KACTUS Launches CD3-TCR Complex VLP Products

T cells are the executors of the anti-tumor immune response. This is particularly true for cytotoxic CD8+ T cells, which are the primary force in the immune system for killing tumor cells. Enabling T cells to more effectively recognize and kill tumor cells is one of the core strategies of current immunotherapy (TCE, CAR-T, TCR-T, etc.).

The CD3-TCR Complex is the Molecular Hub for T-cell Immune Recognition and Activation

The CD3-TCR complex (T-cell receptor-CD3 complex) is a multi-subunit membrane protein complex composed of the antigen recognition module TCR (αβ or γδ), and the signal transduction modules CD3γε, CD3δε, and ζζ homodimer. While the TCR itself is responsible for recognizing the antigen-MHC complex (pMHC), its cytoplasmic tail does not contain signaling structures. Instead, the signal transduction from TCR relies on the ITAM (Immunoreceptor Tyrosine-based Activation Motif) on the CD3 subunits. Consequently, the CD3-TCR complex serves as the fundamental engine of the adaptive immune system, where the TCR governs antigen recognition and the CD3 subunits drive cellular activation, ensuring robust downstream signal transmission.

CD3-TCR complex and its functional signal transduction motifs[1]

The Importance of the CD3-TCR Complex in Antibody Discovery

On the T-cell membrane, CD3 does not exist in isolation. It must be integrated within the entire CD3-TCR complex to form a stable transmembrane structure. The integrity of each subunit is highly dependent on precise transmembrane pairing and complex organization. Consequently, immunotherapies that utilize T cells as effectors (such as TCR-T, CAR-T, TCE, ICIs, etc.) rely on modulating or harnessing the signaling of this complex at various levels.

When CD3 subunits are expressed individually, their conformation, surface epitopes, and glycosylation patterns often deviate from their native state. In vivo, CD3 functions as part of the TCR–CD3 complex, where key regions are masked through interactions with the TCR. When expressed in isolation, these normally buried epitopes become artificially exposed, leading to off-target antibody responses. As a result, many antibodies generated using recombinant CD3 proteins fail to recognize the physiological complex and exhibit poor binding efficacy on native T cells.

For antibody discovery, particularly in developing T-cell-activating therapies like TCEs, the primary challenge is modulating the activation threshold and kinetic profile of the CD3-TCR complex through precise epitope selection and conformational control. Utilizing the full TCR-CD3 complex (or its cell-surface expression model) for drug design and affinity optimization is far more effective for ensuring functional accuracy and safety. This approach helps mitigate off-target effects and better control CD3 cross-linking–induced T-cell activation, which may contribute to reducing the risk of CRS (cytokine release syndrome) while maintaining anti-tumor activity.

Evolution of TCE design and its representative therapeutic indices[2] Dashed line (left figure): Total cytokine responses; Solid line (left figure): Efficacy response

KACTUS CD3-TCR Complex VLP Products

Following the development of CD3-TCR Complex Nanodisc products, KACTUS has leveraged its mature and high-efficiency recombinant protein expression platform to launch the brand-new CD3-TCR Complex VLP series. Featuring full-length protein sequences, these products more accurately replicate the native assembly of the three CD3 dimers and their physiological association with the TCR. They are ideal for diverse applications, including animal immunization, antibody screening, epitope mapping, structure-guided design, and in vitro or in vivo validation. Experimental results confirm that KACTUS’s CD3-TCR complex VLP maintains dual biological binding activity, interacting effectively with both CD3 antibodies and their corresponding pMHC complexes.

Data Examples:

Immobilized Human NY-ESO-1 TCR&CD3 Complex VLP at 5 μg/mL (100 μl/well) on the plate. Dose response curve for OKT3, mFc Tag with the EC50 of 3.6 ng/ml determined by ELISA (QC test).

Immobilized Human NY-ESO-1 TCR&CD3 Complex VLP at 5 μg/mL (100 μl/well) on the plate. Dose response curve for Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) Monomer, His Tag (Cat. MHC-HM405) with the EC50 of 7.5 ng/ml determined by ELISA.

Furthermore, while screening with in vitro recombinant CD3 proteins has inherent limitations in predicting in vivo performance, these proteins remain valuable for generating a diverse initial antibody pool during early-stage discovery. This diversity enables more effective downstream optimization through cell-based or functional assays. Additionally, they are essential for structural and biochemical analyses, such as affinity quantification, competitive binding assays, and conformational studies. All of which are critical for selecting a CD3 arm with the optimal balance of affinity and structural integrity. To support these varied research needs, KACTUS provides CD3 monomers and heterodimers alongside the full CD3-TCR complex (VLP & Nanodisc).

Product List

References

[1] From TCR fundamental research to innovative chimeric antigen receptor design. Nat Rev Immunol. 2025 Mar;25(3):212-224. doi: 10.1038/s41577-024-01093-7.

[2] Innovative strategies for T cell engagers for cancer immunotherapy. MAbs. 2025 Jul 10;17(1):2531223. doi: 10.1080/19420862.2025.2531223.