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Assays & Technologies

Respiratory Viruses Research Laboratory

Influenza GFP assay [Code No: IPK0301]

Using a recombinant influenza A virus expressing the green fluorescence protein (GFP) gene, infection of susceptible host cells is determined by measuring the increase in fluorescence intensity as a result of influenza infection. This is a rapid cell-based infection assay that can assess antiviral efficacy and is used primarily for high-throughput screening.

Influenza GFP imaging assay [Code No: IPK0302]

This is an image-based assay that visualizes the expression of green fluorescence protein (GFP) in cells infected with a recombinant influenza A virus harboring the GFP gene. The assay combines confocal fluorescence imaging technology and in-house image mining software capable of quantifying the percentage of infection in a live cell model, as well as other valuable information.

High-throughput microneutralization assay [Code No: IPK0303]

This assay is used for identifying virus neutralizing antibody species or antibody fragments from combinatorial antibody libraries in a live cell model. Utilizing confocal fluorescence imaging technology, infection is visualized by detection of the viral protein in the host cell, and quantified using our in-house image mining software. The virus neutralizing property is assessed by the reduction in the proportion of virus-infected cells.

Small interfering (si)RNA screening [Code No: IPK0304]

RNAi technology is an advanced molecular biology tool used to identify the roles of various cellular factors in successful viral replication. Gene knockdown using small interfering (si)RNA in a high-throughput system enables us to investigate the whole human genome or specific gene families for their involvement in the viral replication process.

Cytopathic effect (CPE) determination assay [Code No: IPK0305]

This general cell-based assay is used to assess cell death brought about by viral infection or compound toxicity. Cytopathic effects caused by infection with certain viruses, including influenza viruses, result in the loss of cellular metabolic activity (e.g., ATP production, oxidoreductase production) and cellular trafficking (e.g., lysosomes). These can be measured by metabolic conversion of reporter molecules such as oxyluciferin, resazurin, tetrazolium or lysosomal uptake of neutral red.

Viral entry assay [Code No: IPK0306]

This is a series of phenomic-based assays designed to investigate the different phases involved in the entry of viruses into the host cell. It is a high-content assay used to identify which stage of the viral entry process is blocked by treatment with an antiviral agent or knockdown of a critical host factor by small interfering (si)RNA.

Protein thermal stability shift assay [Code No: IPK0307]

This in vitro assay predicts the putative interaction of a ligand (e.g., small molecules) with the target protein. A shift in the melting temperature (thermal denaturation) of the protein in the presence of the small molecule indicates protein-ligand interaction, resulting in a change in structural integrity. Differential scanning fluorimetry measures the fluorescence emission of a dye that incorporates into the hydrophobic pockets of the protein during its transition into its denatured state.

Electrophoretic mobility shift assay (EMSA) [Code No: IPK0308]

This in vitro assay determines the inhibitory properties of small molecules against specific protein-ligand interactions by comparing the electrophoretic migration of the protein or ligand in a polyacrylamide matrix when separated or in a complex.

Molecular biology and virological laboratory techniques [Code No: IPK0309]

Classical virological laboratory techniques (e.g., virus amplification, titration) and advanced molecular biology techniques (e.g., real-time PCR, in vitro transcription assay) are used to investigate the biology of viruses in vitro, as well as in in vivo animal models.

Reverse Genetics (RG) system [Code No: IPK0310]

This advanced molecular biology tool allows the rescue of viruses through transcription of the genome and expression of essential viral proteins for replication in eukaryotic cells. In addition, genetic manipulation to generate specific mutations can be easily carried out using the plasmid constructs containing the viral genome segment(s).

Neuraminidase (NA) inhibition assay [Code No: IPK0311]

This is one of the in vitro assays designed to evaluate the putative mode of antiviral activity of potential anti-influenza compounds. The NA inhibition assay tests compounds for inhibitory properties against the enzymatic function of the influenza virus neuraminidase (NA) protein.

NF-κB (p65) nucleus translocation assay [Code No: IPK0312]

This is an image-based monitoring system used to visualize the activation of the NF-κB signaling pathway in vitro. The activated NF-κB is translocated from the cytoplasm to the nucleus. A GFP-fused NF-κB overexpressing cell line makes it possible to visualize the NF-κB activation.

NF-κB/GFP-Luc reporter assay [Code No: IPK0313]

This in vitro assay is specifically designed for monitoring the NF-κB signal transduction pathway. This technology is used for quantitative transcription activation reporter assays by the detection of GFP fluorescence as well as luciferase.

Interferon-Stimulated Response Element (ISRE)/GFP-Luc reporter assay
[Code No: IPK0314]

This in vitro assay is specifically designed for monitoring the interferon-induced signaling pathway. This reporter system responds to type I interferons (IFN-α and -β) that mediate signaling through STAT1 and STAT2 components of the JAK/STAT-signal transduction pathways. This technology is used for quantitative transcription activation reporter assays by the detection of GFP fluorescence as well as luciferase.