Vital Role In Viral Replication

In an attempt to increase the diagnostic possibilities of COVID-19 patients, this assay was developed for the detection of specific antibodies against one of the 16 non-structural proteins, the main viral protease (Mpro or 3CLpro), which plays a critical role in viral replication.

The Mpro protease plays a vital role in processing the polyproteins that are translated from the viral RNA. Although this protein is not exposed in the viral particle, Mpro is elicited after viral infection. Like other b-coronaviruses, SARS-CoV-2 is a positive-sense RNA virus that expresses multiple proteins as a single polypeptide chain, and Mpro cleaves the polyprotein to release mature proteins for the virus. Inhibitors that can block viral replication are promising potential drug candidates that could be used to treat patients suffering from the COVID-19 coronavirus infection.

If you are a particular and you want to acquire this test, please contact us in order to recieve a personalized proposal of distributors near you.

Our products are available through our extensive distribution network all over the world. Please contact us and we will give you all the information shortly.

You can place an order via email writing us to order@immunostep.com. Simply choose a kit and send us your order with the reference number stabished in the table.  You can also  contact us in +34 923 29 48 27 (Spain). 

CE/IVD kits are only suitable for clinical use in European Union and countries with EU conformity recognition agreements. Manufactured in Spain by Immunostep under CSIC Patent Licence.

Protocol: 

1. Add sample Incubation Add 100 µl of the positive control, the negative control, the calibrator and the prepared sample (100:1 dilution) into the individual wells of the microplate. It is recommended to use two wells per sample, including the controls and in particular the calibrator. Incubate for 60 minutes at +37°C. When the process is manual, cover the microplate with one of the protective foils provided.
2. Washing If necessary, remove the protective film. Empty the wells and then wash 4 times using 300 µl of 1X wash buffer in each wash. Leave the wash buffer in each well for 30 to 60 seconds per wash cycle. After washing, completely remove all liquid from the microplate by tapping it on absorbent paper with the openings facing downwards to remove all residual wash buffer.
3. Incubation of the conjugate Add 100 µl of the dilution (1:100) of the HRP-conjugated antibody to the wells and incubate for 30 minutes at +37°C. When the process is manual, cover the microplate with one of the protective films provided.
4. Washing If necessary, remove the protective film. Empty the wells and wash as described above (step 2).
5. Substrate incubation Add 100 µl of the chromogenic substrate solution (TMB) to each well of the microplate. Incubate for 10 minutes at room temperature (+18°C and +24°C) and protected from light.
6. Stopping Add 100 µl of the stop solution (1X – ready to use) to each well, trying to follow the same order in which the substrate solution was added.
7. Absorbance measurement Measure the optical densities (OD) of each well in a microplate spectrophotometer at 450 nm and a reference wavelength of between 620 and 650 nm within 30 minutes of adding the stopping solution.
8. Before measurement, carefully shake the plate to ensure an even distribution of the solution.

We strive to maintain sufficient inventory to meet your needs. In the rare case that a product is out of stock, you will be notified at the time of your order.