Creative Biolabs offers the unparalleled mass sequencing service to analyze the immune repertoires for our customers all over the world. Depending on our cutting-edged SuPrecision™ platform which mainly based on next-generation sequencing (NGS) technique, our scientists have accumulated extensive experience in identifying the whole four types of T-cell receptor (TCR) chains (α, β, γ, and δ) and the vast majority of B-cell receptor (BCR) V(D)J sequences contained in a single sample.
The Properties of TCR and BCRFig.1 Structure, function, and diversification of TCR/BCR receptors (Lossius et al. 2016).
Cellular immune responses from T-cells and humoral immune responses from B-cells are stimulated by exposures to antigens, including pathogens, allergens, and neoantigens. V(D)J recombination in the primary lymphoid organs creates the incredibly diverse and unique repertoire of the hypervariable regions of BCR and TCR, and somatic hypermutations contribute to additional BCR diversity. During B and T-cell development, self-antigens are presented to B and T-cells to select out self-reacting types, and to ensure only B and T-cells that recognize and attack foreign antigens are in the circulation. T-cells only recognize foreign proteins presented on MHC, while B-cells can also target foreign DNA, lipids, or carbohydrates. Upon recognition of foreign antigens and with the presence of co-stimulatory molecules, B-cells and T-cells express cell surface activation markers, attack foreign antigens, secrete cytokines, stimulate each other, and proliferate.
The Background of Immune Repertoire Sequencing (Rep-seq)
DNA sequencing approaches have enabled the characterization of immune repertoires. A plethora of immune repertoire methods have been published and commercial solutions are also available. Basically, multiplex PCR can amplify the recombined V(D)J regions from either mRNA or DNA in the B- or T-cells. The V(D)J, and most importantly, the variable complementarity determining region CDR3 sequences, and their respective abundance can be resolved by high-throughput sequencing. Paired-end sequencing with additional PCR primers in the middle of the fragment permits full-length TCR repertoire sequencing with short read NGS technology to resolve the V/J pairing.
Rep-seq Service at Creative Biolabs
TCR sequencing (TCR-seq)
Our SuPrecision™ platform permits RNA as a template to amplify and sequence the complementary determining region (CDR) of whether α, β, γ or δ chain of TCR. Through this excellent platform, our clients can obtain the sequence data of the whole covered CDR3 regions of the interested TCR chain, which has involved most of the hypermutation information.
BCR sequencing (BCR-seq)
RNA from whole blood or B-cells is the required template for the amplification of CDR. Based on our advanced SuPrecision™ platform, both the heavy chain and the light chain CDR3 regions can be 100% covered and accurately sequenced, the constant regions from the two species are also captured and sequenced with enough length to identify the class information. In this way, our customers can finally obtain a comprehensive BCR repertoire involving both the hypermutation and isotype data from this sequencing service.
Rep-seq Workflow at Creative Biolabs
Bioinformatics Analysis at Creative Biolabs
Key Advantages of Rep-seq Service in Creative Biolabs Include but Are Not Limited to:
- Diverse species: human, mouse, rhesus monkey, alpaca, canine, rabbit, swine, feline, zebrafish, etc.
- High fidelity amplification method
- Specific capture primer
- Abundant sequencing platforms
- Accurate information analysis
A Wide Variety of Applications of Rep-seq Service Include but Are Not Limited to:
- Identification of antigen-specific TCR/BCR
- Minimal residual disease quantification
- Evaluation of the efficacy of immune checkpoint blockers
- Monitor immune diversity recovery after treatment
- Cancer immune therapy
Scientists from Creative Biolabs have accomplished over hundreds of sequencing projects and accumulated sufficient knowledge and understanding of the immune system. We are confident in providing the best service of Rep-seq at the single-cell level, and we also provide the most comprehensive germline genes and alleles identification service with the most accurate sequencing results for our global customers.
Please contact us for more information and a detailed quote.
- Lossius, A.; et al. High-throughput sequencing of immune repertoires in multiple sclerosis. Ann Clin Transl Neurol. 2016, 3(4):295-306.
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