citations

                    Closed-system
transposon-mediated manufacture of GMP grade CAR T-cells via the Lonza
Nucleofector LV XL
                

Authors:

L.M. Brownrigg, S. Nichols, E. Bosio, B. Carnley, M. Sturm

In:

Source: Cytotherapy
Publication Date: (2020)
Issue: 22: S382

Research Area:

Immunotherapy / Hematology

Cells used in publication:

T cell, human peripheral blood unstim.: Species: human; Tissue Origin: blood

Platform:

4D-Nucleofector® LV-Unit

Experiment

The T-cells were washed, resuspended in P3 Nucleofector Solution and sequentially nucleofected with CAR19.41BBz transposon plasmid DNA in Nucleofector
LV XL cartridges. The cells were directly ejected immediately post-nucleofection into AIM V conditioned media for subsequent stimulation with gamma-irradiated antigen presenting cells and GRex culture expansion up to 15 days.

Abstract

Background & Aim A method for the non-viral transposon based manufacture of CAR19.41BBz T-cells for the Phase I CARTELL (UTN: U1111- 1204-6974) and AUTOCAR19 (U1111-1226-8682) clinical trials has previously been described. The original method was non-Good Manufacturing Practice (GMP) standard, and used a traditional opencontainer electroporation system. The method presented here is a modification of the original method to GMP standard to incorporate the fully enclosed Lonza 4-D Nucleofector LV Unit XL cartridge system and eliminate open-container cell handling and processing. For GMP applications, closed electroporation systems are preferable to open-container systems, such as the older 4-D X-unit nucleocuvettes offered by Lonza, as they reduce the risk of product contamination.

Closed-system transposon-mediated manufacture of GMP grade CAR T-cells via the Lonza Nucleofector LV XL - Cytotherapy (isct-cytotherapy.org)

Please see also more details in the thesis under the following link: THESIS_-_DOCTOR_OF_PHILOSOPHY_-_BROWNRIGG_Leon_Michael_-_2023_.pdf (uwa.edu.au)