Clinical-grade CAR T cell manufacturing

1. Separation of T cells

Cell separation on the CliniMACS Prodigy is a simple and easy method to magnetically isolate T cells with a high purity. Advantages of this completely automated method include:

• Efficient separation of CD4⁺ and CD8⁺ cells without inducing activation

• Removal of contaminating immune cells to enhance T cell transduction efficiency

• No magnetic bead removal necessary

2. T cell activation

MACS® GMP T Cell TransAct™ is a colloidal polymeric nanomatrix conjugated to humanized, recombinant CD3 and CD28 agonists. It is a ready-to-use reagent that has been designed to activate and expand enriched T cell populations or resting human T cells from peripheral blood mononuclear cells (PBMCs). Thanks to its nanoscale structure the reagent can be sterile filtered, and excess reagent can conveniently be removed by washing the TransAct activated cells.

MACS GMP T Cell TransAct is directly applicable for the use in automated culture systems, such as the CliniMACS Prodigy Instrument.

3. Lentiviral transduction

To achieve long-term gene expression through stable genomic insertion, two types of gene transduction tools are commonly used: gamma-retroviral vectors (RV) or lentiviral vectors (LV). The CliniMACS Prodigy TCT Process supports the use of both vectors.

For easy integration of a transduction enhancer for RV-based transduction, Vectofusin-1® can enhance your RV transduction efficiency. Vectofusin-1 is compatible with both static and spinoculation protocols. LV can easily be integrated into the automated CliniMACS Prodigy TCT Process. We also deliver tailored LV solutions from a strong expertise in advancing LV design and technologies that meets global GMP requirements.

4. CAR T cell expansion

Cell expansion takes place in the cell cultivation chamber, which is part of the CliniMACS Prodigy Tubing Set in order to maintain a functionally closed system. As part of the TCT Process activity matrix, the culture duration as well as the medium supply and exchange are flexibly programmable.

Ideal cultivation and expansion of transduced T cells rely on the strong synergy of MACS GMP T Cell TransAct™, TexMACS™ GMP Medium, and MACS GMP Cytokines.

TexMACS GMP Medium

• Specifically developed for T cell cultivation

• Serum- and xeno-component free

• Pharmaceutical-grade human serum albumin

• QC functionality test on every batch

MACS GMP Cytokines

• Lot-to-lot consistency and lot-specific certificates of analysis

• Designed according to the recommendations

of <USP 1043> on ancillary materials

• Manufactured and tested under a certified ISO 13485

quality system

In-process and quality control and clinical flow cytometry

In-process control and quality control (IPC/QC) are required for consistent cell manufacturing. Integrated sampling pouches on the CliniMACS Tubing Sets allow for controls to be collected at any time during, and at the end of the cell manufacturing process. Miltenyi Biotec provides complete solutions with its MACS Flow Cytometers, a wide portfolio of MACS IVD Antibodies, and automated flow cytometry analysis with the CAR T Cell Express Mode Package for clinical flow cytometry. MACS IVD Antibodies are produced according to the quality management system ISO 13485.

When using a MACSQuant® Instrument, Express Modes allow you to fully automate and standardize your flow cytometry process. The MACSQuant Instrument and the MACS Flow Cytometry portfolio are for research use only.

Disclaimer

^{For the manufacture and use of gene-engineered T cells, national and international legislation and regulations must be followed. The manufacturer has to validate the QC method. Miltenyi Biotec as the provider of products for the T Cell Transduction Process does not give any recommendation regarding the use of the manufactured cells for therapeutic purposes and does not make any claim regarding a clinical benefit.}