CAR-T Lentiviral Particles Helping to Establish an Efficient and Simple Platform for CAR-T Cell Expansion and Generation

Chimeric antigen receptor (CAR) T-cells represent a paradigm shift in cancer immunotherapy and a new milestone in the history of oncology. Among them, CD19-targeted drugs have achieved unprecedented success in the treatment of B-cell malignancies and were approved by the US FDA in 2017. The CAR approach is currently being evaluated in multiple pivotal trials designed for the immunotherapy of hematological malignancies as well as solid tumors.

CAR-T Process Overview

The production of autologous CAR T cells is carried out by a variety of manufacturing approaches all comprising the same common steps. First, the patient's white blood cells (WBCs) are isolated by leukapheresis and washed. Then, the T cells are activated, transduced with the CAR transgene, expanded to the required cell numbers for therapy, formulated and filled. After quality control testing and preparatory lymphodepleting chemotherapy for the patient, the product is injected into the patient.

Fig. 1 The production of autologous CAR T cells. (Vormittag, et al., 2018)

How to Generate CAR-T Cells using Lentiviral Vectors?

Lentiviral Vectors (LVs) are produced by transfection of HEK293T cells and purified from the cell culture supernatant. Autologous T-cells are isolated and activated prior to transduction. The CAR transgene is delivered into the activated T-cells by LVs, and then expanded. Finally, produced CAR T-cells are formulated in an adjusted buffer in a defined ratio of CD4þ:CD8þ CAR T-cell.

Fig. 2 Flow scheme of the CAR T cell manufacturing process using LVs. (Poorebrahim, et al.; 2019)

How to Achieve Efficient Transduction of CAR?

At present, CAR-T cell therapies are facing two major problems: difficult to scale up and high cost. Therefore, it is of great practical importance to reduce the production cost, achieve large-scale production, and shorten the production cycle for the popularization of CAR-T. The production and quality control of gene delivery vectors for CAR-T, a central aspect of CAR-T technology, also have an extremely important impact on CAR-T therapies.

In order to effectively introduce CAR genes into T lymphocytes and construct high-quality CAR-T cells, efficient gene introduction efficiency is of course crucial. It is well known that lentiviral vectors are the main gene introduction platform used in the current research of transmissible T cell therapy. Lentiviral vectors are generally developed based on HIV-1 (Human immunodeficiency virus type 1) and can be used for gene introduction into most mammalian cells, including non-dividing cells such as hematopoietic stem cells and neuronal cells, which are difficult to achieve using retroviral vectors.

To generate CAR-T cells in vitro, Creative Biogene offers a wide range of stock CAR-T lentiviral particles targeting human CD19, CD20, CD33, CD38, CEA, EGFR, EPCAM, GPC3, HER2, MSLN, PSMA, TACSTD2, etc., and this range is constantly being enriched and updated. These lentiviral vectors (LVs) are particularly attractive because of their ability to stably integrate relatively large DNA inserts and efficiently transduce dividing and non-dividing cells.

Benefits of Selecting Lentiviral Vectors for CAR-T Cell Transduction

The viral vector used to transduce CARs into T cells is considered a key raw material in the CAR-T cell production process. Lentiviruses are currently used as vectors for infecting T cells in the vast majority of the cellular immunotherapy pipeline under development worldwide. All of the marketed CAR-T drugs targeting CD19 and BCMA, without exception, use lentiviruses as their vectors. Gene delivery/transfection is the key to CAR-T preparation, and the efficiency of vector introduction affects the cost and quality of CAR-T products. The advantages of selecting lentiviral vectors for CAR-T cell production include, among others, the following:

More effective and extensive infection of dividing and non-dividing cells and tissues;
Greater packaging capacity;
Lower cytotoxicity and immunogenicity;
Better expression effects.

References:

Poorebrahim, M.; et al. Production of CAR T-cells by GMP-grade lentiviral vectors: latest advances and future prospects. Crit Rev Clin Lab Sci. 2019 Sep;56(6):393-419.
Vormittag, P.; et al. A guide to manufacturing CAR T cell therapies. Curr Opin Biotechnol. 2018 Oct; 53:164-181.

* For research use only. Not intended for any clinical use.

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CAR-T Lentiviral Particles Helping to Establish an Efficient and Simple Platform for CAR-T Cell Expansion and Generation

CAR-T Process Overview

How to Generate CAR-T Cells using Lentiviral Vectors?

How to Achieve Efficient Transduction of CAR?

Benefits of Selecting Lentiviral Vectors for CAR-T Cell Transduction

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