@article{70bb0b5676194c318640f4f954d16988,
title = "A potent and broad neutralization of SARS-CoV-2 variants of concern by DARPins",
abstract = "We report the engineering and selection of two synthetic proteins—FSR16m and FSR22—for the possible treatment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. FSR16m and FSR22 are trimeric proteins composed of DARPin SR16m or SR22 fused with a T4 foldon. Despite selection by a spike protein from a now historical SARS-CoV-2 strain, FSR16m and FSR22 exhibit broad-spectrum neutralization of SARS-CoV-2 strains, inhibiting authentic B.1.351, B.1.617.2 and BA.1.1 viruses, with respective IC50 values of 3.4, 2.2 and 7.4 ng ml−1 for FSR16m. Cryo-EM structures revealed that these DARPins recognize a region of the receptor-binding domain (residues 456, 475, 486, 487 and 489) overlapping a critical portion of the angiotensin-converting enzyme 2 (ACE2)-binding surface. K18-hACE2 transgenic mice inoculated with B.1.617.2 and receiving intranasally administered FSR16m showed less weight loss and 10–100-fold lower viral burden in upper and lower respiratory tracts. The strong and broad neutralization potency makes FSR16m and FSR22 promising candidates for the prevention and treatment of infection by SARS-CoV-2. [Figure not available: see fulltext.]",
author = "Vikas Chonira and Kwon, {Young D.} and Jason Gorman and Case, {James Brett} and Zhiqiang Ku and Rudo Simeon and Casner, {Ryan G.} and Harris, {Darcy R.} and Olia, {Adam S.} and Tyler Stephens and Lawrence Shapiro and Bender, {Michael F.} and Hannah Boyd and Teng, {I. Ting} and Yaroslav Tsybovsky and Florian Krammer and Ningyan Zhang and Diamond, {Michael S.} and Kwong, {Peter D.} and Zhiqiang An and Zhilei Chen",
note = "Funding Information: Prof. P. Bieniasz (The Rockefeller University) provided the 293T clone 22 cells for pseudoparticle neutralization assays. Prof. N. Landau (New York University) provided the plasmids for Δ19 spike protein of B.1.617.2 and C.37. Dr. A. Benjamin (Texas A&M University Health Science Center) assisted with Size Exclusion Chromatography experiments. We thank M. Lee for molecular dynamics analysis of RBD complexes and members of the Structural Biology Section, Vaccine Research Center, for comments and discussions. The following reagent was obtained through BEI Resources, NIAID, NIH: Spike Glycoprotein (Stabilized) from SARS-Related Coronavirus 2, Wuhan-Hu-1 with C-Terminal Histidine and Twin-Strep Tags, Recombinant from HEK293 Cells, NR-52724; Spike Glycoprotein Receptor Binding Domain (RBD) from SARS-Related Coronavirus 2, Wuhan-Hu-1 with C-Terminal Histidine Tag, Recombinant from HEK293 Cells, NR-52306. Funding for V.C., R.S. and Z.C. was provided by the National Institutes of Health Common Fund (grant DP2AI136600 to Z.C.). Funding for J.B.C. and M.S.D. was provided by the National Institute of Allergy and Infectious Diseases, National Institutes of Health and NIH (grants: R01 AI157155 to M.S.D.) J.B.C. was supported by a Helen Hay Whitney postdoctoral fellowship. Funding for Y.D.K., J.G., D.R.H., A.S.O., M.F.B., I.-T.T. and P.D.K. was provided by the Intramural Research Program of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health. R.G.C. and L.S. were supported in part by federal funds from LEIDOS under contract 18X144Q and Bill and Melinda Gates Foundation (INV-016167). Some of this work was performed at the Columbia University Cryo-EM Center at the Zuckerman Institute. T.S. and Y.T. were supported in part with federal funds from the Frederick National Laboratory for Cancer Research, NIH, under Contract HHSN261200800001. Some of the cryo-EM datasets were collected at the National CryoEM Facility (NCEF) of the National Cancer Institute, which was, in part, supported by the National Cancer Institute{\textquoteright}s National Cryo-EM Facility at the Frederick National Laboratory for Cancer Research under contract HSSN261200800001E. Funding for Z.K., H.B., N.Z. and Z.A was provided in part by a Welch Foundation endowment by grant AU-0042-20030616 and Cancer Prevention and Research Institute of Texas (CPRIT) by grants RP150551 and RP190561 (Z.A. and N.Z.). Reagent generation in the Krammer laboratory was partially funded by the NIAID Collaborative Influenza Vaccine Innovation Centers (CIVIC) under contract 75N93019C00051 and by the NIAID Centers of Excellence for Influenza Research and Response (CEIRR) under contracts 75N93021C00014 and 75N93021C00017. Funding Information: Prof. P. Bieniasz (The Rockefeller University) provided the 293T clone 22 cells for pseudoparticle neutralization assays. Prof. N. Landau (New York University) provided the plasmids for Δ19 spike protein of B.1.617.2 and C.37. Dr. A. Benjamin (Texas A&M University Health Science Center) assisted with Size Exclusion Chromatography experiments. We thank M. Lee for molecular dynamics analysis of RBD complexes and members of the Structural Biology Section, Vaccine Research Center, for comments and discussions. The following reagent was obtained through BEI Resources, NIAID, NIH: Spike Glycoprotein (Stabilized) from SARS-Related Coronavirus 2, Wuhan-Hu-1 with C-Terminal Histidine and Twin-Strep Tags, Recombinant from HEK293 Cells, NR-52724; Spike Glycoprotein Receptor Binding Domain (RBD) from SARS-Related Coronavirus 2, Wuhan-Hu-1 with C-Terminal Histidine Tag, Recombinant from HEK293 Cells, NR-52306. Funding for V.C., R.S. and Z.C. was provided by the National Institutes of Health Common Fund (grant DP2AI136600 to Z.C.). Funding for J.B.C. and M.S.D. was provided by the National Institute of Allergy and Infectious Diseases, National Institutes of Health and NIH (grants: R01 AI157155 to M.S.D.) J.B.C. was supported by a Helen Hay Whitney postdoctoral fellowship. Funding for Y.D.K., J.G., D.R.H., A.S.O., M.F.B., I.-T.T. and P.D.K. was provided by the Intramural Research Program of the Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health. R.G.C. and L.S. were supported in part by federal funds from LEIDOS under contract 18X144Q and Bill and Melinda Gates Foundation (INV-016167). Some of this work was performed at the Columbia University Cryo-EM Center at the Zuckerman Institute. T.S. and Y.T. were supported in part with federal funds from the Frederick National Laboratory for Cancer Research, NIH, under Contract HHSN261200800001. Some of the cryo-EM datasets were collected at the National CryoEM Facility (NCEF) of the National Cancer Institute, which was, in part, supported by the National Cancer Institute{\textquoteright}s National Cryo-EM Facility at the Frederick National Laboratory for Cancer Research under contract HSSN261200800001E. Funding for Z.K., H.B., N.Z. and Z.A was provided in part by a Welch Foundation endowment by grant AU-0042-20030616 and Cancer Prevention and Research Institute of Texas (CPRIT) by grants RP150551 and RP190561 (Z.A. and N.Z.). Reagent generation in the Krammer laboratory was partially funded by the NIAID Collaborative Influenza Vaccine Innovation Centers (CIVIC) under contract 75N93019C00051 and by the NIAID Centers of Excellence for Influenza Research and Response (CEIRR) under contracts 75N93021C00014 and 75N93021C00017. Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2023",
month = mar,
doi = "10.1038/s41589-022-01193-2",
language = "English",
volume = "19",
pages = "284--291",
journal = "Nature Chemical Biology",
issn = "1552-4450",
number = "3",
}