Language:

Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays

Viruses, 2020-05, Vol.12 (5), p.513 [Peer Reviewed Journal]

COPYRIGHT 2020 MDPI AG ;Attribution ;2020 by the authors. 2020 ;ISSN: 1999-4915 ;EISSN: 1999-4915 ;DOI: 10.3390/v12050513 ;PMID: 32384820

Full text available

Citations Cited by

Actions
1. Add to My Research
2. Remove from My Research
3. E-mail
4. Print
5. Permalink
6. Citation
7. EasyBib
8. EndNote
9. RefWorks
10. Delicious
11. Export RIS
12. Export BibTeX

Title:
Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays
Author: Crawford, Katharine H D ; Eguia, Rachel ; Dingens, Adam S ; Loes, Andrea N ; Malone, Keara D ; Wolf, Caitlin R ; Chu, Helen Y ; Tortorici, M Alejandra ; Veesler, David ; Murphy, Michael ; Pettie, Deleah ; King, Neil P ; Balazs, Alejandro B ; Bloom, Jesse D
Subjects: Angiotensin-Converting Enzyme 2 ; Antibodies ; Antibodies, Neutralizing ; Antibodies, Neutralizing - immunology ; Antibodies, Viral ; Antibodies, Viral - blood ; Antibodies, Viral - immunology ; Biochemical assays ; Biosafety ; Chemical tests and reagents ; Containment of Biohazards ; coronavirus ; Coronaviruses ; COVID-19 ; Diagnosis ; Enzyme-linked immunosorbent assay ; Health aspects ; HEK293 Cells ; Humans ; Infection ; lentiviral pseudotype ; Lentivirus ; Life Sciences ; Luciferase ; Medical research ; Medicine, Experimental ; Methods ; Microbiology and Parasitology ; neutralization assay ; Neutralization Tests ; Neutralization Tests - methods ; Peptidyl-Dipeptidase A ; Peptidyl-Dipeptidase A - metabolism ; Plasma ; Plasma - immunology ; Protocol ; SARS-CoV-2 ; Spike ; Spike Glycoprotein, Coronavirus ; Spike Glycoprotein, Coronavirus - analysis ; T cells ; Viral antibodies ; Virology ; Virus diseases
Is Part Of: Viruses, 2020-05, Vol.12 (5), p.513
Description: SARS-CoV-2 enters cells using its Spike protein, which is also the main target of neutralizing antibodies. Therefore, assays to measure how antibodies and sera affect Spike-mediated viral infection are important for studying immunity. Because SARS-CoV-2 is a biosafety-level-3 virus, one way to simplify such assays is to pseudotype biosafety-level-2 viral particles with Spike. Such pseudotyping has now been described for single-cycle lentiviral, retroviral, and vesicular stomatitis virus (VSV) particles, but the reagents and protocols are not widely available. Here, we detailed how to effectively pseudotype lentiviral particles with SARS-CoV-2 Spike and infect 293T cells engineered to express the SARS-CoV-2 receptor, ACE2. We also made all the key experimental reagents available in the BEI Resources repository of ATCC and the NIH. Furthermore, we demonstrated how these pseudotyped lentiviral particles could be used to measure the neutralizing activity of human sera or plasma against SARS-CoV-2 in convenient luciferase-based assays, thereby providing a valuable complement to ELISA-based methods that measure antibody binding rather than neutralization.
Publisher: Switzerland: MDPI AG
Language: English
Identifier: ISSN: 1999-4915
EISSN: 1999-4915
DOI: 10.3390/v12050513
PMID: 32384820
Source: Geneva Foundation Free Medical Journals at publisher websites
AUTh Library subscriptions: ProQuest Central
Hyper Article en Ligne (HAL) (Open Access)
MEDLINE
PubMed Central
Directory of Open Access Journals
ROAD: Directory of Open Access Scholarly Resources

Back to results list


INSPIRE LIBRARY - TON DUC THANG UNIVERSITY	(84-028) 37 755 057	Feedback
19 Nguyen Huu Tho St. Dist.7, HCM	thuvien@tdtu.edu.vn	Feedback

Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays

COPYRIGHT 2020 MDPI AG ;Attribution ;2020 by the authors. 2020 ;ISSN: 1999-4915 ;EISSN: 1999-4915 ;DOI: 10.3390/v12050513 ;PMID: 32384820

Searching Remote Databases, Please Wait