Bibliografisk note

Funding Information:
We thank Drs Matthias Flotenmeyer and Lou Brillault from Center for Microscopy and Microanalysis (CMM), the University of Queensland, for facilitating cryo-EM work to be carried out and for scientific and technical assistance. We thank UQBR animal staff (Maya Patrick and Barb Arnts) and UQBR facility at the Australian Institute for Bioengineering and Nanotechnology (AIBN). We thank Research Computing Center (RCC), UQ, for partially providing computational resources. We thank Queensland Health Forensic and Scientific Services, Queensland, and the Kirby Institute University of New Souths Wales for providing SARS-CoV-2 virus isolates. We thank the National Biologics Facility, AIBN, for access to the Biacore 8K+ instrument for the SPR assay. We thank Prof. Andreas Suhrbier for providing Omicron BA.2 virus isolate. We thank Dr. Michael Landsberg for advice and helpful discussion. We thank Ian Mortimer (ITS Infrastructure Operations, UQ) for IT support. We thank the Max-Planck-Institut für molekulare Genetik (MPI-MG) for granting access to the TEM instrumentation of the microscopy and cryo-EM service group. N.M would like to thank Alpaca ‘Budda’ whose nanobody was isolated and Dzongsar Jamyang Khyentse Rinpoche for generous contribution. Funding. This work was supported by NHMRC MRFF Coronavirus Research Response grant APP1202445 to D.W. K.C. P.R.Y. and T.M. Cryo-EM equipment at CMM is supported by ARC The Linkage Infrastructure, Equipment and Facilities (LIEF) scheme. N.M. was supported by UQ Research Stimulus. D. S. was supported by the German Research Foundation (DFG) Emmy Noether Programme (SCHW1851/1-1) and by an EMBO Advanced grant (aALTF-1650). The Chilean platform for the generation and characterization of Camelid Nanobodies to A.R.F is funded by ANID-FONDECYT No. 1200427; the regional Council of the “Los Rios region” projects FICR19-20, FICR21-01; FICR20-49 to R.J; the Bio & Medical Technology Development Program of the National Research Foundation (NRF) of the Korean government (MSIT) (NRF-2020M3A9H5112395); the ANID-MPG MPG190011 and ANID-STINT CS2018-7952 grants; and the EU-LAC T010047. DB is funded by UKRI Biotechnology and Biological Sciences Research Council (UKRI - BBSRC, https://www.ukri.org/) Institute Strategic Program Grant (ISPG) to The Pirbright Institute (BBS/E/I/00007034, BBS/E/I/00007030 and BBS/E/I/00007039). N.M.: Experimental design; conceptualized; designed, generated, purified spike proteins, monoclonal antibodies, W25-Fc and control Fc; characterized spike proteins and nanobodies; generated W25/Omicron spike dataset including sample preparation, optimization and determination of optimal freezing conditions, image acquisition, performed single particle analysis (SPA); performed PRNTs; performed K18-huACE2 mouse challenge experiments; generated figures; drafted and edited manuscript. S.M.L.: Performed Wu spike Hexapro/W25 and Omicron spike Hexapro/W25 cryo-EM single particle and structure analysis. A.A.A.: Propagated live virus stock, performed PRNTs, performed K18-huACE2 mouse challenge experiments and data analysis. P.W.: Performed K18-huACE2 mouse challenge experiment. S.I.L.B.: Experimental design for the biodistribution studies, performed them with the help of J.T.J. and I.V.A. Y.S.L: Sample optimization for W25/Omicron spike cryo-EM, determination optimal freezing conditions, data acquisition, data analysis. N.T.: Performed cell-cell fusion experiments. B.L.: Assisted in cloning and purification of spike proteins and antibodies; assisted in K18-huACE2 mouse challenge experiments. G.V.N.: Identification of W25 cloning of W25 and performed thermostability and nebulization stability assays of W25. J.J.: Performed SPA analysis and focuses classification for W25/Omicron dataset. D.P.: Cloned, expressed, and purified published nanobodies. A.I.: Reformatted published nanobodies. J.D.S.: Generated live virus stock and generated VeroE6-TMPRSS6 cells for virus propagation. D.S.: Performed K18-huACE2 mouse challenge experiment. J.T.J.: Performed the biodistribution study with S.I.L.B and I.V.A. Y.C.: Performed optimization and purification of W25. J.B.: Assisted in Wu spike Hexapro/W25 cryo-EM single particle and structure analysis. I.V.A.: Performed the biodistribution study with S.I.L.B and J.T.J. J.H.: Performed optimization and purification of W25 for biodistribution assays. R.J.: Supervised vibrating mesh nebulizer stability assays. R.M.: Supervised vibrating mesh nebulizer stability assays. Z.M.C.: Performed optimization of W25. P.C.C.: supervised optimization of purification of W25 for cryo-EM and biodistribution. V.K.: Experimentally designed the biodistribution studies. T.M. and C.M.T.S.: Supervised Wu spike Hexapro/W25 cryo-EM single particle and structure analysis. A.A.K.: Obtained SARS-CoV-2 ancestral, Alpha, Beta isolates. Chief Investigator for BSL3 biocontainment facility at School of Chemistry and Molecular Biosciences. M.L.J.: Performed and analyzed SPR experiment. A.K.: Designed the biodistribution studies, supervised the work of J.T.J. M.M.H.: Designed the labeling and biodistribution studies, supervised the work of I.V.A. and S.I.L.B. K.A.J.: Supervised K18-huACE2 prophylactic mouse work. D. Sc.: Prepared and purified Wu Hexapro spike, prepared Wu spike Hexapro/W25 complex cryo-EM sample, performed and supervised Wu spike Hexapro/W25 cryo-EM and Omicron spike Hexapro/W25 single particle and structure analysis, generated figures, wrote initial manuscript draft. D.W.: Performed SPA of W25/Omicron dataset. The overall project was supervised, conceptualized, and edited by D.W. A.R.F. and D.S. D.W. K.C. and P.R.Y are listed as inventors of ‘Molecular Clamp’ patent, US 2020/0040042.

Funding Information:
We thank Drs Matthias Flotenmeyer and Lou Brillault from Center for Microscopy and Microanalysis (CMM), the University of Queensland, for facilitating cryo-EM work to be carried out and for scientific and technical assistance. We thank UQBR animal staff (Maya Patrick and Barb Arnts) and UQBR facility at the Australian Institute for Bioengineering and Nanotechnology (AIBN). We thank Research Computing Center (RCC), UQ, for partially providing computational resources. We thank Queensland Health Forensic and Scientific Services, Queensland, and the Kirby Institute University of New Souths Wales for providing SARS-CoV-2 virus isolates. We thank the National Biologics Facility, AIBN, for access to the Biacore 8K+ instrument for the SPR assay. We thank Prof. Andreas Suhrbier for providing Omicron BA.2 virus isolate. We thank Dr. Michael Landsberg for advice and helpful discussion. We thank Ian Mortimer (ITS Infrastructure Operations, UQ) for IT support. We thank the Max-Planck-Institut für molekulare Genetik (MPI-MG) for granting access to the TEM instrumentation of the microscopy and cryo-EM service group. N.M would like to thank Alpaca ‘Budda’ whose nanobody was isolated and Dzongsar Jamyang Khyentse Rinpoche for generous contribution. Funding. This work was supported by NHMRC MRFF Coronavirus Research Response grant APP1202445 to D.W., K.C., P.R.Y., and T.M. Cryo-EM equipment at CMM is supported by ARC The Linkage Infrastructure, Equipment and Facilities ( LIEF ) scheme. N.M. was supported by UQ Research Stimulus . D. S. was supported by the German Research Foundation (DFG) Emmy Noether Programme ( SCHW1851/1-1 ) and by an EMBO Advanced grant ( aALTF-1650 ). The Chilean platform for the generation and characterization of Camelid Nanobodies to A.R.F is funded by ANID-FONDECYT No. 1200427 ; the regional Council of the “ Los Rios region ” projects FICR19-20 , FICR21-01 ; FICR20-49 to R.J; the Bio & Medical Technology Development Program of the National Research Foundation (NRF) of the Korean government ( MSIT ) ( NRF-2020M3A9H5112395 ); the ANID-MPG MPG190011 and ANID-STINT CS2018-7952 grants; and the EU-LAC T010047 . DB is funded by UKRI Biotechnology and Biological Sciences Research Council (UKRI - BBSRC , https://www.ukri.org/ ) Institute Strategic Program Grant ( ISPG ) to The Pirbright Institute ( BBS/E/I/00007034 , BBS/E/I/00007030 and BBS/E/I/00007039 ).

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