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

Abstract:

:Natively paired sequencing (NPS) of B cell receptors [variable heavy (VH) and light (VL)] and T cell receptors (TCRb and TCRa) is essential for the understanding of adaptive immunity in health and disease. Despite many recent technical advances, determining the VH:VL or TCRb:a 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 TCRb:a 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 TCRb:a.

journal_name

Sci Adv

journal_title

Science advances

authors

Tanno H,McDaniel JR,Stevens CA,Voss WN,Li J,Durrett R,Lee J,Gollihar J,Tanno Y,Delidakis G,Pothukuchy A,Ellefson JW,Goronzy JJ,Maynard JA,Ellington AD,Ippolito GC,Georgiou G

doi

10.1126/sciadv.aay9093

subject

Has Abstract

pub_date

2020-04-22 00:00:00

pages

eaay9093

issue

17

issn

2375-2548

pii

aay9093

journal_volume

6

pub_type

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