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Single-cell Immune Repertoire Sequencing

Single-cell Immune Repertoire Sequencing

The immune repertoire (Immune Repertoire, IR) is the detection of B/T lymphocyte antigen receptor diversity, that is, the variety of B-cell receptors (BCRs) and T-cell receptors (TCRs). BCR and TCR generally consist of a variable region (V region), a variable region (D region), a linker region (J region), and a constant region (C region). Meanwhile, the V(D)J region undergoes rearrangement or recombination in the genetic process, thus forming a large number of combinations and resulting in extremely rich sequence diversity for BCR and TCR genes. Together, these sets of sequence diversity are named the immune repertoire.

The single-cell immune repertoire sequencing (scVDJ-seq) technology can simultaneously detect gene expression by correlating transcriptomic and TCR/BCR diversity at the single-cell level, which can not only support a clear understanding of TCR/BCR clonotypes but also combine transcriptomic data to dig deep into the life mechanism. scVDJ-seq provides a more efficient platform for immunology research and has advanced application prospects in research fields, such as tumor microenvironment, infectious diseases, and autoimmune diseases.

As the scVDJ-seq technology can detect nearly 10,000 cells simultaneously in one single experiment, it allows the acquisition of mRNA expression profiles at the 5' end of transcripts as well as TCR and BCR information in one sample. Hence, combining gene expression profiles and V(D)J data can analyze the impact of immune response on complex tissue samples, monitor immunotherapy effects, and study the molecular mechanisms of onset and progressive diseases.

Our Single-cell TCR/BCR Sequencing Service

Our Immuno-profiling service couples ultra-deep sequencing of the V(D)J regions with customized bioinformatics, thus comprehending critical data on the physical cognate pairing of TCR or BCR chains within a single cell. In addition, recent advances in microfluidic technology allow the BCR and TCR chains to be paired at single-cell resolution.

Advantages of Our Single-cell TCR/BCR Sequencing Services

1. Extensive experience: Over 1,000 TCR/BCR completed projects and each project is committed to library construction of 1,000-1,0000 cells simultaneously while accessing the transcriptome and immunology data.

2. Strict quality control: Complete immune library sequencing and experimental quality control process to obtain the full-length V(D)J sequence pairing information.

3. Powerful bioinformatics analysis: Joint analysis of single-cell immune repertoire and gene expression to further elucidate cell interaction; personalized analysis available.

4. Double-chain precise matching: Efficiently match the heavy and light chains of BCR as well as the α and β chains of TCR at the single-cell level.

5. Single-cell expression: Simultaneous detection of full-length paired immune repertoire (TCR/BCR) and the 5' mRNA expression.

Bioinformatics Workflow

Single-cell Immune Repertoire Sequencing

Sample Requirements

Sample Type: tissue, blood, cultured cell line, prepared single-cell suspension.

Note: CD Genomics is willing to provide technical and experimental assistance with our greatest effort for tissue samples that can be dissociated to obtain a single-cell suspension. Due to the specificity of different sample types, we will not guarantee that the experimental method can apply suitably to all populations.

Quality Requirements:

Cell viability greater than 70%;

Concentration between 500-2,000 cells/μL;

Volume no less than 200 μL;

No cell culture medium and buffer should contain Ca2+ and Mg2+;

Total cell volume less than 40 μm.

Applications of Single-cell TCR/BCR Sequencing

1. Allow novel T/B cell exploration in terms of cell types and functions

2. Construct immune repertoire with more refined samples at the single-cell level

3. Accelerate antibody screening with single-cell immune repertoire compared to traditional B-cell antibody screening

4. Perform single-cell VDJ testing to aid in the development of tumor immunotherapies (TCR-T)

For research use only, not intended for any clinical use.

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