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LitCovid-PubTator

LitCovid-PD-FMA-UBERON

LitCovid-PD-MONDO

LitCovid-PD-CLO

LitCovid-PD-CHEBI

LitCovid-PD-GO-BP

{"project":"LitCovid-PD-GO-BP","denotations":[{"id":"T4","span":{"begin":1072,"end":1077},"obj":"http://purl.obolibrary.org/obo/GO_0019835"}],"text":"Pseudovirus neutralization and inhibition assays\nSARS-CoV-2 pseudovirus was generated, as previously described, with some modifications.25,27,29 Briefly, 293T cells were cotransfected with a plasmid encoding Env-defective, luciferase-expressing HIV-1 genome (pNL4-3.luc.RE) and a plasmid encoding SARS-CoV-2 S protein using the calcium phosphate method. SARS-CoV and MERS-CoV pseudoviruses were packaged as controls. The transfected medium was changed into fresh Dulbecco’s Modified Eagle’s Medium (DMEM) 8 h later, and pseudovirus-containing supernatants were collected 72 h later for single-cycle infection in target cells. Pseudovirus neutralization assay was then performed by incubation of SARS-CoV-2, SARS-CoV, or MERS-CoV pseudovirus with serially diluted SARS-CoV RBD- or MERS-CoV RBD-immunized mouse sera for 1 h at 37 °C, followed by addition of the mixture into hACE2/293T (for SARS-CoV-2 pseudovirus and SARS-CoV pseudovirus) or hDPP4/293T (for MERS-CoV pseudovirus) target cells. Fresh medium was added 24 h later, and the cells were lysed 72 h later in cell lysis buffer (Promega, Madison, WI). The lysed cell supernatants were incubated with luciferase substrate (Promega) and detected for relative luciferase activity using the Infinite 200 PRO Luminator (Tecan). The 50% MERS pseudovirus neutralizing antibody titer (NT50) was calculated using the CalcuSyn computer program, as previously described.29,35\nInhibition of pseudovirus entry by SARS-CoV-2 RBD protein was carried out, as previously described, with some modifications.31 Briefly, SARS-CoV-2 RBD protein at serial dilutions was incubated with hACE2/293T target cells for 1 h at 37 °C. After removing medium containing the protein, the cells were infected with SARS-CoV-2 pseudovirus. SARS-CoV RBD and MERS-CoV RBD, as well as SARS-CoV pseudovirus and MERS-CoV pseudovirus, were used as controls. Fresh medium was added 24 h later, and the cells were lysed and analyzed, as described above. The 50% inhibitory concentration (IC50) of the RBD protein was calculated using the CalcuSyn computer program, as described above."}

LitCovid-sentences

{"project":"LitCovid-sentences","denotations":[{"id":"T167","span":{"begin":0,"end":48},"obj":"Sentence"},{"id":"T168","span":{"begin":49,"end":353},"obj":"Sentence"},{"id":"T169","span":{"begin":354,"end":416},"obj":"Sentence"},{"id":"T170","span":{"begin":417,"end":625},"obj":"Sentence"},{"id":"T171","span":{"begin":626,"end":992},"obj":"Sentence"},{"id":"T172","span":{"begin":993,"end":1108},"obj":"Sentence"},{"id":"T173","span":{"begin":1109,"end":1279},"obj":"Sentence"},{"id":"T174","span":{"begin":1280,"end":1421},"obj":"Sentence"},{"id":"T175","span":{"begin":1422,"end":1661},"obj":"Sentence"},{"id":"T176","span":{"begin":1662,"end":1760},"obj":"Sentence"},{"id":"T177","span":{"begin":1761,"end":1872},"obj":"Sentence"},{"id":"T178","span":{"begin":1873,"end":1966},"obj":"Sentence"},{"id":"T179","span":{"begin":1967,"end":2097},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Pseudovirus neutralization and inhibition assays\nSARS-CoV-2 pseudovirus was generated, as previously described, with some modifications.25,27,29 Briefly, 293T cells were cotransfected with a plasmid encoding Env-defective, luciferase-expressing HIV-1 genome (pNL4-3.luc.RE) and a plasmid encoding SARS-CoV-2 S protein using the calcium phosphate method. SARS-CoV and MERS-CoV pseudoviruses were packaged as controls. The transfected medium was changed into fresh Dulbecco’s Modified Eagle’s Medium (DMEM) 8 h later, and pseudovirus-containing supernatants were collected 72 h later for single-cycle infection in target cells. Pseudovirus neutralization assay was then performed by incubation of SARS-CoV-2, SARS-CoV, or MERS-CoV pseudovirus with serially diluted SARS-CoV RBD- or MERS-CoV RBD-immunized mouse sera for 1 h at 37 °C, followed by addition of the mixture into hACE2/293T (for SARS-CoV-2 pseudovirus and SARS-CoV pseudovirus) or hDPP4/293T (for MERS-CoV pseudovirus) target cells. Fresh medium was added 24 h later, and the cells were lysed 72 h later in cell lysis buffer (Promega, Madison, WI). The lysed cell supernatants were incubated with luciferase substrate (Promega) and detected for relative luciferase activity using the Infinite 200 PRO Luminator (Tecan). The 50% MERS pseudovirus neutralizing antibody titer (NT50) was calculated using the CalcuSyn computer program, as previously described.29,35\nInhibition of pseudovirus entry by SARS-CoV-2 RBD protein was carried out, as previously described, with some modifications.31 Briefly, SARS-CoV-2 RBD protein at serial dilutions was incubated with hACE2/293T target cells for 1 h at 37 °C. After removing medium containing the protein, the cells were infected with SARS-CoV-2 pseudovirus. SARS-CoV RBD and MERS-CoV RBD, as well as SARS-CoV pseudovirus and MERS-CoV pseudovirus, were used as controls. Fresh medium was added 24 h later, and the cells were lysed and analyzed, as described above. The 50% inhibitory concentration (IC50) of the RBD protein was calculated using the CalcuSyn computer program, as described above."}

2_test

{"project":"2_test","denotations":[{"id":"32203189-23978242-19774547","span":{"begin":136,"end":138},"obj":"23978242"},{"id":"32203189-19683779-19774548","span":{"begin":142,"end":144},"obj":"19683779"},{"id":"32203189-19683779-19774549","span":{"begin":1416,"end":1418},"obj":"19683779"},{"id":"32203189-16968952-19774550","span":{"begin":1419,"end":1421},"obj":"16968952"},{"id":"32203189-27750111-19774551","span":{"begin":1546,"end":1548},"obj":"27750111"}],"text":"Pseudovirus neutralization and inhibition assays\nSARS-CoV-2 pseudovirus was generated, as previously described, with some modifications.25,27,29 Briefly, 293T cells were cotransfected with a plasmid encoding Env-defective, luciferase-expressing HIV-1 genome (pNL4-3.luc.RE) and a plasmid encoding SARS-CoV-2 S protein using the calcium phosphate method. SARS-CoV and MERS-CoV pseudoviruses were packaged as controls. The transfected medium was changed into fresh Dulbecco’s Modified Eagle’s Medium (DMEM) 8 h later, and pseudovirus-containing supernatants were collected 72 h later for single-cycle infection in target cells. Pseudovirus neutralization assay was then performed by incubation of SARS-CoV-2, SARS-CoV, or MERS-CoV pseudovirus with serially diluted SARS-CoV RBD- or MERS-CoV RBD-immunized mouse sera for 1 h at 37 °C, followed by addition of the mixture into hACE2/293T (for SARS-CoV-2 pseudovirus and SARS-CoV pseudovirus) or hDPP4/293T (for MERS-CoV pseudovirus) target cells. Fresh medium was added 24 h later, and the cells were lysed 72 h later in cell lysis buffer (Promega, Madison, WI). The lysed cell supernatants were incubated with luciferase substrate (Promega) and detected for relative luciferase activity using the Infinite 200 PRO Luminator (Tecan). The 50% MERS pseudovirus neutralizing antibody titer (NT50) was calculated using the CalcuSyn computer program, as previously described.29,35\nInhibition of pseudovirus entry by SARS-CoV-2 RBD protein was carried out, as previously described, with some modifications.31 Briefly, SARS-CoV-2 RBD protein at serial dilutions was incubated with hACE2/293T target cells for 1 h at 37 °C. After removing medium containing the protein, the cells were infected with SARS-CoV-2 pseudovirus. SARS-CoV RBD and MERS-CoV RBD, as well as SARS-CoV pseudovirus and MERS-CoV pseudovirus, were used as controls. Fresh medium was added 24 h later, and the cells were lysed and analyzed, as described above. The 50% inhibitory concentration (IC50) of the RBD protein was calculated using the CalcuSyn computer program, as described above."}

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PMC:7091888 / 28769-30866 JSON TXT