A facile technology for the high-throughput sequencing of the paired VH:VL and TCRβ:TCRα repertoires

Hidetaka Tanno, Jonathan R. McDaniel, Christopher A. Stevens, William N. Voss, Jie Li, Russell Durrett, Jiwon Lee, Jimmy Gollihar, Yuri Tanno, George Delidakis, Arti Pothukuchy, Jared W. Ellefson, Jörg J. Goronzy, Jennifer A. Maynard, Andrew D. Ellington, Gregory C. Ippolito, George Georgiou

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Natively paired sequencing (NPS) of B cell receptors [variable heavy (VH) and light (VL)] and T cell receptors (TCRβ and TCRα) is essential for the understanding of adaptive immunity in health and disease. Despite many recent technical advances, determining the VH:VL or TCRβ:α repertoire with high accuracy and throughput remains challenging. We discovered that the recently engineered xenopolymerase, RTX, is exceptionally resistant to cell lysate inhibition in single-cell emulsion droplets. We capitalized on the characteristics of this enzyme to develop a simple, rapid, and inexpensive in-droplet overlap extension reverse transcription polymerase chain reaction method for NPS not requiring microfluidics or other specialized equipment. Using this technique, we obtained high yields (5000 to >20,000 per sample) of paired VH:VL or TCRβ:α clonotypes at low cost. As a demonstration, we performed NPS on peripheral blood plasmablasts and T follicular helper cells following seasonal influenza vaccination and discovered high-affinity influenza-specific antibodies and TCRβ:α.

Original languageEnglish (US)
Article numberaay9093
JournalScience Advances
Volume6
Issue number17
DOIs
StatePublished - Apr 2020

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'A facile technology for the high-throughput sequencing of the paired VH:VL and TCRβ:TCRα repertoires'. Together they form a unique fingerprint.

Cite this