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- Title
Pairing of T-cell receptor chains via emulsion PCR.
- Authors
Turchaninova, Maria A.; Britanova, Olga V.; Bolotin, Dmitriy A.; Shugay, Mikhail; Putintseva, Ekaterina V.; Staroverov, Dmitriy B.; Sharonov, George; Shcherbo, Dmitriy; Zvyagin, Ivan V.; Mamedov, Ilgar Z.; Linnemann, Carsten; Schumacher, Ton N.; Chudakov, Dmitriy M.
- Abstract
Our ability to analyze adaptive immunity and engineer its activity has long been constrained by our limited ability to identify native pairs of heavy-light antibody chains and alpha-beta T-cell receptor ( TCR) chains - both of which comprise coupled 'halves of a key', collectively capable of recognizing specific antigens. Here, we report a cell-based emulsion RT-PCR approach that allows the selective fusion of the native pairs of amplified TCR alpha and beta chain genes for complex samples. A new type of PCR suppression technique was developed that makes it possible to amplify the fused library with minimal noise for subsequent analysis by high-throughput paired-end Illumina sequencing. With this technique, single analysis of a complex blood sample allows identification of multiple native TCR chain pairs. This approach may be extended to identify native antibody chain pairs and, more generally, pairs of m RNA molecules that are coexpressed in the same living cells.
- Publication
European Journal of Immunology, 2013, Vol 43, Issue 9, p2507
- ISSN
0014-2980
- Publication type
Article
- DOI
10.1002/eji.201343453