{"doi":"10.1101/2020.05.30.125484","title":"Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes","abstract":"The COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2 has led to accelerated efforts to develop therapeutics, diagnostics, and vaccines to mitigate this public health emergency. A key target of these efforts is the spike (S) protein, a large trimeric class I fusion protein that is metastable and difficult to produce recombinantly in large quantities. Here, we designed and expressed over 100 structure-guided spike variants based upon a previously determined cryo-EM structure of the prefusion SARS-CoV-2 spike. Biochemical, biophysical and structural characterization of these variants identified numerous individual substitutions that increased protein yields and stability. The best variant, HexaPro, has six beneficial proline substitutions leading to ~10-fold higher expression than its parental construct and is able to withstand heat stress, storage at room temperature, and multiple freeze-thaws. A 3.2 Å-resolution cryo-EM structure of HexaPro confirmed that it retains the prefusion spike conformation. High-yield production of a stabilized prefusion spike protein will accelerate the development of vaccines and serological diagnostics for SARS-CoV-2.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":118235,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":190,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9594,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":87308,"name":"Jory A. Goldsmith","orcid":"0000-0003-2917-0620","position":1,"is_corresponding":false},{"id":105917,"name":"Jeffrey M. Schaub","orcid":"0000-0002-1130-2675","position":2,"is_corresponding":false},{"id":105918,"name":"Andrea M. DiVenere","orcid":"0000-0001-9254-5347","position":3,"is_corresponding":false},{"id":262645,"name":"Hung‐Che Kuo","orcid":"0000-0002-8577-1706","position":4,"is_corresponding":false},{"id":105920,"name":"Kamyab Javanmardi","orcid":"0000-0002-6449-6709","position":5,"is_corresponding":false},{"id":105921,"name":"Kevin Le","orcid":"0000-0001-5098-7737","position":6,"is_corresponding":false},{"id":87305,"name":"Daniel Wrapp","orcid":"0000-0002-0538-9647","position":7,"is_corresponding":false},{"id":551130,"name":"Alison Gene-Wei Lee","orcid":null,"position":8,"is_corresponding":false},{"id":550084,"name":"Yutong Liu","orcid":"0000-0001-9386-3371","position":9,"is_corresponding":false},{"id":105924,"name":"Chia‐Wei Chou","orcid":"0000-0001-9618-338X","position":10,"is_corresponding":false},{"id":105925,"name":"Patrick O. Byrne","orcid":"0000-0002-6462-8951","position":11,"is_corresponding":false},{"id":105926,"name":"Christy K. Hjorth","orcid":"0000-0003-2324-8984","position":12,"is_corresponding":false},{"id":105927,"name":"Nicole V. Johnson","orcid":"0000-0003-4351-125X","position":13,"is_corresponding":false},{"id":105928,"name":"John Ludes-Meyers","orcid":null,"position":14,"is_corresponding":false},{"id":105929,"name":"Annalee W. Nguyen","orcid":"0000-0003-1268-7164","position":15,"is_corresponding":false},{"id":105930,"name":"Juyeon Park","orcid":"0000-0001-9549-4645","position":16,"is_corresponding":false},{"id":87306,"name":"Nianshuang Wang","orcid":"0000-0002-7569-011X","position":17,"is_corresponding":false},{"id":105931,"name":"Dzifa Amengor","orcid":"0000-0001-5705-3259","position":18,"is_corresponding":false},{"id":105934,"name":"Jennifer A. Maynard","orcid":"0000-0002-0363-8486","position":19,"is_corresponding":false},{"id":105935,"name":"Ilya J. Finkelstein","orcid":"0000-0002-9371-2431","position":20,"is_corresponding":false},{"id":87312,"name":"Jason S. McLellan","orcid":"0000-0003-3991-542X","position":21,"is_corresponding":false},{"id":87313,"name":"Ching‐Lin Hsieh","orcid":"0000-0002-3665-5717","position":0,"is_corresponding":true}],"reference_count":41,"raw_metadata":null,"created_at":"2026-07-18T23:13:58.531532Z","pmid":"32577660","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}