Overview of methodologies for T-cell receptor repertoire analysis

BMC Biotechnology

Table 1 Exemplary companies providing immune repertoire products and services

Company	Service/Kit	Starting material	Library preparation approach	Chains	CDR regions	Organism	Sequencing platform and length (bp)
BGI (Shenzhen, China)	Service	gDNA RNA	Mplex-PCR: primers V- C genes	TCRαTCRβ BCRH, BCRL	CDR3	Human	Illumina: HiSeq2000/2500 (100 × 2 bp) MiSeq (150/300 × 2 bp) Roche454
BGI (Shenzhen, China)	Service	RNA	5’RACE	TCRα, TCRβ	CDR1 CDR2 CDR3	Human
Adaptive Biotechnologies-ImmunoSeq (Seattle, USA)	Service Kit	gDNA cDNA	Mplex-PCR: primers V-J genes	TCRα, TCRβ, TCRδ, TCRγ, BCRH, BCRL, BCRK	CDR3	Human Mouse	Illumina: HiSeq, MiSeq
iRepertoire, Inc. (Huntsville, USA)	Service Kit	gDNA	Mplex-PCR: primers V-J genes	TCRβ	CDR3	Human	Illumina: HiSeq, MiSeq (100/150 × 2 bp)
iRepertoire, Inc. (Huntsville, USA)	Service Kit	RNA	Mplex-PCR: primers V-C genes	TCRα, TCRβ, TCRδ, TCRγ, BCRH, BCRL	CDR2 CDR3	Human Mouse	Illumina: HiSeq, MiSeq (100/150/250 × 2 bp) Roche454 (500 bp)
Clonotech Takara Bio USA, Inc. (Mountain View, USA)	Kit	RNA	SMART technology (5’RACE)	TCRα, TCRβ	CDR1 CDR2 CDR3	Human Mouse	Illumina: HiSeq, MiSeq Used by company for validation: MiSeq (300 × 2 bp)

Many different features are available; these can be combined in more than one way. Choice of primers, sequencing platform and depth may vary depending on starting material and desired outcome. Adaptive Biotechnologies only uses cDNA for limited applications. The company applies primer concentration controls for amplification bias correction and different options for sequencing depth are available (survey, deep, ultra-deep, max depth). iRepertoire may offer sequencing of the CDR2 region, depending on the chosen sequencing length

ISSN: 1472-6750