Immune Repetoire Sequencing
Immune repertoire refers to the sum total of functionally diverse B and T-cells in the circulatory system at any given moment (Wang et al. 2011). The diversity of immune repertoire is of vital importance for health. Immune Repertoire Sequencing (IR-SEQ) is to amplify the complimentary determining region (CDR) of B-cell receptor (BCR) or T-cell receptor (TCR) using multiple-PCR or 5’ RACE methods, followed by high-throughput sequencing. It is used to study the diversity of the immune system and the associations between immune repertoire and diseases.
- Proprietary methods: we have designed specific primers for CDR3 V region and C region to amplify CDR3 region, and patents for TRA, IGH, and IGK/IGL have been applied successfully.
- High-fidelity amplification methods: multiple-PCR or 5’ RACE method is used to equivalently amplify different clones and the number of unique clones is as large as 105.
- Multiple sequencing platforms: Hiseq2000, Hiseq2500 and Roche 454 are available for different project requirements.
1. C. Wang et al. J. Immunol. 186, 65.20 (2011)
High-fidelity and low bias
Two-step library construction method and BGI proprietary primers for lower amplification bias and true T/B cell clone properties.
High consistency between two libraries from one sample (Fig. 1).
The following services are available for human T/B cells
BGI-developed software “IMonitor” demonstrates better performance in correcting PCR and sequencing errors, and providing more customized bioinformatic analysis.¹
Peripheral blood mononucleated cells (PBMCs) are isolated from peripheral blood, followed by DNA or RNA extraction. Multiple-PCR or 5’RACE method is used to capture CDR3 region (5’ RACE can also capture CDR1 and CDR2), and then the captured region is sequenced on high-throughput platforms (Hiseq2000, Hiseq2500, Miseq or Roche 454).
1. Zhang W, Du Y, et al. (2015). “IMonitor: A Robust Pipeline for TCR and BCR Repertoire Analysis.” Genetics.
The semi-quantitative results of V b and J b segment usage in blood of liver cancer patients and healthy adults.² (A) V segment usage in blood samples from 20 liver cancer patients. (B) J segment usage in blood samples from 20 liver cancer patients. (C) V segment usage in blood samples from 21 healthy adults. (D) J segment usage in blood samples from 21 healthy adults.
2. Yingxin Han, Xing Liu, et al. (2015). ” Identification of characteristic TRB V usage in HBV-associated HCC by using differential expression profiling analysis.” OncoImmunology.