Single Cell TCR Repatoir Analysis

Single Cell
TCR repertoireAnalysis

What is single cell TCR repertoire analysis?

What is single cell TCR repertoire analysis?

By integrating scRNA-seq and TCR repertoire analysis, gene expression information and TCRα/β pair sequences necessary for clone reconstitution can be simultaneously analyzed for individual T cells. Based on the characteristics and frequency of each clone, it is possible to narrow down clones that are promising for TCR gene therapy and other applications.

* For TCR repertoireanalysis, please also refer to the TCR repertoire analysis page. Here, TCR repertoireanalysis is performed at the single cell level.

ImmunoGeneTeqs

Single cell TCR repertoire analysis features

ImmunoGeneTeqs

Single cell TCR repertoire analysis features

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Single-cell RNA-seq is available with either whole transcriptome amplification (WTA) or targeted sequencing using BD's Immune Response Panel.

*Target sequencing can optionally analyze genes not included in the target panel.

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Capable of up to 20,000-30,000 cells/analysis, pairing efficiency of mouse and human bio-derived TCRα/β is around 20%-80% (depending on cell activation state).

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Simultaneous analysis with protein using BD's Abseq, BioLegend's TotalSeq, etc. is also available

TAS-Seq2 detects about 3 times more genes than BD's WTA kit

TAS-Seq2 has a higher T-cell receptor pairing rate than BD's WTA kit

sample

sample

TCR alpha total reads

TCR alpha total reads

TCR beta total reads

TCR beta total reads

TCR detected cells

TCR detected cells

TCR paired cell

TCR paired cell

TCR pairing rate

TCR pairing rate

BD WTA

BD WTA

12,955,106

12,955,106

9,457,677

9,457,677

1,076

1,076

613

613

57.0%

57.0%

TAS-Seq2

TAS-Seq2

12,569,275

12,569,275

12,890,758

12,890,758

1,156

1,156

1,005

1,005

86.9%

86.9%

Example of analysis of tumor-infiltrating CD8⁺T cells in a B16F10 subcutaneously implanted tumor mouse model upon anti-PD-L1 treatment (WTA)

Approx. 50,000 reads/cell

EXPRESSION PATTERNS OF REPRESENTATIVE CD8+ T CELL MARKER GENES

BD WTA

TAS-Seq2

TAS-Seq2

Clusters identified by TAS-Seq2

0: terminally exhausted, 1: effector, 2: proliferated, 3: exhausted progenitor-1, 4: exhausted progenitor-2, 5: naïve-like, 6: stem-like, 7: dying cell

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TAS-Seq2 can identify CD8+ T cell subsets that are difficult to identify with BD's WTA kit

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TAS-Seq2 can clearly detect genes involved in the regulation of CD8+ T cell function with a lower drop-out rate

scTCR Repertoire Analysis Workflow

scTCR Repertoire Analysis Workflow

Reverse transcription

PolyA addition to the 5' end


Self-hybridization and elongation


TCR Gene Amplification


TCR a/b library

Matters to be checked in advance

* For multiplex analysis, please refer to the FAQ page "About Multiplex Analysis".

Sample acceptance

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cDNA synthesized by our specified protocol (IGT will undertake cDNA amplification and subsequent steps)

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Pre-fractionated frozen T cell specimens (cDNA synthesis and beyond contracted by IGT)

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Unfractionated frozen cell specimens (IGT is entrusted with t cell preparation and beyond)

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Cryopreserved tissue specimens (IGT is entrusted with the preparation of cell suspensions and beyond)

* When cryopreserving cells and tissues, please use the preservation solution and freezing method specified by us.

How to send samples

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Please make sure that the sample is dry and leak-free, and send it to the shipping address below. (Available until 5:00 p.m. on weekdays)

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If shipping frozen tissue or cells, please include enough dry ice to maintain the frozen state and ship frozen.

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Please refer to "Sample Shipping Address" for shipping address.

* *Except for remote areas, please send by next morning (except Saturdays, Sundays, and national holidays).

Send samples to

〒277-0882
千葉県柏市柏の葉6丁目6番2号

三井リンクラボ柏の葉1 304号室

TEL:04-7192-8732

ImmunoGeneTeqs, Inc.

Lead Time

From sample receipt

2.5 months

Deliverables

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Work report

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Complete sequence data

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Mapping result files, etc.

The official report will be delivered on a hard drive at the time of delivery

A mapping analysis report will be included in the formal report

An example of the report can be seen in the example of the analysis of mouse tumor-infiltrating T cells

scTCR-seq (WTA)

Reference price

Number of samples

Number of samples

Number of cells/sample analyzed

Number of cells/sample analyzed

Price (incl. tax)

Price (incl. tax)

Single

Single

1

20,000

¥1,045,000 

¥1,045,000 

Multiplex

Multiplex

2

20,000

¥1,925,000 

¥1,925,000 

4

10,000 

10,000 

¥2,145,000 

¥2,145,000 

8

5,000

¥2,640,000 

¥2,640,000 

Analysis Examples

Analysis Examples

Analysis Examples

Example of analysis of mouse tumor-infiltrating T cells

Tumor-infiltrating T cells (14,100 cells) were classified into 10 clusters and visualized by dimensional compression; clones sharing the TCR were then defined and the properties of each clone were evaluated; data consisted of single cell RNA-seq data and single cell TCR repertoireanalysis data.

The actual report will also be in this style

Papers for Analysis and Use

Papers for Analysis and Use

Papers for Analysis and Use

Authors:Hiroyasu Aoki, Masahiro Kitabatake, Haruka Abe, Peng Xu, Mikiya Tsunoda, Shigeyuki Shichino, Atsushi Hara, Noriko Ouji-Sageshima, Chihiro Motozono, Toshihiro Ito, Kouji Matsushima, and Satoshi Ueha

Journal, 掲載日:Cell Reports, 2024年3月8日

Single Cell TCR Repertoire Analysis
Analysis Overview Hearing Form

Single Cell TCR Repertoire Analysis
Analysis Overview Hearing Form

If you would like to request an analysis, please fill out and submit the hearing form at

We will prepare a quotation based on the information you send us.