UCL CAR T GMP manufacturing team

The UCL academic CAR T cell program is the most comprehensive of its kind in Europe. Our program encompasses a broad-ranging portfolio of studies actively recruiting patients with B and T cell malignancies, myeloma, and neuroblastoma. We have several new trials in the pipeline to target midline glioma, glioblastoma and Hodgkin’s lymphoma, to name just a few. 

CAR T manufacturing

Our role in the UCL CAR T GMP manufacturing team is to translate early research finding into clinical products and deliver new CAR T cell therapies for patients enrolled in the Phase 1 academic CAR T trials. We also focus on the development of improved manufacturing methods and new technologies.

Contact email: rfh_pulelabs@live.ucl.ac.uk.

What is GMP?

Good Manufacturing Practice (GMP) describes a minimum set of standards which a manufacturer must meet in order to complete their processes. In our case, it defines a set of standards which we must achieve to be able to manufacture Advanced Therapeutic Medicinal Products (ATMP). Facilities and teams producing ATMP are regularly inspected by the Medicines & Healthcare products Regulatory Agency (MHRA) who verify that therapeutic products are of consistently high quality, are appropriate to their intended use and that they meet the requirements of the product specification. The MHRA can remove licenses to groups manufacturing ATMP if the criteria are not met, or where manufacturing facilities do not meet required specifications. 

Once all preliminary work for a new trial has been completed, various regulatory paperwork and data is submitted to the MHRA, who will ultimately decide whether the trial can be opened. The same applies to substantial amendments for existing trials, for example a major change to a protocol, requests for trial extensions to include additional disease targets, patient eligibility criteria or changes to the release criteria for an ATMP.  

Meet the team

Juliana Dias, PhD
Head of GMP CAR T Manufacturing 

Juliana graduated from Federal University of Rio de Janeiro with a BSc in Biophysics/Biotechnology, then completed her PhD in Biophysics at the same university. She worked in clinical cell therapy for six years and joined UCL in 2017 to set up the UCL CAR T manufacturing platform at the CCGTT. Currently, she leads the CAR T manufacturing team for the development and delivery of new products for the UCL Phase 1 clinical trials.

Giulia Agliardi, PhD 
Senior Research Fellow  

Giulia graduated from University San Raffaele in Milan (Italy) with a degree in Medical Biotechnologies. She holds a PhD from UCL, where she developed CAR T cell approaches for solid tumours, in particular high-grade gliomas. Giulia joined the GMP CAR T cell manufacturing team in 2021, where she leads Quality Control activities for all trial manufactures, as well as process development and optimisation work for introduction of upcoming Phase I trials. 

John Garcia, PhD
Senior Research Fellow

John holds a BSc in Cell and Molecular Biology (Université de Lorraine, France), an MSc in Cell and Tissue Engineering (Keele University) and a PhD in Bioengineering and Regenerative Medicine (Keele University). With 10 years of experience in cell therapies, John contributed to clinical trials for cartilage regeneration at the RJAH Orthopaedic Hospital before joining the GMP CAR T cell manufacturing team in 2020. John’s current role involves technologie transfer, initiating new trials and process optimisation.

Louisa Green, MSc
Senior Scientist (Cleanroom Lead)

Louisa has been working in the field of T cells since 2017 and holds a Master's in Immunology from Imperial College London. Within the Pule team, she is responsible for training new members of staff in cleanroom activities, and overseeing the cleanroom manufacturing process. Louisa also works to review all batch manufacturing paperwork ahead of product release for each patient. 

Mhairi Vaughan, MSc
Senior Scientist (QC Lead)

Mhairi’s role is focused on Quality Control, using cell characterization techniques such as multiparametric flow cytometry for product release testing. She is also responsible for carrying out process development activities, training junior QC staff, and assisting in cleanroom activities. She holds a BSc in Biology from the University of Bristol and completed her Master’s at the London School of Hygiene and Tropical Medicine (LSHTM) before joining the RFH team in 2019.

Evie Lewin, MSc
GMP Production Technician

Having progressed within the team from an administrative role, Evie currently works with Louisa in the cleanrooms, supporting the main manufacturing activities. Evie has brilliant attention to detail, and can be relied upon to make sure all documentation prior to submission is correct.

Laís Castro, BSc
GMP Production Technician

Lais' role is focused on assisting with the in-process and QC of ATMPs for clinical trial use, and contributing with process developmental activities.

Laís is also responsivle for preparing QC reports and stock control of materials and reagents. 

Ellie Wooderson, BSc
Laboratory Technician

Ellie prepares batch-related paperwork ahead of manufacturing and maintains laboratory and patient records to GMP standards. She is also responsible for managing stock levels for the manufacturing team and prepares shipment of ATMP to their respective sites.

Silvia Rondinelli, MSc
Laboratory Technician 

Silvia graduated from University of East London with a BSc in Biomedical Science and then completed her Masters in Neuroscience and Translational Medicine at Queen Mary University of London. Her role involves assisting in the development of potency testing assays, as well as contributing to Process Development activities for different trials. She also performs flow cytometry analysis of CAR T products and assists in the quality control of viral vectors. 
 

Laboratories - process development

Preliminary and scale-up work for new trials is performed in our Process Development (PD) laboratory. Here, research-grade reagents are used to optimize and fine-tune manufacturing protocols, without the constraints of a cleanroom environment.

We have access to two CliniMACS Prodigies in this laboratory to perform PD-level manufactures, prior to cleanroom validation runs.

In a shared QC lab, we have access to a flow cytometer, which is used for analysis of both GMP and PD samples

Laboratories - cleanrooms

Cleanroom - Royal Free Hospital

Once ready for Phase 1 clinical trial, our work is conducted in a specialized laboratory called a cleanroom. Use of the cleanrooms is tightly regulated, with users required to don a significant amount of PPE upon entry including gowns with hoods, goggles and boots. The cleanroom facility at the Royal Free Hospital is large and comprises five grade B, one grade C and four grade D laboratories.

All consumables and equipment are sterilized with cleanroom-grade reagents prior to transfer, and during procedures the environment is monitored via agar contact and settle plates, as well as particle counters within the microbiological safety cabinets. At the end of each session, the rooms and equipment are cleaned again to avoid contamination for the next team. 

The Pule team has exclusive use of two CliniMACS Prodigies in the C-grade laboratory. This enables us to manufacture two products at the same time, which shortens waiting times for patients.

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Typical workflow from idea to manufacture:

1. Binder discovery

• Identification of target antigens
• Development and selection of specific binders

2. Construct optimisation

• Selecton of CAR designs
• Viral vector production
• Inclusion of additional features (e.g., suicide/marker genes)

3. Pre-clinical validation

• Small-scale assessment of CAR T function
• Use of lab models to evaluate capacity to control tumour burden
• If applicable, validation of genome editing strategy

4. GMP vector production

• Manufacture of selected viral vector to GMP compliance

5. Method optimisation

• Small-scall optimisation using GMP reagents
• Definition of culture conditions and timelines for GMP process

6. Scale-ups / engineering

• Method adapted for large-scale use
• Definition of QC testing plan and methods
• x3 GMP cleanroom validation runs
• Validation of testing and manufacturing method

7. Regulatory submission

• All data, proposed trial protocol, product dossier and associated documents are prepared and submitted to the MHRA and for ethics approval

8. Trial initiation

• Product manufacture as per validated and approved protocols
• Batches are reviewed and certified by a Qualified Person
• All activities carried out within a Quality Management system
• Continuous reviw of processes and trial results

Stages of ATMP manufacture

Once all pre-clinical work and the regulatory submission processes are complete, patient recruitment can begin.  Patients must meet a series of eligibility criteria including previous unsuccessful conventional treatment attempts, as well as undergo a thorough screening process for several infectious disease markers. 

Once recruited for a trial, the patient undergoes leukapheresis apheresis, which is shipped to the manufacturing site on the same day. The 8-day automated ATMP manufacture process can now begin. Once the product has been successfully cryopreserved, the departmental Qualified Person (QP) verifies all manufacture paperwork and sterility testing results. Any non-conformances or related risk assessments are also reviewed. The product can then be certified and shipped back to the hospital for infusion.

Our trials

COBALT:

Evaluation of CAR19 T-cells as an optimal bridge to allogeneic transplantation. This trial is designed to test the feasibility and safety of CAR19 therapy as a bridge to transplant in patients with DLBCL relapsed after auto-HSCT, or refractory to primary salvage. Three weight-dependant doses are made for each patient.   

• Status: Completed

CARD:

CAR19 Donor Lymphocytes for relapsed CD19+ malignancies following allogeneic transplantation. This trial uses matched healthy donor starting material, as opposed to the patient’s own cells.   

• Status: Completed    

CARPALL:

Immunotherapy with CD19 CAR redirected T-cells for high risk/relapsed paediatric CD19+ acute lymphoblastic leukaemia and other haematological malignancies. This paediatric trial is still ongoing, but manufacture is now taking place at Great Ormand Street Hospital (GOSH).  

• Status: Completed 

CAROUSEL:

Immunotherapy using CAR T-cells to target CD19 for relapsed/refractory CD19+ Primary Central Nervous System Lymphoma. Two weight-independent doses are made, although infusion of the second dose (through an Ommaya reservoir directly into the brain) is dependent on tolerability of the first intravenous dose. 

• Status: Completed

ALLCAR19:

Immunotherapy for high risk/relapsed CD19+ Acute Lymphoblastic Leukaemia, B-cell Non-Hodgkin’s Lymphoma (BNHL) and Chronic Lymphocytic Leukaemia (CLL)/ Small Lymphocytic Lymphoma (SLL) using CAR T-cells to target CD19. This trial initially opened to ALL patients only but due to its success, an extension was granted for a further 30 patients with NHL, CLL and SLL. ALLCAR19 is our most successful trial to date, and the ALL branch is now in Phase II. 

• Status: Open/recruiting 

MCARTY:

Phase 1 study evaluating the activity of Modular CAR T for myeloma. This trial opened in 2022, so is still in the preliminary stages. Depending of the success of the first cohort of patients, the trial will progress to the second cohort where participants will receive duel D8/CAT19 transduced autologous CAR T cells

• Status: Open/recruiting 

KCAT19:

Allogeneic T cells expressing T cell receptor-KDEL and the chimeric antigen receptor CAT19 for the treatment of advanced CD19+ malignancies. This trial uses cord blood from healthy donors and is open to patients with high-risk relapsed/refractory B cell malignancies who are not eligible or suitable for the ALLCAR19 trial. As healthy donors are used, doses are frozen in universal batches with the ultimate aim being an ‘off-the-shelf’ service. The major advantage of this is that patients won’t have to wait for cells to be manufactured, sterility results to be returned and the subsequent QP release process (typically totaling 4-5 weeks).

• Status: Open/recruiting

Publications and news features

• Summary on the CAR T manufacturing process: Manufacturing chimeric antigen receptor T cells: issues and challenges 
• Dr Claire Roddie academic profile
• BBC documentary War in the Blood
• UCL CAR T programme

Contact us

Email: rfh_pulelabs@live.ucl.ac.uk