Full-Length TCR/BCR Repertoire Profiling

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Full-Length TCR/BCR Repertoire Profiling

CD Genomics provides reliable and comprehensive TCR/BCR repertoire analysis for blood, bone marrow, or tissue samples based on long-read sequencing technology, PacBio SMRT sequencing. We can guarantee the consistency of the sequencing process and analysis results for each sample throughout the experiment, and perform detailed bioinformatics processing and data analysis of the sequenced data. Our service is accurate and sensitive in requiring little DNA or RNA input, is well suited for immunization projects requiring high-precision sequences >600 bp to several kb.

Overview of TCR and BCR

T lymphocytes (T cells) and B lymphocytes (B cells) are the main cells responsible for the adaptive immune response and their antigen recognition depends mainly on the T cell receptors (TCRs) and the B cell receptors (BCRs). These two types of cell surface molecules share a common characteristic of being very diverse, thus capable of recognizing a wide range of antigenic molecules. TCR heterodimer mostly consists of two subunits, the α and β chains and to a low percentage of γ and δ chains, each the product of a different germline TRATRBTRG, or TRD gene locus, respectively.  BCR is a heterotetramer consisting of two heavy chains of membrane immunoglobulins encoded by the IGH loci and two light chains of immunoglobulin kappa or lambda encoded by the IGK or IGL genes. Notably, each of these gene loci has gene segments in their germline configuration, namely variable (V), diversity (D), and joining (J) genes. During T or B lymphocyte development, one member of each cluster is joined together by irreversible somatic DNA rearrangement, a process also called V(D)J recombination. Further intercellular diversity is produced by the random addition or removal of nucleotides on V(D)J junctions, which encode the complementary determining region 3 (CDR3) of the receptor antigen-binding site. The immunome repertoire, especially the TCR profile, changes considerably under different disease conditions, which is an important guide for diagnosis and treatment.

Our Service Workflow

Our service is based on the PacBio Sequel II System and can analyze full-length TCR/BCR genes, providing more information to facilitate distinguishing unique variants. Accurate sequences are also ensured to confidently identify specific clonotypes. We can not only obtain information about the expression of different T-cells in the population from genomic DNA but also directly observe the functional expression of genes, starting from mRNA. In addition, our bioinformatics pipeline for immune repertoire analysis is continuously optimized to provide accurate insights into the diversity, dynamics,and clonality of immune repertoire.

Our Service Workflow.-CD Genomics

Sample Requirements

  1. Multiple acceptable sample types, including sorted cells, PBMCs, whole blood, bone marrow, fresh frozen tissue, and so on.
  2. Species: human.
  3. Target: DNA or RNA.
  4. Sequencing technology: the PacBio Sequel II System.

Bioinformatics Analysis

Raw sequencing data are automatically processed to facilitate immediate genetic evaluation, including demultiplexing, and generation of adapter trimmed FASTQ files.

  • Reconstruction of TCR/BCR clones.
  • Complementarity determining region 3(CDR3) identification.
  • Clonality and diversity estimation.
  • Advanced bioinformatic options are available on request.

Application of Our Services

  • Insights into the development, functions, and diversification of T and B cells.
  • Discovery of biomarkers that have the potential to predict future immunotherapies.
  • Monitoring and evaluating drug therapy efficacy, such as immunotherapies in various cancers.
  • Quantify somatic cell hypermutation to potentially investigate malignant lymphocyte transformation.

Please don’t hesitate to contact us. Discuss your projects with our professional scientists.

References

  1. Omer, A., et al. (2022). "T cell receptor beta germline variability is revealed by inference from repertoire data.” Genome medicine, 14(1), 1-19.
  2. Singh, M., et al. (2019). “High-throughput targeted long-read single cell sequencing reveals the clonal and transcriptional landscape of lymphocytes." Nature communications, 10(1), 1-13.
For Research Use Only. Not for use in diagnostic procedures.

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