Emerging high-throughput sequencing methods for the analyses of complex structure of TCR and BCR repertoires give a powerful impulse to adaptive immunity studies. However, there are still essential technical obstacles for performing a truly quantitative analysis. Specifically, it remains challenging to obtain comprehensive information on the clonal composition of small lymphocyte populations, such as Ag-specific, functional, or tissue-resident cell subsets isolated by sorting, microdissection, or fine needle aspirates. In this study, we report a robust approach based on unique molecular identifiers that allows profiling Ag receptors for several hundred to thousand lymphocytes while preserving qualitative and quantitative information on clonal composition of the sample. We also describe several general features regarding the data analysis with unique molecular identifiers that are critical for accurate counting of starting molecules in high-throughput sequencing applications.

Central European Institute of Technology Masaryk University 601 77 Brno Czech Republic; and

Shemyakin Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences 117997 Moscow Russia;

Shemyakin Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences 117997 Moscow Russia; Central European Institute of Technology Masaryk University 601 77 Brno Czech Republic; and Pirogov Russian National Research Medical University 117997 Moscow Russia

Shemyakin Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences 117997 Moscow Russia; Faculty of Medicine Lomonosov Moscow State University 119192 Moscow Russia;

Vavilov Institute of General Genetics Russian Academy of Sciences 119991 Moscow Russia;

Comment In

Oncotarget. 2015 Nov 24;6(37):39393-4. doi: 10.18632/oncotarget.6349. PubMed

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