Gas chromatography-mass spectrometry method for determination of β-propiolactone in human inactivated rabies vaccine and its hydrolysis analysis

Lei, Shuo; Gao, Xun; Sun, Yang; Yu, Xiangyong; Zhao, Longshan

doi:10.1016/j.jpha.2018.06.003

Cited by 19 publications

(8 citation statements)

References 10 publications

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“…To validate the efficiency, a stock solution was divided into an untreated fraction (control) and a fraction, which was treated with 0.1% ß-propiolactone. After 16 h of incubation at 4 °C, samples were transferred to 37 °C for 2 h to hydrolyze all residual ß-propiolactone to prevent cytotoxicity to mammalian cells 49 . After hydrolysis, samples were inoculated onto 293T-hsACE2 cells, and incubated at 37 °C with 5% CO 2 in a humidified incubator.…”

Section: Methodsmentioning

confidence: 99%

Characterization of binding interactions of SARS-CoV-2 spike protein and DNA-peptide nanostructures

Kruse

Altattan

Laux

et al. 2022

Sci Rep

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Binding interactions of the spike proteins of the severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) to a peptide fragment derived from the human angiotensin converting enzyme 2 (hACE2) receptor are investigated. The peptide is employed as capture moiety in enzyme linked immunosorbent assays (ELISA) and quantitative binding interaction measurements that are based on fluorescence proximity sensing (switchSENSE). In both techniques, the peptide is presented on an oligovalent DNA nanostructure, in order to assess the impact of mono- versus trivalent binding modes. As the analyte, the spike protein and several of its subunits are tested as well as inactivated SARS-CoV-2 and pseudo viruses. While binding of the peptide to the full-length spike protein can be observed, the subunits RBD and S1 do not exhibit binding in the employed concentrations. Variations of the amino acid sequence of the recombinant full-length spike proteins furthermore influence binding behavior. The peptide was coupled to DNA nanostructures that form a geometric complement to the trimeric structure of the spike protein binding sites. An increase in binding strength for trimeric peptide presentation compared to single peptide presentation could be generally observed in ELISA and was quantified in switchSENSE measurements. Binding to inactivated wild type viruses could be shown as well as qualitatively different binding behavior of the Alpha and Beta variants compared to the wild type virus strain in pseudo virus models.

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Section: Methodsmentioning

confidence: 99%

Characterization of binding interactions of SARS-CoV-2 spike protein and DNA-peptide nanostructures

Kruse

Altattan

Laux

et al. 2022

Sci Rep

View full text Add to dashboard Cite

show abstract

“…After incubation at 4C for 16 hours, samples were transferred to 37C for two (2) hours to hydrolyze all residual beta-propiolactone. This step ensures complete hydrolysis of beta-propiolactone to prevent cytotoxicity to mammalian cells [7]. After hydrolysis, samples were inoculated onto Vero E6 cells, and incubated at 37C, 5% CO2 incubator and monitored daily for CPE.…”

Section: Methodsmentioning

confidence: 99%

Propagation, inactivation, and safety testing of SARS-CoV-2

Jureka

Silvas

Basler

2020

Preprint

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In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-9 CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has 10 been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 11 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease 12 that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly 13 described and effective procedures that enable the propagation and quantification of infectious 14 virus. Because work with the virus is recommended to be performed at biosafety level 3, validated 15 methods to effectively inactivate the virus to enable safe study of RNA, DNA and protein from 16 infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell 17 lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline 18 cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate 19 effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat. 20 cells to characterize viral genome sequences, monitor viral gene expression and genome replication, 43 and to characterize host responses to infection. Removal of intact, virus-infected cells is critical for 44 studies involving microscopy aimed at understanding the SARS-CoV-2:host interplay at the cellular 45 level, and also for high-throughput analysis of the progression of viral replication in response to 46 antivirals. Whole inactivated virus and viral proteins are needed for the development of inactivated 47 whole-virus vaccine preparations and also as a source of antigen for immunoassays. 48To help address these needs and to facilitate SARS-CoV-2 research efforts, we describe here 49 methods for the propagation of SARS-CoV-2 in multiple cell lines. We have also determined a more 50 efficient method for quantifying virus by plaque assay and have developed a SARS-CoV-2-specific 51 focus forming assay which can enhance throughput of assays requiring quantification of viral titers. 52Additionally, we describe validation if methods for the inactivation of SARS-CoV-2 through the use 53 of TRIzol, 10% neutral buffered formalin, beta-propiolactone, and heat. Taken together, the data 54 presented here will serve to provide researchers with a helpful basis of information to aid in their 55 work on SARS-CoV-2. 56 57 2. Materials and Methods 58 2.1 Cells and Virus 59 60 Vero E6 (ATCC# CRL-1586), Calu-3 (ATCC# HTB-55), Caco-2 (ATCC# HTB-37), Huh7, A549 61 (ATCC# CCL-185), and 293T cells were maintained in DMEM (Corning) supplemented with 10% heat 62 inactivated fetal bovine serum (FBS; GIBCO). Cells were kept in a 37°C, 5% CO2 incubator without 63 antibiotics or antimycotics. SARS-CoV-2, strain USA_WA1/2020, was obtained from the World 64 Reference Collection for Emerging Viruses and Arboviruses at the University of Texas Medical 65 Branch-Galveston. 6...

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“…After incubation at 4 • C for 16 h, samples were transferred to 37 • C for two (2) hours to hydrolyze all residual beta-propiolactone. This step ensures complete hydrolysis of beta-propiolactone to prevent cytotoxicity to mammalian cells [7]. After hydrolysis, samples were inoculated onto Vero E6 cells, and incubated at 37 • C, 5% CO 2 incubator and monitored daily for CPE.…”

Section: Beta-propiolactone Treatmentmentioning

confidence: 99%

Propagation, Inactivation, and Safety Testing of SARS-CoV-2

Jureka

Silvas

Basler

2020

Viruses

118

View full text Add to dashboard Cite

In late 2019, a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in Wuhan, the capital of the Chinese province Hubei. Since then, SARS-CoV-2 has been responsible for a worldwide pandemic resulting in over 4 million infections and over 250,000 deaths. The pandemic has instigated widespread research related to SARS-CoV-2 and the disease that it causes, COVID-19. Research into this new virus will be facilitated by the availability of clearly described and effective procedures that enable the propagation and quantification of infectious virus. As work with the virus is recommended to be performed at biosafety level 3, validated methods to effectively inactivate the virus to enable the safe study of RNA, DNA, and protein from infected cells are also needed. Here, we report methods used to grow SARS-CoV-2 in multiple cell lines and to measure virus infectivity by plaque assay using either agarose or microcrystalline cellulose as an overlay as well as a SARS-CoV-2 specific focus forming assay. We also demonstrate effective inactivation by TRIzol, 10% neutral buffered formalin, beta propiolactone, and heat.

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Gas chromatography-mass spectrometry method for determination of β-propiolactone in human inactivated rabies vaccine and its hydrolysis analysis

Cited by 19 publications

References 10 publications

Characterization of binding interactions of SARS-CoV-2 spike protein and DNA-peptide nanostructures

Characterization of binding interactions of SARS-CoV-2 spike protein and DNA-peptide nanostructures

Propagation, inactivation, and safety testing of SARS-CoV-2

Propagation, Inactivation, and Safety Testing of SARS-CoV-2

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