Directed evolution of super-secreted variants from phage-displayed human Interleukin-2.
Amino Acid Substitution
Animals
Antineoplastic Agents
/ pharmacology
Cell Line, Tumor
Cell Proliferation
/ drug effects
Cell Surface Display Techniques
Cell Survival
/ drug effects
Evolution, Molecular
Humans
Interleukin-2
/ genetics
Mice
Mice, Inbred C57BL
Protein Engineering
Receptors, Fc
/ drug effects
Recombinant Fusion Proteins
/ pharmacology
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
28 01 2019
28 01 2019
Historique:
received:
05
06
2018
accepted:
30
11
2018
entrez:
30
1
2019
pubmed:
30
1
2019
medline:
30
7
2020
Statut:
epublish
Résumé
Selection from a phage display library derived from human Interleukin-2 (IL-2) yielded mutated variants with greatly enhanced display levels of the functional cytokine on filamentous phages. Introduction of a single amino acid replacement selected that way (K35E) increased the secretion levels of IL-2-containing fusion proteins from human transfected host cells up to 20-fold. Super-secreted (K35E) IL-2/Fc is biologically active in vitro and in vivo, has anti-tumor activity and exhibits a remarkable reduction in its aggregation propensity- the major manufacturability issue limiting IL-2 usefulness up to now. Improvement of secretion was also shown for a panel of IL-2-engineered variants with altered receptor binding properties, including a selective agonist and a super agonist that kept their unique properties. Our findings will improve developability of the growing family of IL-2-derived immunotherapeutic agents and could have a broader impact on the engineering of structurally related four-alpha-helix bundle cytokines.
Identifiants
pubmed: 30692603
doi: 10.1038/s41598-018-37280-5
pii: 10.1038/s41598-018-37280-5
pmc: PMC6349883
doi:
Substances chimiques
Antineoplastic Agents
0
IL2 protein, human
0
Interleukin-2
0
Receptors, Fc
0
Recombinant Fusion Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
800Références
Chem Rev. 1997 Apr 1;97(2):391-410
pubmed: 11848876
Protein Eng. 2003 Dec;16(12):1081-7
pubmed: 14983090
Nat Rev Immunol. 2004 Sep;4(9):665-74
pubmed: 15343366
J Immunol Methods. 2004 Oct;293(1-2):71-83
pubmed: 15541278
Science. 2005 Nov 18;310(5751):1159-63
pubmed: 16293754
Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2788-93
pubmed: 16477002
Science. 2006 Mar 31;311(5769):1924-7
pubmed: 16484453
J Mol Biol. 2007 Feb 16;366(2):449-60
pubmed: 17174328
J Am Chem Soc. 2007 Aug 22;129(33):10110-2
pubmed: 17665911
Cancer. 2008 Jul 15;113(2):293-301
pubmed: 18457330
J Biol Chem. 1991 Oct 25;266(30):20363-8
pubmed: 1939091
J Immunother. 2009 Nov-Dec;32(9):887-94
pubmed: 19816193
Biophys J. 2010 May 19;98(10):2309-16
pubmed: 20483340
Protein Eng Des Sel. 2011 Mar;24(3):283-90
pubmed: 21109548
Clin Cancer Res. 2011 Jun 1;17(11):3673-85
pubmed: 21531812
N Engl J Med. 2011 Dec 1;365(22):2055-66
pubmed: 22129252
Nature. 2012 Mar 25;484(7395):529-33
pubmed: 22446627
Immunobiology. 2013 Jan;218(1):105-13
pubmed: 22459271
Proc Natl Acad Sci U S A. 2012 Jul 3;109(27):10879-84
pubmed: 22745168
Biophys J. 2012 Apr 18;102(8):1907-15
pubmed: 22768947
Protein J. 2012 Oct;31(7):529-43
pubmed: 22791129
Biochem Soc Trans. 2012 Oct;40(5):1032-7
pubmed: 22988860
Cell Rep. 2012 Nov 29;2(5):1425-37
pubmed: 23168257
Curr Top Med Chem. 2012;12(22):2470-8
pubmed: 23339301
J Immunol. 2013 Jun 15;190(12):6230-8
pubmed: 23677467
BMC Biotechnol. 2013 Jun 26;13:52
pubmed: 23802841
Mol Syst Biol. 2013;9:689
pubmed: 24045637
MAbs. 2014 Jan 1;6(1):273-85
pubmed: 24253188
Sci Rep. 2013 Nov 26;3:3333
pubmed: 24276756
J Clin Invest. 2014 Jan;124(1):99-110
pubmed: 24292706
MAbs. 2014 May-Jun;6(3):637-48
pubmed: 24589624
Lancet Diabetes Endocrinol. 2013 Dec;1(4):295-305
pubmed: 24622415
J Pharm Sci. 2015 Feb;104(2):307-26
pubmed: 25492409
J Mol Recognit. 2015 Apr;28(4):261-8
pubmed: 25683569
Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3320-5
pubmed: 25733854
Cancer Cell. 2015 Apr 13;27(4):489-501
pubmed: 25873172
Immunity. 2015 May 19;42(5):826-38
pubmed: 25992859
J Chem Theory Comput. 2008 Mar;4(3):435-47
pubmed: 26620784
Clin Cancer Res. 2016 Feb 1;22(3):680-90
pubmed: 26832745
Oncoimmunology. 2017 Jan 11;6(3):e1277306
pubmed: 28405498
Nat Commun. 2017 May 12;8:15373
pubmed: 28497796
Methods Mol Biol. 2018;1701:535-560
pubmed: 29116526
J Immunol. 2018 May 15;200(10):3475-3484
pubmed: 29618524
J Immunol. 1978 Jun;120(6):2027-32
pubmed: 307029
Biochem J. 1987 Jul 1;245(1):85-91
pubmed: 3499142
Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92
pubmed: 3881765
Nucleic Acids Res. 1983 Jul 11;11(13):4307-23
pubmed: 6306584
Immunol Rev. 1980;51:337-57
pubmed: 7000676
J Gen Microbiol. 1993 Oct;139(10):2465-73
pubmed: 8254317
Arch Biochem Biophys. 1997 Mar 1;339(1):79-84
pubmed: 9056236
Biotherapy. 1997;10(1):15-24
pubmed: 9261546
Cytokine. 1997 Oct;9(10):734-9
pubmed: 9344504
Immunotechnology. 1997 Oct;3(3):185-93
pubmed: 9358271