Creative Biolabs offers the unparalleled mass sequencing service to analyze the B cell receptors (BCR) repertoires. Our scientists can provide Magic™ single-cell BCR profiling by high throughput sequencing single B cell receptors. Above all, our method can keep the original linkage between the paired heavy and light chains intact. This technique increases the efficiency of identifying disease biomarkers and designing therapeutic antibodies.
Current BCR Single-cell Analysis Methods
B cell receptors help the immune system detect antigens and trigger antibody production. Every individual harbors a repertoire of BCR specific to a range of antigens to which the person has been previously exposed. Each BCR consists of a variable heavy (VH) and a variable light (VL) chain. Original paired VH:VL chains are important because they can be used as markers for antibodies. These are useful for applications such as tracking B cells, monitoring for minimal residual disease and diagnostics. The paired sequences have the clinical potential of rapidly improving vaccine design and identifying neutralizing antibodies. High-throughput next-generation sequencing of B cell receptor repertoires could provide important information, which is the key to understanding the adaptive immune responses and for diagnostic and therapeutic applications. Most high-throughput sequencing methods can identify large numbers of VH and VL, but they need large quantities of B cell DNA as starting material. Therefore, information about the endogenous pairing of heavy and light chains is lost after bulk lysis of B cell populations. Existing single cell-based sequencing approaches, such as scRT-PCR and Sanger sequencing, could avoid this problem, but they typically require complex instrumentation, and the process is complicated, expensive and time-consuming.
Fig.1 Overview of the high-throughput method for paired VH:VL single B cell receptor analysis (DeKosky 2013).
Single-cell BCR Profiling Service in Creative Biolabs
Creative Biolabs has designed a new single-cell sequencing approach that would retain the original VH:VL pairing with high throughput using low reagent and lowering cost. Overview of the high-throughput method for paired VH:VL single B cell receptor analysis is shown (Fig.1). Briefly, B-cell populations are sorted. Single cells are isolated and equilibrated with lysis buffer. Then mRNA is captured on magnetic beads, RT-PCR by emulsion VH:VL linkage PCR. Next-generation sequencing is performed on our Magic™ platform. Bioinformatic processing is used to analyze the paired VH:VL repertoire. A huge advantage for our approach is that the assay does not have to be done with live cells at the sites of the early steps. This approach for sequencing the paired VH:VL repertoire enables rapid interrogation of the immune response and can be applied to investigate, vaccine efficacy, immune system health and autoimmunity in clinical.
Key Advantages of Our Single-cell BCR profiling Service
The high-throughput identification of paired VH:VL also enables rapid generation of novel diagnostic, therapeutic or prophylactic antibodies. Scientists of Creative Biolabs have accumulated enough experience in high-throughput sequencing single-cell BCR with paired VH:VL. We are confident in providing clients with the best service at the most competitive cost.
Please contact us for more information and a detailed quote.