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Basic Characteristics of Mutations
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Mutation Site
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D614G |
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Mutation Site Sentence
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We have conducted a series of improvements,including using a C-terminal truncated form of spike protein and a D614G mutated spike. |
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Mutation Level
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Amino acid level |
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Mutation Type
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Nonsynonymous substitution |
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Gene/Protein/Region
|
S |
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Standardized Encoding Gene
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S
|
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Genotype/Subtype
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- |
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Viral Reference
|
-
|
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Functional Impact and Mechanisms
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Disease
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Cell line
|
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Immune
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- |
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Target Gene
|
-
|
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Clinical and Epidemiological Correlations
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Clinical Information
|
- |
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Treatment
|
- |
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Location
|
- |
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Literature Information
|
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PMID
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33521163
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Title
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Comprehensive and systemic optimization for improving the yield of SARS-CoV-2 spike pseudotyped virus
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Author
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Fu X,Tao L,Zhang X
|
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Journal
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Molecular therapy. Methods & clinical development
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Journal Info
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2021 Mar 12;20:350-356
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Abstract
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Virus neutralization assay is principally conducted by measuring the ability of the antibodies in patient sera to prevent the infection of susceptible cells by the virus. As SARS-CoV-2 is classified as a risk group 3 pathogen, neutralization assay using a live virus needs to be handled in a biosafety level 3 laboratory. To overcome this limitation, pseudotyped viruses have been developed as an alternative for the live SARS-CoV-2. However, one of the issues that we and others have encountered during the production of pseudotyped virus with SARS-CoV-2 spike protein was the low virus yield. In our own experience, we were only able initially to produce a stock with a virus titer that is more than two orders of magnitude lower than what we usually get with a vesicular stomatitis virus glycoprotein (VSV-G) pseudotyped lentiviral vector. We have conducted a series of improvements, including using a C-terminal truncated form of spike protein and a D614G mutated spike. Together, these have led to a significant improvement in the yield of the pseudotyped virus. Finally, our data show that using a high-affinity ACE2-expressing cell line resulted in a reduction in detection sensitivity of the neutralization assay.
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Sequence Data
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-
|