RAPPID enabling performant, fast, easy to use Homogeneous Immunoassay in less than 30 minutes
Currently, the standard method for bioanalysis of antibodies is by means of immunoassays to monitor the concentration of an antibody or antigen in the blood stream.
Heterogeneous immunoassays such as ELISA have become indispensable in modern bioanalysis, yet translation into point-of-care assays is hindered by their dependence on external calibration and multiple washing and incubation steps.
ELISA assays require a trained technician to undertake a series of carefully timed wash and addition steps and are also time-consuming.They also require novel anti-idiotypic antibodies which are not always available and often very challenging to produce.
LUMABS BV introduces RAPPID (Ratiometric Plug-and-Play Immunodiagnostics) technological platform, as a comprehensive platform of ratiometric bioluminescent sensors that can be readily adapted to suit a very broad range of biomolecular targets. a mix-and-measure homogeneous immunoassay platform that combines highly specific antibody-based detection with a ratiometric bioluminescent readout.
RAPPID is based on the complementation of antibody-conjugated split NanoLuc® luciferase to detect the formation of a sandwich immunocomplex in solution with a high intrinsic signal-to-background output. RAPPID does not require additional protein engineering and uses a patented protein conjugation method to generate well-defined antibody conjugates directly from available, out-of-shelf antibodies.
The patented innovation introduces an internal calibrator, a green light-emitting luciferase as an innovative approach to provide a robust blue-over-green ratiometric readout and direct internal calibration. The ratiometric signal of RAPPID sensors is stable over time and less sensitive to experimental conditions such as temperature and substrate concentration, effectively addressing this fundamental problem of intensiometric bioluminescent assays.
LUAMBS BV develops assay-specific RAPPID sensors that allow low-picomolar detection of several protein biomarkers, anti-drug antibodies, therapeutic antibodies, and viral spike protein and anti-viral antibodies. Development and implementation of a typical RAPPID assay for a new biomolecular target is straightforward and consists of three steps:
(1) the selection of a pair of (already available) antibodies that bind the target analyte
(2) crosslinking of the antibodies to the protein luciferase adapters using a proprietary protocol
(3) addition of both antibody-luciferase conjugates, the calibrator luciferase and the NanoLuc substrate to the sample, followed by detection of the emission ratio of blue over green light
With its easy-to-implement standardized homogeneous workflow, RAPPID provides an attractive, fast, and cost-competitive alternative to traditional immunoassays in clinical laboratories, academic research, BioPharma drug screening and for point-of-care applications.
The ratiometric luminescent signal can be recorded in a one-step assay in real-time with any commercially available Luminometer or even with a standard digital camera, highlighting the potential for point-of-care testing.
In the case of antibody detection, the dynamic range of the assay and its scope can be further increased with an evolved set of RAPPID variants that use genetic fusions of the split luciferase components to the antibody’s antigen. The broad scope and excellent analytical performance of RAPPID have been demonstrated by developing assays for a range of clinically relevant biomarkers, including cardiac troponin I, C-reactive protein (CRP), three anti-drug-antibodies (ADAs), and two therapeutic antibodies, displaying robust increases in emission ratio of up to 36-fold.
RAPPID works directly from 1- μL patient plasma samples
Another patented technology is the Lumabs (Luminescent Antibody Sensor) approach that uses our proprietary methodology based on a specific reagent for each antibody monitored. Our method negates the need for complex assay protocols, wash steps and can be achieved in a much-reduced timeframe, ensuring data is available in a significantly more convenient and efficient timeframe providing benefits for patients, clinicians, and laboratory analysts alike.
Lumabs Technology reagents consist of a light-emitting enzyme covalently attached, via a semi-flexible linker, to a fluorescent acceptor. Typically we use a blue-light luciferase and a green fluorescent protein. In the absence of the specific antibody in question, the light-emitting and the fluorescent elements are kept in close proximity by means of a pair of weakly interacting binding partners. In the presence of the antibody in question, tailored elements within the Lumabs Technology sensor bind respective Fab regions of the antibody so disrupting the close alignment of the light-emitting and fluorescent components. This conformational change is designed to result in a loss of fluorescent emission and an increase in the shorter wavelength light from the un-paired light-emitting enzyme.
Ratiometric analysis of the emitted light (e.g. blue versus green) enables use in Bioanalysis and TDM in complex samples, such as plasma, and is self-correcting for substrate fluctuations. Lumabs Technology proprietary technology thus provides unprecedented accuracy and ease of use in Bioanalysis, TDM and related applications.
The nature of the Lumabs Technology reagent enables simple to mix and measure assays to be performed rapidly and without the need for sequential wash and addition processes (as compared to ELISA based methods). Indeed, Lumabs Technology are suited to complete automation use in a point of care or even home testing setting with the potential to enable self-administration of the exact required amount of therapeutic, at the optimal time and so optimize therapeutic coverage.
We have also developed a variety of different configurations of our technology for a range of applications. We continue to further advance the technology in order to provide enhanced solutions for a range of needs.