Precision BioScience 在2018年三月份Best Practice & Research Clinical Haematology上發表了其off-the-Shelf T細胞治療技術體系Off the shelf T cell therapies for hematologic malignancies。
Off the shelf T cell技術流程:文章系統介紹了Off the shelf T cell技術
1. Gene editing for production of allogeneic CAR-T cells from unrelated healthy donor cells
2. Allogeneic CAR-T cells with targeted CAR insertion by homologous recombination
3. Additional gene edits to reduce immunogenicity and resist suppression within the tumor microenvironment
4. Allogeneic CAR-T cell manufacturing
5. Allogeneic CAR-T cell analytical methods
5.1 Viability and cell count
5.2 Cellular phenotype
5.3 Functional analysis
Traditionally, functional response of any T cell to antigenic
challenges is estimated by cytokine production, cytotoxic activity, and T
cell proliferation. Interferon gamma is a classical target to analyze T
cell potency and is utilized as indicative of the biological
functionality of manufactured T cell products. Conventional ELISA is a
time-consuming procedure which requires well trained personnel or
additional expenses for the automatization of the entire procedure. The
Microfluidic Simple Plex cartridge-based assay on the ELLA instrument
(ProteinSimple, San Jose, CA) may be a viable alternative to traditional
ELISA. The entire procedure is handsfree and take approximately 90 min
for readout.