Coronavirus SARS-CoV-2 (COVID-19) Vaccine Research

Cell count/cell viability

Cell counts and cell viability are fundamental assessments in numerous biological experiments, including many virology applications such as viral titer and infectivity. Cell microscopy is commonly used to capture brightfield and fluorescence images of cells in order to count them and assess whether live/dead, respectively.

Here’s a few application notes on cell counting and cell viability to aid in your COVID-19 vaccine research.

• Live/Dead (EarlyTox™ Live/Dead Assay Kit) staining - Measuring cell viability and cytotoxicity with the EarlyTox Cell Integrity Kit
• MTT assay - Assess cell viability and proliferation with colorimetric readouts
• Transfection efficiency - Optimizing GFP transfection with the SpectraMax i3 Multi-Mode Detection Platform
• Cell proliferation (Cell growth) - Confluence and growth curve generation on the CloneSelect Imager
• Cell viability (CellTiter-Glo® Luminescent Cell Viability Assay) - Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader

Cell line development

Cell line development requires the discovery of single cell-derived clones that produce high and consistent levels of the target therapeutic protein with desired quality attributes, possible to be scaled up for commercial GMP production.

Here’s a few application notes on cell line development to aid in your COVID-19 vaccine research.

• Colony picking - Enhanced development of virus-specific hybridomas using ClonePix and CloneSelect Imager Technologies
• Monoclonality verification - Confident assurance of clonality using calcein AM with minimal effect on viability
• Cell sorting - A workflow combining single-cell isolation and microplate imaging improves cloning efficiency with higher assurance of clonality
• Cell line knock-ins/outs - Validate CRISPR-Edited Cells using Imaging and Western Blot Detection on a Microplate Reader

COVID-19 research solutions

Molecular Devices is committed to supporting scientists that are researching COVID-19 cellular response and vaccine development by offering technology and solutions that you can deploy rapidly. We have validated and compliant laboratory solutions including microplate readers, washers, biopharma and cellular imaging systems to meet your all your research needs.

Go to COVID-19 Research  

IgG Quantification

In therapeutic protein engineering and cell line development, IgG quantification is the process of measuring the amount of immunoglobulin G (IgG), a common class of therapeutic proteins, produced by a genetically modified cell line. This is important for evaluating and monitoring the productivity of the cell line and selecting the best candidate for further development of therapeutic antibodies.

• High-throughput IgG quantitation platform for clone screening during drug discovery and development

• Poster: High-throughput IgG quantitation platform for clone screening during drug discovery and development

• Qualitative measurement of SARS-CoV-2 spike and nucleocapsid IgG antibodies in serum samples

• Virology and Vaccine Research: Microplate Reader Solutions

Luciferase assays

Luciferases are enzymes that use a substrate called luciferin, along with oxygen and ATP, in an energetic process that produces light—like the yellow glow of fireflies. The power of luciferase has been harnessed by scientists to devise reactions whose light output is used to monitor biological processes including gene expression, biomolecular binding, and cell viability.

Here’s a few application notes on luciferase assays to aid in your COVID-19 vaccine research:

• Measuring luciferase expression using the SpectraMax Glo Steady-Luc Reporter Assay Kit
• Dual-Luciferase Reporter (DLR) Assay
• Monitor NF-κB activation with a sensitive dual reporter assay
• Detect dual luciferase expression
• Highly sensitive dual luciferase detection with the SpectraMax DuoLuc Reporter Assay Kit

Protein and antigen binding affinity

Binding affinity is a measure of how strongly two molecules (often an antibody/antigen pair) interact with one another. Many different types of assays have been developed to assess binding affinity due to the significant heterogeneity amongst proteins including their size, location, solubility, and many other characteristics.

Learn more about the common applications for protein/antigen binding affinity to aid in your COVID-19 vaccine research:

• ELISA (IgG/IgM) - Enzyme-Linked Immunosorbent Assay (ELISA)
• Cell-based binding - Increase sensitivity in no-wash assays using confocal imaging
• HTRF- HTRF profiling services on SpectraMax iD5 Multi-Mode Microplate Reader
• AlphaScreen/AlphaLISA - AlphaLISA Screen on the SpectraMax Paradigm Multi-Mode Microplate Detection Platform

Protein and nucleic acid quantification (non-specific)

Experimental protocols involve a significant amount of assay/sample preparation, however, they also lengthen timelines in order to assure quality. Balancing the need for speed with the need for quality assurance can be accomplished with non-specific methods of quantification. Total amounts of proteins and/or nucleic acids can be measured using UV absorbance on a microplate reader.

Here’s a few application notes on protein and nucleic acid quantification (non-specific) to aid in your COVID-19 vaccine research:

• Colorimetric protein Assays (BCA/ Bradford Assays) - Measure total protein in cell lysates with SpectraMax ABS Plus Microplate Reader
• Protein quantification (using UV absorbance) - Direct protein quantitation using the SpectraDrop Micro-Volume Microplate
• DNA/RNA quantitation (using UV absorbance) - DNA and RNA absorbance measurements using SpectraMax Microplate Readers
• Fluorescence-based DNA quantification - Quantitation of dsDNA samples using the SpectraMax Quant dsDNA Assay Kits

Protein and nucleic acid quantification (specific)

Many biological experiments aim to alter protein expression within a cell, whether through genetic engineering or drug treatment. In order to understand the effect of such manipulation, it is critical to measure the response of specific protein(s). Antibodies are commonly used in bioassays (immunoassays) for this purpose due to their inherent ability to specifically bind to a protein of interest.

Learn more about the common applications for protein and nucleic acid quantification (specific) to aid in your COVID-19 vaccine research:

• ELISA (Cytokine secretion) - Enzyme-Linked Immunosorbent Assay (ELISA)
• Western blot - Western Blot
• HTRF (cytokine secretion) - Normalize HTRF cytokine assays to cell viability
• AlphaScreen/AlphaLISA - AlphaLISA Screen on the SpectraMax Paradigm Multi-Mode Microplate Detection Platform
• ValitaTITER (IgG quantification using fluorescence polarization) - Streamline the workflow for measuring IgG in cell line development

Vaccine development workflows

Vaccine development workflows vary depending upon the platform (e.g. inactivated virus vs. DNA vaccine) chosen, each having its own advantages. Here we've addressed a variety of virus-related workflows – from antigen and antibody discovery to cell line development.

Learn more about the common applications for vaccine development workflows to aid in your COVID-19 vaccine research:

• Antigen / Immunogen discovery workflow
• Antibody discovery - phage display workflow
• Antibody discovery - hybridoma screening workflow
• Stable cell line workflow

Viral neutralization

Vaccine and antiviral drug efficacies are assessed by their ability to inhibit viral entry (infection) into cells grown in culture.

• Plaque reduction neutralization assay (PRNT)
• Viral reduction neutralization assay (VRNT) - Virus Reduction Neutralization Test: A Single-Cell Imaging High-Throughput Virus Neutralization Assay for Dengue
• Microneutralization    Development of a High-Throughput Respiratory Syncytial Virus Fluorescent Focus-Based Microneutralization Assay

Viral pathogenesis (Viral pathways)

Understanding the basic mechanisms of viral entry, replication, shedding, etc. is critical for the development of effective treatments towards infectious diseases. Fluorescence microscopy is a fundamental tool in any virology laboratory because it allows for visualization of viral pathways at high resolution and over time. Cell signaling and reporter gene assays can be used to explore cellular pathways affected by viruses and are run using microplate readers.

Here’s a few application notes on viral pathogenesis (Viral pathways) to aid in your COVID-19 vaccine research:

• Internalization - Visualize subcellular vesicles to quantitate autophagy in neuronal cells
• Reporter gene assays - Measuring luciferase expression using the SpectraMax Glo Steady-Luc Reporter Assay Kit
• Cell signaling (GPCR) - Complete cAMP workflow solution using the CatchPoint cAMP Fluorescent Assay Kit

Viral titer

Viruses infect cells in order to replicate, a task which they cannot accomplish on their own. Measuring the amount of virus in a cell-free sample does not provide information regarding their infectivity, limiting its usefulness. Instead, cell imaging is used to detect the ability of a given virus to infect cells in culture. Viral concentrations are determined from the fraction of infected cells.

• Plaque assay - University of Zurich uses the ImageXpress Widefield System to investigate host pathogen interactions
• Focus forming assay
• Endpoint dilution