Rhesus macaques are a popular animal model in biomedical research, providing valuable data for evaluating the safety and efficacy of vaccine and drug candidates in clinical trials. In HIV research, understanding the immune response of these non-human primates to vaccination is crucial in optimizing vaccine strategy and assessing safety.

The first solution of its kind, the SMART-Seq Human TCR (with UMIs) allows scientists to easily capture TCR sequences from rhesus macaque RNA and analyze the data with our accompanying bioinformatics tool, Cogent NGS Immune Profiler Software (CogentIP).

In this internal study, we tested 10 ng (RM-10) and 100 ng (RM-100) of total RNA isolated from PBMCs of three rhesus macaques, spanning the high and low ends of the input range as the protocol recommends, and libraries were generated following this kit’s workflow. Using our kit, we can capture a good number of clonotypes from both TCR alpha and beta chains, and the clonotype counts consistently increased with the increasing amount of RNA input (Panel A). Meanwhile, the kit shows outstanding reproducibility when being used on rhesus macaque samples, with Pearson correlation scores exceeding 0.9 for both TRA and TRB libraries prepared with various input amounts (Panel B and C).

SMART-Seq Human TCR (with UMIs) sequencing libraries from rhesus macaque samples show highly reproducible clonotype detection. RM-10 and RM-100 libraries were created in triplicate from PBMC of three rhesus macaque donors. Full-length TRA and TRB sequences were obtained using the Illumina MiSeq® platform. Average reads/sample were 3 x 10⁶ for RM-10 and 1 x 10⁶ for RM-100. Clonotypes were identified using CogentIP. The total number of clonotypes per input amount and gene target were identified in one representative donor (Panel A). Error bars represent the standard deviation among triplicates. Two replicates from a representative donor's RM-10 (Panel B) and RM-100 (Panel C) libraries were chosen for pairwise comparisons of overlapping TRA and TRB clonotypes. Scatterplots compare fraction of total reads assigned to common clonotypes between replicates.

SMART-Seq Human TCR (with UMIs)

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