{"doi":"10.17504/protocols.io.n92ld8rnov5b/v1","title":"Small-scale Expression and Purification Protocol for His-SUMO Tagged Enterovirus D68 Strain STL 2014 12 3C Protease in E. coli v1","abstract":"This protocol describes the expression and purification of Enterovirus D68 (EV-D68) strain EV-D68 STL 2014 12 3C protease in E. coli Rosetta BL21(DE3)-RR cells. The protein is expressed with an N-terminal His-SUMO tag using an auto-induction system at 18°C. Purification involves cell lysis, initial nickel affinity chromatography (IMAC), SUMO tag removal using His-SENP1 protease, reverse IMAC to remove the cleaved tag, and a final size exclusion chromatography step using a Superdex 75 column. The protocol yields pure protein at approximately 20 mg/mL concentration, suitable for subsequent biochemical and structural studies. The entire process is optimized for small-scale expression (<6L) and takes approximately 5 days from transformation to final purified protein.","journal":null,"year":2024,"id":508460,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9559,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1059372,"name":"M. Fairhead","orcid":"0000-0001-5361-3933","position":1,"is_corresponding":false},{"id":557996,"name":"Eleanor Williams","orcid":"0000-0002-1331-9518","position":2,"is_corresponding":false},{"id":1339556,"name":"Korvus Wang","orcid":null,"position":0,"is_corresponding":true}],"reference_count":0,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:11:10.049781Z","pmid":null,"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":[]}