RAPPID

RAPPID performs homogeneous ratiometric bioluminescent immunoassays by analyte-induced complementation of split NanoLuc luciferase for quantitative detection of proteins and antibodies.

Key Features:

• Homogeneous mix-and-measure assay: Performs immunoassays without separation or multiple wash steps.
• Ratiometric bioluminescent readout: Produces a ratiometric signal through combined reporter and calibrator luciferases for quantitative measurements.
• Split NanoLuc complementation: Uses analyte-induced complementation of split NanoLuc luciferase fragments to generate signal upon target binding.
• Protein G-mediated photoconjugation: Photoconjugates NanoLuc fragments to antibody sandwich pairs via protein G adapters.
• Calibrator luciferase: Incorporates a calibrator luciferase to enable in-sample calibration and correct for sample-related variability.
• Compatibility with complex media: Enables quantitative measurements directly in complex media such as blood plasma.
• Low-picomolar sensitivity: Achieves sensitive detection at low picomolar concentrations.
• Digital camera detection: Generates bioluminescent output that can be detected using a basic digital camera.
• Elimination of external calibration and multiple incubations: Reduces the need for external calibrators and multiple washing/incubation steps typical of heterogeneous immunoassays.
• Target analytes: Applicable to protein biomarkers, anti-drug antibodies, therapeutic antibodies, SARS-CoV-2 spike proteins, and anti-SARS-CoV-2 antibodies.

Scientific Applications:

• Protein biomarker quantification: Quantitative detection of protein biomarkers in biological samples.
• Anti-drug antibody monitoring: Detection and quantification of anti-drug antibodies in patient samples.
• Therapeutic antibody detection: Measurement of therapeutic antibody levels.
• SARS-CoV-2 antigen and serology testing: Detection of SARS-CoV-2 spike protein and corresponding anti-SARS-CoV-2 antibodies.
• Assays in complex media: Quantitative assays directly in blood plasma and other complex sample matrices.

Methodology:

Analyte-induced complementation of split NanoLuc luciferase fragments photoconjugated to antibody sandwich pairs via protein G adapters, combined with a calibrator luciferase to produce a ratiometric bioluminescent signal detectable by a digital camera.