TCR and BCR repertoires sequencing: TCR Rep-Seq and BCR Rep-Seq
Our Service NGS-Repertoire7 provides sequencing and analysis of TCR and BCR repertoires.
NGS sequencing allows to analyze T Cell Receptor and B Cell Receptor repertoires in an unprecedented way. Doing amplicons from blood samples tens of thousands of clonal sequences can be obtained. This information can be analyzed in different ways depending on the goal of the experiments.
- Diversity index. A score can be obtained representing the degree of clonal diversity of the sample
- Comparing points in a time frame. This is very useful when analyzing changes in the T-cell repertoire in a process. This process could be an Infection, an autoimmune disease or an analysis after vaccination.
Tumor immunology and immunotherapy could be a target of TCR repertoire sequencing. The results sequencing TILs (Tumor Infiltrating Lymphocytes) repertoires can be compared with results sequencing blood samples.
DNA-based or RNA-based approaches
We offer two different approaches: DNA-based and RNA-based.
This is a good option if it is possible to obtain good quality RNA from your sample, e. g. fresh blood. Using this method you can obtain full-length immune receptor RNA sequence.
This is a robust approach that can be used in almost all kind of samples. We have developed our own multiplex PCR approach that allows the accurate sequencing of the CDR3 segment and the identification of the V and J alleles.
Follow-up across multiple time points
There is not "normal" or "healthy" repertoire to be used as reference an, thus, to follow-up a patient TCR/BCR repertoire across multiple time points it is a very informative approach. We have special graphical representations and methods to compare repertoires of the same patient at different time points.
TCR and BCR repertoires in different scenarios
There are many different scenarios in which you can want to know the immune repertoires. We have selected some of them.
TCR repertoires in Formalin-Fixed Paraffin-Embedded (FFPE) samples
It is very important to know which is the actual T cell repertoire in tumor samples. The sample is many times an FFPE sample. In this case, given that the RNA obtained from this kind of samples is not a good quality RNA, we recommend DNA-based approaches. In fact, we have developed our own DNA-based multiplex PCR approach especially designed for being used with FFPE samples.
Selection of special areas of the slide using Laser Microdissection
In this scenario we also can offer the specific analysis of special areas of the slide that we can obtain by means of Laser Microdissection. This means that you can analyze TCR repertoires from different areas of the tumor and of the stromal regions. We could even separate different subpopulations of T cells based on biomarkers. For example, we could analyze repertoires of CD4+ cells separated from CD8+ cells, or other subsets defined with other specific differential biomarkers.
TCR and BCR whole blood repertoire
This is a very frequent scenario in which we can analyze all the cells present in the blood sample. In this case, both approaches, RNA-based and DNA-based, can be used.
TCR and BCR repertoires in different Peripheral Blood Mononuclear Cells (PBMC) subpopulations
Sometimes it could be very informative to discriminate between different lymphocyte subpopulations and get repertorire information restricted to different subsets of lymphocytes. We can offer this service.