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In vitro kinase assay using purified enzyme played a fundamental role in developing variety of different kinase inhibitors for many years. Although testing inhibitor compound in this assay is usually robust depending on the assay conditions, it has obvious limitations that are difficult to improve; 1) activity of inhibitor is likely over-estimated because ATP concentration used in assay is lower than the one inside the cells (millimolar range), 2) it is not trivial to purify recombinant protein to have properties, like post-translational modification and so on, that are close to the enzyme produced inside the cells.
Ba/F3 kinase library has been constructed to overcome these limitations by conferring IL3-independent growth of Ba/F3 cells that are solely dependent on ectopically expressed, single protein kinase gene. By measuring growth inhibition when expressed kinase is inhibited by compound, the activity of inhibitor can be estimated inside the cells.
At Txinno Bioscience, Ba/F3 cell line library of 50 different kinases has been acquired and we are generating additional Ba/F3 cell lines for new target proteins (protein kinases or RAS protein) of interest to facilitate drug discovery.
Tumor is not a collection of simple cancerous cells, but rather a complex tissue composed of various types of cells under unique tumor microenvironment (TME). As our understanding of cancer is deepen, it becomes clear that interrogating cancer cell alone is not enough to fully understand various interactions between tumor cells and other types of cells in TME ( for example, an immunosuppressive environment established by tumor cells and immune cells). There has been steady progress in improving ex vivo cell culture system to mimic the TME in order to test various treatment moieties. However, it is remaining as a major hurdle to design an assay system that is 1) manageable enough to perform in conventional cell culture facility and 2) types of cells using and 3) flexible enough to test both human and mouse cell types.
In Txinno Bioscience, we developed two platform technologies to address these issues and facilitate our drug development process.
1) LIN (Lymphocyte Infiltration) assay system that can cultivate cancer cell spheroid and immune cells from PBMC in separate compartments of microplate wells. With immune cells appropriately stained for detection, the effect of drug treatment on infiltration of immune cells toward the spheroid can be measured.
2) TMED (Tumor MicroEnvironment in Dish) is a co-culture system that are composed of spheroid from cancer cell line, isolated human immune cells, cancer-associated fibroblast, adipocytes and fabricated extracellular matrix to closely mimic actual human TME.
LIN assay system developed by Txinno Bioscience can be a highly useful ex vivo culture system to measure the infiltration of immune by innate immune modulator reflecting conversion of TIL-deficient “cold tumor” to TIL-proficient “hot tumor” and to access the efficacy of IO drugs of various mode of action.
TMED system developed by Txinno Bioscience can be used to test various anti-cancer therapeutics, including chemical compound, biologics as well as cell therapy. Also, the system can be used to perform various genetic screening using genomic reagents, like CRISPR, to find the next generation cancer targets.
LIN & TMED system with human-derived cancer cells and immune cells (Version 1) was upgraded into a system can culture spheroid of cancer cells and CAF (cancer-associated fibroblast) with immune cells (Version 2). For next step, we are developing a co-culture system with cancer cells, CAFs, endothelial cells and extracellular matrix (Version 3), which closely mimic actual tumor microenvironment (TME) of patient tumors.