TD2 Oncology Expands Flow Cytometry Expertise to Advance In Vivo CAR T Therapies for Autoimmune and Oncology Development

In response to advances in In Vivo CAR-T engineering, TD2 Oncology launches
flow cytometry panels to track CAR-T persistence, activation, and efficacy.

Scottsdale, AZ – TD2 Oncology, a leader in advanced analytical services for cell and gene therapies, today announced expanded capabilities in flow cytometry assays tailored for In Vivo CAR-T treatments. This service supports researchers and developers in preclinical models and clinical trials by providing high-resolution monitoring of CAR-T cell dynamics, addressing key challenges like persistence and off-target effects highlighted in recent scientific literature. As in vivo CAR-T emerges as a scalable alternative to ex vivo manufacturing, TD2 Oncology's assays enable faster, more reliable data to advance therapies for autoimmune diseases, hematological malignancies, and solid cancers.

Growth of In Vivo CAR-T

In vivo chimeric antigen receptor (CAR) cell therapies represent a paradigm shift from traditional ex vivo CAR-T approaches, where patient T cells are extracted, engineered, and reinfused. By using targeted delivery systems like lipid nanoparticles (LNPs) or viral vectors to introduce CAR-encoding genetic material (e.g., mRNA or DNA) directly into the patient's immune cells, these therapies eliminate manufacturing complexities, reduce costs, and avoid lymphodepleting chemotherapy. In vivo CARs demonstrate promise for autoimmune diseases, where autoreactive B cells drive pathogenesis in conditions like systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and multiple sclerosis (MS) as well as in patients with relapsed B-NHL patients, HER2+ breast and gastric cancers, and renal cell carcinoma. Preclinical and early clinical data demonstrate efficient CAR expression, B-cell depletion, immune reset, and clinical remission with minimal toxicity including cytokine release syndrome and neurotoxicity.

Importance of Robust Analytical Monitoring

Success for In Vivo CAR-T hinges on real-time assessment of cell persistence, activation, and tumor targeting–parameters where flow cytometry excels. Preclinical and clinical studies rely on multiparametric flow assays to quantify CAR-T expansion (e.g., via CD4/CD8 subsets and exhaustion markers like PD-1), cytokine profiles (e.g., IFN-γ, IL-2), and target cell depletion (e.g., CD19+ B cells). Flow cytometry panels can show good sensitivity in detecting CAR-T at low levels of viable T cells, which can then be used in correlating therapeutic outcomes in models like NSG mice engrafted with human lymphoma tumors. These assays outperform qPCR by providing phenotypic insights, for example, granzyme B expression for cytotoxicity, essential for predicting relapse in relapsed/refractory diffuse large B-cell lymphoma (R/R-DLBCL).

TD2 Oncology’s Expanded Flow Cytometry Platform

TD2 Oncology's flow cytometry platform offers end-to-end support for In Vivo CAR-T studies, from assay design to data interpretation. Key features include:

• Preclinical Optimization: Longitudinal tracking in xenograft models using low-volume blood samples (as little as 50 µL) with custom flow cytometry panels to monitor CAR+ T-cell persistence, proliferation, focused mainly on the memory differentiation and activation/exhaustion status of expanded CAR+ T cells, endogenous immune populations, and malignant cell levels.
  
  
• Clinical Monitoring: Real-time phenotyping post-infusion, including CAR detection via the anti-CAR detection antibody. Sampling schedules must be optimized to capture expansion, peak, and contraction phases of CART kinetics, and data integration should link cellular phenotypes with clinical endpoints to enable meaningful pharmacokinetic and pharmacodynamic interpretation.
  
  
• Assay Considerations: In Vivo CART flow cytometry assays require careful alignment of specimen selection with matrix specific constraints, as whole blood, bone marrow aspirates, PBMCs, and tissue dissociates each present unique challenges such as debris, autofluorescence, or cryopreservation effects. Detection sensitivity hinges on the choice of reagents, with high affinity, low background anti-CAR antibodies being essential for reliable quantification. Comprehensive panel design should incorporate lineage, activation, memory, exhaustion, and trafficking markers to capture both CAR+ T cells and endogenous immune populations. Spectral or higher dimensional flow cytometry offers advantages in resolving dim signals and complex matrices, while robust gating strategies, appropriate controls, and standardized workflows ensure reproducibility across studies.

TD2 Oncology also provides customized panel development and fit-for-purpose assay validation aligned with each therapy mechanism of action. Flow cytometry data can be integrated with qPCR and other orthogonal methods to deliver a full biodistribution profile that supports IND enabling studies and clinical trials. "In Vivo CAR-T therapies represent a significant advance toward off the shelf therapies for patients, and development success depends on precise analytical tools that can capture complex and variable responses in real time,” said Dr. Jennifer Stewart, VP, Flow Cytometry Strategy at TD2 Oncology. “Our comprehensive flow cytometry services enable developers of in vivo CAR for autoimmune or oncology applications to refine next generation constructs such as nanoparticle delivered mRNA CARs, supporting the translation from preclinical proof of concept to early human studies."

Industry Momentum

Big Pharma acquisitions signal confidence: AbbVie ($2.1B for Capstan's LNP CAR-T in lupus, 2025), Eli Lilly ($1B for Verve's in vivo platform), and Gilead ($~300M for Interius' CAR for autoimmunity) total >$5B in 2025 deals. As of December 2025, ClinicalTrials.gov lists approximately 60 registered clinical trials specifically for in vivo CAR (chimeric antigen receptor) cell therapies targeting autoimmune diseases and cancer combined with >20 new in vivo CAR registrations in 2025 alone.