PMID- 32756607
OWN - NLM
STAT- MEDLINE
DCOM- 20201002
LR  - 20201002
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 15
IP  - 8
DP  - 2020
TI  - Cloning antibodies from single cells in pooled sequence libraries by selective 
      PCR.
PG  - e0236477
LID - 10.1371/journal.pone.0236477 [doi]
LID - e0236477
AB  - Antibodies function by binding to antigens. Antibodies must be cloned and 
      expressed to determine their binding characteristics, but current methods for 
      high-throughput antibody sequencing yield antibody DNA pooled from many cells and 
      do not readily permit cloning of antibodies from single B cells. We present a 
      strategy for retrieving and cloning antibody DNA from single cells within a 
      pooled library of cells. Our strategy, called selective PCR for antibody 
      retrieval (SPAR), takes advantage of the unique sequence barcodes attached to 
      individual cDNA molecules during sample preparation to enable specific 
      amplification by PCR of antibody heavy- and light-chain cDNA originating from a 
      single cell. We show through computational analysis that most human antibodies 
      sequenced using typical high-throughput methods can be retrieved using SPAR, and 
      experimentally demonstrate retrieval of full-length antibody variable region cDNA 
      from three cells within pools of ~5,000 cells. SPAR enables rapid low-cost 
      cloning and expression of native human antibodies from pooled single-cell 
      sequence libraries for functional characterization.
FAU - Horns, Felix
AU  - Horns F
AUID- ORCID: 0000-0001-5872-5061
AD  - Biophysics Graduate Program, Stanford University, Stanford, California, United 
      States of America.
FAU - Quake, Stephen R
AU  - Quake SR
AD  - Department of Bioengineering, Stanford University, Stanford, California, United 
      States of America.
AD  - Department of Applied Physics, Stanford University, Stanford, California, United 
      States of America.
AD  - Chan Zuckerberg Biohub, Stanford, California, United States of America.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200805
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Antibodies)
RN  - 0 (DNA, Complementary)
RN  - 0 (Immunoglobulin Heavy Chains)
RN  - 0 (Immunoglobulin Variable Region)
SB  - IM
MH  - Amino Acid Sequence/genetics
MH  - Antibodies/*genetics
MH  - Cell Surface Display Techniques
MH  - Cloning, Molecular
MH  - DNA, Complementary/genetics
MH  - Gene Library
MH  - Genetic Vectors/genetics
MH  - Humans
MH  - Immunoglobulin Heavy Chains/*genetics
MH  - Immunoglobulin Variable Region/*genetics
MH  - Polymerase Chain Reaction/*methods
MH  - Single-Cell Analysis
PMC - PMC7406036
COIS- F.H. and S.R.Q. are inventors on a patent applied for by Stanford University and 
      the Chan-Zuckerberg Biohub (CZB) (provisional patent application number 
      63/039,113) related to methods and compositions for selective PCR and cloning of 
      antibody sequences. S.R.Q. is employed as co-president of CZB. This does not 
      alter our adherence to PLOS ONE policies on sharing data and materials.
EDAT- 2020/08/07 06:00
MHDA- 2020/10/03 06:00
PMCR- 2020/08/05
CRDT- 2020/08/07 06:00
PHST- 2020/05/07 00:00 [received]
PHST- 2020/07/06 00:00 [accepted]
PHST- 2020/08/07 06:00 [entrez]
PHST- 2020/08/07 06:00 [pubmed]
PHST- 2020/10/03 06:00 [medline]
PHST- 2020/08/05 00:00 [pmc-release]
AID - PONE-D-20-13444 [pii]
AID - 10.1371/journal.pone.0236477 [doi]
PST - epublish
SO  - PLoS One. 2020 Aug 5;15(8):e0236477. doi: 10.1371/journal.pone.0236477. 
      eCollection 2020.