Assays for HTS
Assays for High-Throughput Screening (HTS)
Quantibody: Multiplex ELISA Array
RayBiotech Quantitative Proteomics Services uses the Quantibody® glass slide array platform for simultaneous quantitative measurement of multiple cytokines, growth factors, proteases, soluble receptors, and other proteins in a wide variety of sample types.
Quantibody® is an array-based multiplex ELISA system for simultaneous quantitative measurement of multiple cytokines, growth factors, proteases, soluble receptors, and other proteins in a wide variety of sample types. Quantibody® combines the high specificity and sensitivity of ELISA with the high throughput of the glass chip-based array.
With this platform, only 50 µl of sample is needed for quantification of up to 40 proteins in quadruplicate, making this array 80 times more efficient than traditional ELISA.
Each 75mm x 25mm glass slide is spotted with 16 identical antibody arrays (also called “subarrays”). Within each subarray, the capture antibodies along with controls are spotted in quadruplicate. The slide comes with a 16-well removable gasket and chamber assembly that allows for simultaneous processing of up to 14 samples per slide. To achieve even higher throughput, 4 slides can be nested into a tray which matches a standard microplate, allowing for automated processing of 64 arrays simultaneously. The latest automated liquid handling workstations can accommodate several trays at once, allowing hundreds of samples to be processed per day.
Features and Benefits of Quantibody
- Less sample, more data: only 50 µl of sample is needed for quantification of up to 40 cytokines
- 4-hour processing time
- More cost-effective than ELISA
- No interference between capture antibodies (unlike bead-based multiplex assays)
- No dedicated equipment required
- Customizable – create a custom array from our list of targets
Contents of kit
- Glass Chip with antibody arrays
- Sample Diluent
- Lyophilized protein standard mix
- Detection antibody cocktail
- Streptavidin-Fluorescent dye
- Wash buffer
*Accessories include: 16-well incubation chamber with gasket, protective cover, snap-on sides, adhesive film
- High-throughput profiling of cytokine expression
- Validation of semi-quantitative antibody array results
- Identifying potential molecular targets for drug development
- Identifying the molecular mechanisms of drug action
- Identifying crucial factors involved in disease processes
- Discovering biomarkers for disease management
- Discovering expression patterns for molecular classification of diseases
RayBio® Protein Arrays are a versatile and innovative series of products featuring immobilized proteins spotted onto a glass slide support. Protein arrays allow a much broader view of protein activity than a traditional ELISA. RayBio arrays come in smaller, research specific panels as well as broad screening panels.
- Allergen Arrays
- Auto-Immunome Arrays
- Cancer Autoantibody Arrays
- SARS-CoV-2 (COVID-19) Arrays
- Peptide Mapping Arrays
Like a traditional sandwich-based ELISA, Quantibody uses a matched pair of antibodies for target protein detection. A panel of capture antibodies is printed in multiple identical arrays on a standard slide. After a blocking step, samples are incubated with the arrays. Nonspecific proteins are then washed off, and the arrays are incubated with a cocktail of biotinylated detection antibodies, followed by a streptavidin-conjugated fluor. Signals are then visualized using a fluorescence laser scanner.
To quantify target protein concentrations, the array-specific protein standards, whose concentrations have been predetermined, are provided to generate an 8-point standard curve of each target protein. By comparing signals from unknown samples to the standard curve, the unknown cytokine concentration in the samples will be determined.
G-Series: Glass Slide Antibody Arrays
Antibody arrays allow researchers to conduct rapid, accurate expression profiling of hundreds of cytokines, chemokines, growth factors, proteases, soluble receptors, and other proteins from any biological fluid.
RayBio® Glass Slide-based Antibody Arrays (G-Series) were introduced as a “second generation” array platform after the chemiluminescent membrane arrays (C-Series). The G-Series uses a 75mm x 25mm glass chip platform which delivers easier handling along with the use of much lower sample volumes. The glass slides also utilize a fluorescent signal readout, allowing a wider dynamic detection range than can be achieved by chemiluminescence.
G-Series Antibody Arrays are particularly well-suited for projects with very limited sample volumes (as little as 10 μl per array) and large numbers of samples. Dozens of arrays can be processed per day by a single technician. Furthermore, the slide chamber assembly is compatible with many automated arrays processing systems, allowing high-throughput studies to be performed with very little hands-on time.
Features of G-Series Antibody Arrays
- High sensitivity (pg/mL to ng/mL levels)
- Large dynamic scale of detection (4 orders of magnitude)
- Compatible with many sample types
- High throughput
- Low sample consumption: as little as 10 µl of original sample required per array
- Customizable – create a custom array from our list of targets
How It Works
Similar to the C-Series (membrane-based) arrays, G-Series Antibody Arrays utilize the sandwich-ELISA design principle. In this assay, capture antibodies are printed in multiple identical arrays on a standard-sized histology slide. After a blocking step, samples are incubated with the arrays. Nonspecific proteins are then washed off, and the arrays are incubated with a cocktail of biotinylated detection antibodies, followed by a streptavidin-conjugated fluor. Signals are visualized using a fluorescence laser scanner.
Comparison to traditional ELISA
- More Data, Less Sample: G-Series Antibody arrays provide high-content screening using less sample volume than ELISA. Only 50 µl needed (after dilution) to detect up to 40 different biomarker proteins. As little as 10 µl original sample!
- Global View of Cytokine Expression: Antibody array screening improves the chances for discovering key factors, disease mechanisms or biomarkers related to cytokine signalling.
- Greater Sensitivity: As little as 4 pg/ml of MCP-1 can be detected using the G-Series array format. In contrast, our similar MCP-1 ELISA assay has a sensitivity of 40 pg/ml of MCP-1.
- Increased Range of Detection: ELISA assays typically detect a concentration range of 100- to 1000-fold, however, RayBiotech arrays can detect IL-2 at concentrations of 25 to 250,000 pg/ml, a range of 10,000-fold.
- Better Precision: As determined by densitometry, the inter-array Coefficient of Variation (CV) of spot signal intensities is 5-10%, comparing favourably with ELISA testing (CV = 10-15%).
G protein-coupled receptors (GPCRs) represent the largest family of membrane proteins. Located on the surface of cells in the plasma membrane, GPCRs bind to a wide variety of ligands, such as hormones, neurotransmitters, lipids, nucleotides, and ions, and are critical intermediaries in the transmission of signals from the outside to the inside of the cell.
GPCRs are the largest family of cell-surface membrane receptors and one of the most intensely studied drug targets, hold out tremendous promise for scientists. In fact, therapies that target GPCRs include ligands from all pharmacological classes (agonists, antagonists, inverse agonists, allosteric modulators) used to treat diseases in every major organ system and for several therapeutic areas, including cardiovascular diseases, neurosciences, respiratory ailments, metabolic disorders, oncology, and rare diseases. What’s more, their surface location makes them accessible to a whole host of druglike molecules.
They are therefore of significant importance in several therapeutic areas, such as Cardiovascular diseases, Neurosciences, Respiratory diseases, Metabolic diseases, Oncology, and rare disease. It has been estimated that between one-third and one-half of all marketed drugs act by binding to GPCRs (475 FDA-approved drugs target 108 GPCRs).
There are kits to address the whole GPCR activation pathway including:
- Receptor binding (Ligand binding assay)
- Gs, Gi, Gq Signaling
- Beta-arrestin signaling
- Phosphorylation Pathways
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