Optimization of Lentiviral Vector Production at Research Scale

Abstract

Lentiviral vector is a tool that have been used in gene therapy to create CAR T-cell for the treatment of cancer, especially hematological malignancies. Event though it is not difficult to produce lentiviral vector, some of the steps in the process might lead to yield and functional loss. This research incorporates production of lentiviral vector using an adherent cell line, 293T cells. The process begins with the transformation of competent cells, purification of the plasmid, cell transfection, cell transduction, and the last is the detection of transgene expression. Out of the results, cells with 10 -2 dilution from 50,000 resulted in the highest yield of transduced cells with the value of 7.39 of GFP-A+ cells. This gives a viral titer of 739,000 TU/mL. However, the result for the successfully transduced cells were non-existent. This might be caused by several factors, such as low DNA vector quality, incorrect cell concentration or confluency, incorrect viral concentration, as well as human error.