Production of lentiviral vectors in scalable conditions for generation of CAR-T cells for cell therapy

• Grant number: 18/13307-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: November 01, 2018
• End date: September 30, 2020
• Field of knowledge: Engineering - Chemical Engineering
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Kamilla Swiech Antonietto
• Grantee: Mariane Cariati Tirapelle
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 14/50947-7 - INCT 2014: in Stem Cell and Cell Therapy, AP.TEM

Abstract

Studies with the use of modified T lymphocytes with Chemical Antigen Receptors (CAR) present expressive results in the treatment of leukemias and lymphomas. Lentivirus are widely used with vectors of gene modification in this type of therapy, with the reason of their advantages, such as cell genome stability, transduction efficiency and safety. Monolayer cultivation is limited because it does not respond to the demand for large-scale prodution of these vectors, requiring suspension cell culture for application in biorreactor. The aim of this study is to perform the production of lentiviral particle in scalable conditions and according to Good Manufacturing Practices, using sérum-free media in suspension culture for a generation of CAR-T cells. Therefore, HEK293T cells adapted to serum-free suspension culture will be used to produce lentivirals particles expressing a synthetic anti-CD19 protein. Initially, small-scale experiments will be performed by transient transfection using PEI. In this step, parameters such as, use of additives to increase efficiency and ratio of plasmids and PEI transfection agent, will be evaluated and applicated in the production in stirred tank bioreactor. With this Project we hope to develop an efficient, reproducible and economically viable platform for the production of lentiviral vectors that contributes significantly to the research related to cell immunotherapy.

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

TIRAPELLE, Mariane Cariati. Production of lentiviral vectors under scalable conditions for the generation of CAR-T cells for cell therapy. 2021. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.