Chimeric Antigen Receptor T-cell Therapy, CAR-T, is a type of cancer immunotherapy that has gained world-wide interest in recent years, especially after FDA approval of CAR-T treatments for B-cell acute lymphoblastic leukemia and large B-cell non-Hodgkin’s lymphoma.  CAR-T therapy utilizes a genetic modification of a patient’s T-cells to express a target that is specific for the tumor antigen. The modified T-cells are expanded in the lab then infused back into the patient to target and kill the tumor cells. Treatment with this therapy has resulted in complete remission in some patients and the modified T-cells are persistent.

To ensure the safety of the CAR-T treatment, regulatory agencies require thorough characterization of the manufactured CAR T-Cells.  In addition, during the clinical trial or patient treatment phase, the level of modified T-cells in patients’ blood must be monitored frequently in a timely manner.

Examples of Genomic Tests for CAR-T Development

Copy Number Analysis for CAR T-cells

Regulatory agencies require CAR-T sponsors to determine the copy number of the inserted genes in manufactured CAR-T cells before releasing for patient treatment.

QPCR for Preclinical Biodistribution Study

In vivo toxicity studies, and sometimes shedding study, are required by regulatory agencies to evaluate the safety of therapeutic cells. QPCR, ddPCR, and NGS amplicon sequencing methods can be used to accurately assess the quantity of therapeutic cells in animal blood, CFS, and tissues for biodistribution studies.

Integration Site Analysis

NGS, PCR, and Sanger sequencing methods are used to delineate and validate potential insertion junctions and their locations in host genome.

Replication Competent Lentivirus Assay for CAR T-cells

Lentivirus is commonly used to generate CAR-T cells due to its ability to insert genes into host genome.  It is critical to ensure that there is no replication competent lentivirus (RCL) in the therapeutic cells.  This can be accomplished using QPCR and ddPCR methods.

Testing of RNAs in Animal Tissues and Human Blood

The study of mRNA or miRNA in animal tissues and in human blood via QPCR and ddPCR assays for both relative and absolute quantification of RNAs.

Avance Biosciences is specialized in assay development, assay validation, and sample testing using DNA sequencing, Next-Gen sequencing, QPCR, ddPCR, and many other biological services in compliance with GLP or cGMP regulations.  We are a leader in providing high quality genomic tests to support our clients in their CAR-T therapy development efforts.