A high throughput bispecific antibody discovery pipeline.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
07 04 2023
07 04 2023
Historique:
received:
20
02
2023
accepted:
22
03
2023
medline:
11
4
2023
entrez:
7
4
2023
pubmed:
8
4
2023
Statut:
epublish
Résumé
Bispecific antibodies (BsAbs) represent an emerging class of immunotherapy, but inefficiency in the current discovery has limited their broad clinical availability. Here we report a high throughput, agnostic, single-cell-based functional screening pipeline, comprising molecular and cell engineering for efficient generation of BsAb library cells, followed by functional interrogation at the single-cell level to identify and sort positive clones and downstream sequence identification and functionality characterization. Using a CD19xCD3 bispecific T cell engager (BiTE) as a model, we demonstrate that our single-cell platform possesses a high throughput screening efficiency of up to one and a half million variant library cells per run and can isolate rare functional clones at a low abundance of 0.008%. Using a complex CD19xCD3 BiTE-expressing cell library with approximately 22,300 unique variants comprising combinatorially varied scFvs, connecting linkers and VL/VH orientations, we have identified 98 unique clones, including extremely rare ones (~ 0.001% abundance). We also discovered BiTEs that exhibit novel properties and insights to design variable preferences for functionality. We expect our single-cell platform to not only increase the discovery efficiency of new immunotherapeutics, but also enable identifying generalizable design principles based on an in-depth understanding of the inter-relationships between sequence, structure, and function.
Identifiants
pubmed: 37029216
doi: 10.1038/s42003-023-04746-w
pii: 10.1038/s42003-023-04746-w
pmc: PMC10082157
doi:
Substances chimiques
Antibodies, Bispecific
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
380Informations de copyright
© 2023. The Author(s).
Références
MAbs. 2019 Aug/Sep;11(6):1025-1035
pubmed: 31185801
MAbs. 2023 Jan-Dec;15(1):2153410
pubmed: 36472472
Int J Cancer. 2002 Aug 20;100(6):690-7
pubmed: 12209608
Cancer Res. 2008 Jan 1;68(1):143-51
pubmed: 18172306
Front Immunol. 2017 Jul 10;8:793
pubmed: 28740495
Blood. 2011 Apr 28;117(17):4542-51
pubmed: 21300981
Autoimmun Rev. 2019 Feb;18(2):123-128
pubmed: 30572136
J Immunol. 2008 May 15;180(10):7028-38
pubmed: 18453625
Cell Rep. 2018 Feb 20;22(8):2206-2215
pubmed: 29466744
Lab Chip. 2009 Sep 21;9(18):2628-31
pubmed: 19704976
Protein Eng Des Sel. 2013 May;26(5):359-67
pubmed: 23468569
Lab Chip. 2012 Jun 21;12(12):2146-55
pubmed: 22318506
Hybridoma. 1984 Winter;3(4):305-20
pubmed: 6441771
Cancer Res. 2009 Jun 15;69(12):4941-4
pubmed: 19509221
Expert Opin Biol Ther. 2015;15(8):1093-9
pubmed: 25971805
Sci Rep. 2020 Aug 24;10(1):14103
pubmed: 32839506
Ann N Y Acad Sci. 1991 Dec 30;636:288-94
pubmed: 1793216
Lab Chip. 2018 Dec 4;18(24):3733-3749
pubmed: 30397689
Curr Opin Chem Biol. 2013 Jun;17(3):400-5
pubmed: 23683347
RSC Adv. 2020 Jul 20;10(45):27006-27013
pubmed: 35515810
Sci Adv. 2021 Jun 11;7(24):
pubmed: 34117053
MAbs. 2011 Nov-Dec;3(6):546-57
pubmed: 22123055
Cancer Immunol Immunother. 1997 Nov-Dec;45(3-4):190-2
pubmed: 9435871
Mol Cancer Ther. 2018 Apr;17(4):776-785
pubmed: 29339550
MAbs. 2016 Jul;8(5):892-904
pubmed: 27049350
Drug Discov Today. 2015 May;20(5):588-94
pubmed: 25757598
Proteins. 2018 May;86(5):495-500
pubmed: 29490423
Nat Protoc. 2013 May;8(5):870-91
pubmed: 23558786
Biochem Biophys Res Commun. 2004 May 28;318(2):507-13
pubmed: 15120630
MAbs. 2010 Mar-Apr;2(2):129-36
pubmed: 20190561
Cancer Cell. 2017 Mar 13;31(3):383-395
pubmed: 28262555
Nat Med. 2019 Sep;25(9):1408-1414
pubmed: 31477906
Sci Rep. 2017 Sep 11;7(1):11217
pubmed: 28894181
Biochemistry. 2019 Dec 3;58(48):4869-4881
pubmed: 31702909
Mol Immunol. 2007 Mar;44(8):1935-43
pubmed: 17083975
Nat Chem Biol. 2016 Feb;12(2):76-81
pubmed: 26641932
Ther Adv Vaccines Immunother. 2018 Feb;6(1):3-17
pubmed: 29998217
J Immunol. 2019 Aug 1;203(3):585-592
pubmed: 31332079
Cancer Immunol Immunother. 2010 Aug;59(8):1197-209
pubmed: 20309546
Nat Rev Cancer. 2011 Dec 01;12(1):51-8
pubmed: 22129804
Annu Rev Med. 2019 Jan 27;70:437-450
pubmed: 30379598
JCI Insight. 2018 Feb 22;3(4):
pubmed: 29467338
Methods. 2019 Feb 1;154:102-117
pubmed: 30395966
J Pharmacol Exp Ther. 2015 Nov;355(2):199-205
pubmed: 26341624
J Biol Chem. 2010 Jul 2;285(27):20850-9
pubmed: 20444694
Drug Discov Today. 2005 Sep 15;10(18):1237-44
pubmed: 16213416
Adv Drug Deliv Rev. 2013 Oct;65(10):1357-69
pubmed: 23026637
Int Immunopharmacol. 2012 Dec;14(4):770-8
pubmed: 23092552
Proc Natl Acad Sci U S A. 2018 Jun 12;115(24):E5467-E5476
pubmed: 29844189
Sci Rep. 2017 Jun 6;7(1):2862
pubmed: 28588218
Nat Commun. 2020 Oct 2;11(1):4974
pubmed: 33009381
Proc Natl Acad Sci U S A. 2016 Jan 26;113(4):E450-8
pubmed: 26759368
Antibodies (Basel). 2016 Mar 04;5(1):
pubmed: 31557987
MAbs. 2012 Nov-Dec;4(6):653-63
pubmed: 22925968
Blood Adv. 2018 Jun 12;2(11):1250-1258
pubmed: 29858209
PLoS One. 2019 Jul 25;14(7):e0219842
pubmed: 31344144
Comput Struct Biotechnol J. 2020 May 14;18:1221-1227
pubmed: 32542108
N Biotechnol. 2017 Oct 25;39(Pt B):167-173
pubmed: 28137467
PLoS One. 2013;8(2):e57479
pubmed: 23468998
Sci Signal. 2011 Jun 07;4(176):ra39
pubmed: 21653229
J Clin Oncol. 2016 Apr 1;34(10):1104-11
pubmed: 26884582
BioDrugs. 2020 Apr;34(2):111-119
pubmed: 31916225
Mol Immunol. 2011 Dec;49(3):474-82
pubmed: 22014687
Protein Eng. 2001 Aug;14(8):529-32
pubmed: 11579220
Protein Eng Des Sel. 2009 Mar;22(3):135-47
pubmed: 19188138
Immunol Rev. 2010 May;235(1):55-72
pubmed: 20536555
MAbs. 2015;7(3):584-604
pubmed: 25875246
Nat Biotechnol. 2020 Jun;38(6):715-721
pubmed: 32231335
Nat Rev Drug Discov. 2019 Aug;18(8):585-608
pubmed: 31175342
Immunotherapy. 2016 Jul;8(8):889-906
pubmed: 27161438