	US 12,012,670 B2
	High-throughput immune sequencing
George M. Church, Brookline, MA (US); Francois Vigneault, Medford, MA (US); Uri Laserson, Boston, MA (US); and Ido Bachelet, Brookline, MA (US)
Assigned to President and Fellows of Harvard College, Cambridge, MA (US)
Filed by President and Fellows of Harvard College, Cambridge, MA (US)
Filed on Jul. 17, 2019, as Appl. No. 16/513,945.
Application 16/513,945 is a continuation of application No. 13/878,400, granted, now 10,392,726, previously published as PCT/US2011/055801, filed on Oct. 11, 2011.
Claims priority of provisional application 61/391,337, filed on Oct. 8, 2010.
Prior Publication US 2020/0095704 A1, Mar. 26, 2020
Int. Cl. C12N 15/10 (2006.01); C07K 16/00 (2006.01); C07K 16/06 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6881 (2018.01); C12Q 1/6883 (2018.01); C40B 10/00 (2006.01); C40B 30/04 (2006.01); C40B 40/08 (2006.01); G16B 20/00 (2019.01); G16B 30/00 (2019.01); C12Q 1/6874 (2018.01)

CPC C40B 10/00 (2013.01) [C07K 16/00 (2013.01); C12N 15/10 (2013.01); C12Q 1/6806 (2013.01); C12Q 1/6881 (2013.01); C40B 30/04 (2013.01); C40B 40/08 (2013.01); G16B 20/00 (2019.02); G16B 30/00 (2019.02); C07K 16/065 (2013.01); C12Q 1/6874 (2013.01); C12Q 1/6883 (2013.01); C12Q 2535/00 (2013.01); C12Q 2535/122 (2013.01); C12Q 2600/112 (2013.01); C12Q 2600/158 (2013.01)]

14 Claims

1. A method comprising:

a) linking a polynucleotide sequence encoding a heavy chain variable region (VH) and a polynucleotide sequence encoding a light chain variable region (VL) from a single lymphocyte from a first biological sample from a subject, thereby forming first paired sequences; and performing high-throughput sequencing of the first paired sequences, thereby forming a first reference library of paired sequence reads,

b) linking a polynucleotide sequence encoding a heavy chain variable region (VH) and a polynucleotide sequence encoding a light chain variable region (VL) from a single lymphocyte from a second biological sample from the subject, thereby forming second paired sequences; and performing high-throughput sequencing of the second paired sequences, thereby forming a second reference library of paired sequence reads,

wherein the first biological sample is or has been obtained before an immune response of the subject and the second biological sample is or has been obtained during or after the immune response of the subject, and

c) comparing the paired sequence reads of the first reference library and the second reference library.