Protein A ELISA Protocol 10-29-20 Version 1
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Protein A ELISA Protocol 10-29-20.docx
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Created: 22nd Nov 2023 at 23:09
Last updated: 3rd Jan 2024 at 17:38
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Version 1 (earliest) Created 22nd Nov 2023 at 23:09 by John Cornelius
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https://orcid.org/0009-0003-6381-9603
Public web page: Not specified
Submitter: John Cornelius
Studies: Viral Protein Accumulation of Zika Virus Variants Links with Regulation ...
Assays: Bioinformatics analyses of NanoString data, Deep sequencing of ZIKV stocks and sequence alignment, FFU assay for determining the titers of ZIKV stocks, FFU assay for measuring viral spread, Flow cytometry, Immunoblot analysis, Immunofluorescence analysis, Plaque assay, Protein lysate quantification, Quantification of extracellular and intracellular RNA, Quantification of extracellular and intracellular virions, Sample - Metadata, Transcript analysis by NanoString, Transcript analysis by SYBR green qRT-PCR, Viral copy number quantification by TaqMan qRT-PCR, ZIKV attachment and entry assay, ZIKV infection, ZV-13 antibody purification and quantification
Asian lineage Zika virus (ZIKV) strains emerged globally, causing outbreaks linked with critical clinical disease outcomes unless the virus is effectively restricted by host immunity. We have previously shown that retinoic acid-inducible gene-I (RIG-I) senses ZIKV to trigger innate immunity to direct interferon (IFN) production and antiviral responses that can control ZIKV infection. However, ZIKV proteins have been demonstrated to antagonize IFN. Here, we conducted in vitro analyses to assess ...
Submitter: John Cornelius
Investigation: Zika Virus
Assays: Bioinformatics analyses of NanoString data, Deep sequencing of ZIKV stocks and sequence alignment, FFU assay for determining the titers of ZIKV stocks, FFU assay for measuring viral spread, Flow cytometry, Immunoblot analysis, Immunofluorescence analysis, Plaque assay, Protein lysate quantification, Quantification of extracellular and intracellular RNA, Quantification of extracellular and intracellular virions, Sample - Metadata, Transcript analysis by NanoString, Transcript analysis by SYBR green qRT-PCR, Viral copy number quantification by TaqMan qRT-PCR, ZIKV attachment and entry assay, ZIKV infection, ZV-13 antibody purification and quantification
Hybridoma supernatant was harvested as the source of ZV-13 antibody once ~95% cell death was observed. The supernatant was clarified by centrifuging for 10 min at 400 × g at 4°C and then filtered with a 0.22-μm filter prior to antibody purification via protein A affinity chromatography using the Pierce Protein A IgG purification kit and NAb Protein A Plus spin columns (all from Thermo Fisher-Life Technologies). Purified ZV-13 antibody was concentrated and buffer-exchanged into 1× phosphate-buffered ...
Submitter: John Cornelius
Assay type: Immunoassay
Technology type: Affinity Chromatography
Investigation: Zika Virus
