Eur J Immunol, 2013, 43(9):2507-2515

Pairing of T-cell receptor chains via emulsion PCR

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 mRNA molecules that are coexpressed in the same living cells. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

IBCH: 4417
Ссылка на статью в журнале: http://doi.wiley.com/10.1002/eji.201343453
Кол-во цитирований на 02.2024: 96
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