Powerful new option: QIAGEN ESEQuant LR3 and BioPorto gRAD
Improved development of rapid, lateral flow-based tests.
The generic Rapid Assay Device - gRAD - is a quantifiable, ready-to-use and generic lateral flow platform designed to speed-up the development of your own rapid assays. No specific antibodies are immobilized on the strip, making the test of a variety of biomarkers and labeling systems much more flexible and easier.
The ESEQuant LR3 Lateral Flow System is a comprehensive solution for the development of your own lateral flow test. A wide range of strips with either fluorescence or colorimetric labels or different number of strips and lines can be easily analyzed, which significantly speeds up the whole development process. Discover more on www.lateralflowreader.com.
As an example, a gRAD-based assay for quantification of neutrophil gelatinase-associated lipocalin (NGAL), a marker for acute kidney injury, was performed in an automated fashion on the LR3 system. In a first step an appropriate test method was set up utilizing the Lateral Flow Studio Software that is provided with the ESEQuant system.
The verification trial showed that the excellent dynamic range and sensitivity of the NGAL assay was well supported by the superior LR3 system’s properties. A direct comparison of LR3-based versus visual read-out resulted in an excellent concordance between both read-out modi. Thus, we conclude that both technologies, QIAGEN ESEQuant LR3 and Bioporto gRAD, provide a powerful and versatile toolbox for rapid and straightforward development of rapid, lateral flow-based tests.
Chart 1: Signal intensities on LR3 reader dependent on NGAL concentration on gRAD teststrip
"BioPorto's gRAD OneDetecion Strips provide a very convenient diagnostic platform that is extremely well suited for assay development. The capabilities of the gRAD strips allow the flexbility needed to solve unexpected issues encountered during optimization and associated gRAD reagents have proven to be extremely effective and were invaluable in completing a time-sensitive project. After experiencing major problems with lateral flow due to the matrix effects of biofluids, BioPorto's Dilution Buffer immediately resolved the issue and allowed us to produce our diagnostic."
For research use only - not for diagnostic procedures Brent McGill and Rhoel Dinglasan
Johns Hopkins Bloomberg School of Public Health
W. Harry Feinstone Department of Molecular Microbiology & Immunology
See Technical Note for further information.
With the new generic Rapid Assay Device (gRAD) from BioPorto Diagnostics it is now much easier and faster to develop your own customized qualitative or semi quantitative* lateral flow assay.
The gRAD OneDetection strip is an optimized generic lateral flow strip with a test line where a biotinylated capture antibody (or other biotinylated proteins) will bind, and a control line that will capture any antibody from mouse, rabbit and goat.
To generate your own specific gRAD OneDetection test, you just need to follow these simple steps:
When the sample of interest is added to the antibody solution, the antibodies then react and can form a complex with the analyte in the sample. The mixture will - if a complex is formed - now contain both free capture antibody and an antigen-antibody complex with the analyte.
*For the possibility of a quantitative option please contact us.
When the gRAD OneDetection strip is dipped into the mixture with the sample - migration along the strip occurs. The antibodies-sample complex mixture will flow by capillary force over the test and control lines. The biotinylated capture antibody - free and in complex - will bind to the test line. Uncomplexed gold detection conjugate will bind to the control line. After an assay-specific incubation time - typically between 10 and 15 minutes - the test line will be saturated and a stable intensity can be observed.
The color intensity of the test line depends on the concentration of the analyte.
BioPorto Diagnostics' unique patented gRAD OneDetection strip enables you to create a custom lateral flow assay specific to your needs – as the gRAD OneDetection strip is not analyte dependent.
You only need to provide:
** Sample material should be liquid in the form of urine, serum, plasma, drinking water or solid material dissolved in sample diluent.
The development of an assay based on the gRAD OneDetection strip consits of three steps:
After optimization of the capture and detection reagents, the developed test can be run using the gRAD OneDetection strip.
At BioPorto Diagnostics, we have a long history of listening to the needs of our customers and working with them to find a solution that fulfills their requests. So please feel free to contact us, if you have specific requirements or ideas that you would like to develop in collaboration with us. We are able to set up a solution to meet your needs.
Please fill our your details here.
In order to develop your own gRAD assay, analyte specific matched antibody pairs - a capture and a detection antibody is needed.
BioPorto Diagnostics has a long history of identifying and launching antibodies within highly specific areas. This has enabled us to offer a comprehensive range of matched antibody pairs.
Hereby BioPorto allows researchers to create their own customized and strong immunoassay solutions.
If you are looking for antibodies for your gRAD assays, please see our portfolio here.
How long does it take to develop my own assay using a gRAD OneDetection Kit?
To develop your assay, you need a suitable pair of antibodies – a biotinylated capture antibody and a detection antibody to be conjugated with colloidal gold. Both the capture and detection antibodies should specifically recognize the analyte you want to detect. For already existing antibody pairs, you should be able to have your first working test in the first day, optimization of the conjugation with gold and development of the assay on gRAD should take around 2 weeks, depending on the experience of the user.
Which size of gold particles should I choose for my gRAD assays?
The diameter of the colloidal gold particles may affect the sensitivity of the assay. Usually, the 40 nm particle in more sensitive that the 20 nm particle. On the other hand, smaller particles have a larger surface and require more antibody to coat, making it more robust to hook effects. We recommend the use of the 20 nm and 40 nm particles for comparison, and then choose for further development the size that gives the better results for your specific assay.
Can I use polyclonal antibodies with the gRAD OneDetection strips?
Yes. However, all polyclonal antibodies must be affinity purified with the antigen of interest. The use of antisera and protein A and G purified antibodies will result in a poor test response.
Can I use whole blood samples with gRAD OneDetection strip?
The current version of gRAD OneDetection strip is optimized to be used with liquid samples in the form of urine, serum, plasma, drinking water or solid material dissolved in the sample diluent. To use whole blood we recommend preliminary sample preparation. Contact us if you would be interesting in using whole blood or other sample matrices.
Can I use gRAD OneDetection strip with oligonucleotides?
The current version of the gRAD OneDetection strip have not been validated for use with oligonucleotides. The current gRAD OneDetection strip configuration is expected to be compatible to detect PCR reaction products were one primer is biotinylated and the other primer contains another tag such as a fluorescent label, DNP or DIG with a specific antibody conjugated to gold, latex or other detection systems.
Can I use gRAD OneDetection kit to develop a quantitative assay?
Yes. There is the possibility of developing quantitative assays with gRAD OneDetection kit. For more information and options on how to develop a quantitative assay with gRAD, please contact us here.
Can BioPorto Diagnostics help me find the right matched antibody pairs for my assay?
Yes. BioPorto has a portfolio of high-quality matched antibody pairs consisting of a biotinylated antibody and an antibody that can be conjugated with gold. If you cannot find the antibodies you need in our matched antibody pairs list, please let us know.
Does BioPorto Diagnostics provide custom services in assay development with gRAD OneDetection kit?
We have a long history of listening to our customers and addressing their needs. If you have any special requirements for developing your assays, we will be happy to collaborate with you and set up a solution that meets your needs.
Does BioPorto Diagnostics offer gold-conjugated antibodies?
At the moment BioPorto Diagnostics does not have a portfolio of readily available gold-conjugated antibodies, but feel free to contact us if you need help in the conjugation of antibodies to gold.
Can I apply for the gRAD Partner Program?
The gRAD Partner Program is meant for high-impact academic institutions or research centers trying to develop assays based on gRAD OneDetection kit. The Program offers a collaborative platform between your institution and BioPorto Diagnostics in the research and development of new rapid assays. Feel free to contact us to know which conditions apply.