Performance of Evercode™ TCR v4 in Antigen-Stimulated Human T Cells

Key Takeaways:

• Robust TCR detection in antigen-stimulated human T cells
• Recovery of paired alpha and beta chain CDR3 sequences from single cells
• Consistent paired TCR recovery across virally-expanded samples

Experimental Design:

Single cell T cell receptor profiling enables high-resolution characterization of T cell clonotypes, including recovery of alpha and beta chain CDR3 sequences from individual cells.

To evaluate the performance of Evercode TCR v4, cryopreserved purified antigen-stimulated human T cells were processed using the Evercode TCR v4, Human VDJ + WT workflow. Samples included T cells reactive to CMV, HSV, and Flu antigens, including HLA-restricted CMV-reactive and Flu-reactive T cell samples.

Analysis of the TCR repertoire demonstrated robust receptor detection, with TCRs identified in greater than 90% of T cells. Among cells with detected TCR sequences, 72% had paired alpha and beta chain CDR3 recovery.

Figure 1. TCR Detection and Paired CDR3 Recovery with Evercode TCR v4. UMAP visualization showing cells with detected TCR sequences, alongside quantification of paired alpha and beta chain CDR3 recovery. TCRs were detected in greater than 90% of T cells, with 72% of TCR-positive cells having paired TCR alpha and beta CDR3 recovery.

Paired alpha and beta chain recovery was observed across antigen-stimulated T cell samples. Most samples had paired CDR3 recovery above 70%, including CMV HLA-A0201, HSV gD1, and Influenza M1-reactive T cells. The CMV HLA-B35 sample showed lower paired recovery at 46%, while the full sample set ranged from 46% to 84%.

Figure 2. Paired CDR3 Recovery Across Antigen-Stimulated T Cell Samples. Bar plot showing the percentage of T cells with paired alpha and beta chain CDR3 recovery across CMV-, HSV-, and Flu-reactive T cell samples.

Together, these data demonstrate that Evercode TCR v4 supports robust single cell TCR profiling in antigen-stimulated human T cells, enabling detection of TCR sequences and recovery of paired alpha and beta chain CDR3s for clonotype analysis.

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