obtain trial packages. 】
Disclaimer: Shenzhen Cell Valley is committed to the research of cell and gene therapy, in order to promote emerging technologies, so that more people understand the new development of biomedicine. The content of this article is only used for information exchange, and the platform remains neutral on the content, statements and opinions of the article, and does not represent the position and views of Shenzhen Cell Valley. The relevant information in this article should not be used as a diagnosis or treatment, is not a substitute for professional medical advice, and the company's website will not assume any responsibility. The final interpretation of the content of the above statement belongs to the company's website, this statement will apply to the company's website all the time to share the article, thank you for your cooperation! Copyright description: The copyright of the article belongs to Shenzhen Cell Valley, individuals are welcome to forward to the circle of friends, media or institutions without authorization, reproduced in any form to other platforms, will be regarded as infringement. For reprinting, please contact email: contact@sz-cell.com
