UV inactivation:
one wavelength,
wide use
The infectivity of the virus can be completely eliminated with ultraviolet radiation. Added value is that all this is done without chemicals and additives, in a few seconds.
Proteins are intact, RNA can be measured.
Advantages of the UV-C irradiation we use
0% infection
The virus is completely inactivated and can be treated in a normal microbiological environment. Checked with 5 different procedures.
intact proteins
There is no detectable difference in the amounts of active and UV-inactivated viral proteins. (ELISA)
Cellular processes identical to the original. (immunocytochemistry, flow)
even an unchanged nucleic acid
There is no significant change in the amount of RNA at the inactivation limit (RT-qPCR). After several irradiations, 1-2 Cq.
Virus cleaning
Virus cleaning procedure
With the further development of anion-exchange magnetic bead cleaning technology, we have created our own virus cleaning procedure, which allows for a high level of purity and significant recovery in addition to a 20x concentration of the virus isolate. In contrast to gradient centrifugation with an ultracentrifuge, which is considered the gold standard, the method takes 30 minutes. The loss is 40%. The recovery in terms of RNA quantity is 60-70% (RT-PCR). The purity index in terms of total protein is 96% (BCA assay). With transmission electron microscopy, we saw a picture typical of intact virions. The samples we examine are cellular supernatants, and in the case of a complex biological sample (e.g. blood serum), pretreatment of the sample may become necessary.
Advantages of our method:
can be done quickly (30 minutes)
results in adequate purity from a functional point of view (no cytopathy, cells can be examined, electron microscopic image is as expected)
can be optimized for maximum recovery/maximum concentration (75% or 11.5x concentration)
does not require significant equipment background (can be implemented in a laboratory with a simple desktop centrifuge)
We provided the developed virus cleaning technology to a university partner (University of Pannon) and to the PTE research group in the framework of scientific cooperation.
Immune response tests
Like most viral diseases, our body reacts to COVID-19 by producing antibodies and training special immune cells. Vaccines also want to create this immune reaction - just without the disease. When measuring the antibody immune response induced by the virus, in the traditional serological test (ELISA), only the antibody response to a fragment of one antigen of the virus is measured. On the other hand, our method is a new type of procedure: we can measure the set of antibodies formed against the entire virus and all its antigens. In this way, we can measure the processes that also take place in nature under laboratory conditions: since our immune system encounters the entire virus on the mucous membranes or in other tissues of our body. We also found a significant antibody response in the blood of people who did not know they had the disease and had not been vaccinated. Their blood could easily have provided the most excellent therapeutic blood product, yet neither they nor the doctors knew that it could be suitable. With the procedure, we can reveal hitherto unseen dimensions of the anti-coronavirus antibody response.
The cellular immune response induced by COVID-19 is also measured by capturing the crucial moment of natural immune processes. After blood collection, white blood cells are collected and mixed with the coronavirus itself. We use UV-inactivated coronavirus, which differs from the active virus in one important aspect: it is not contagious. However, its proteins are not damaged, so it behaves in the same way as the infectious virus. When mixed with white blood cells, the natural immune cells in our blood process the coronavirus at the cellular level, and then present it to professional immune cells - they almost ask: have you ever met the virus? Think about it: if our cells recognize the virus, we have a cellular immune response. The simplicity of the solution also includes the fact that the immune response to the entire virus is measured, completely tailored to the examined person.
The practical possibilities are numerous: it can be used to identify previously unknown infections, to select suitable donors in convalescent plasma therapy, or to measure the immune response to vaccines. It is important to note that our studies proved the raison d'être of the procedure in a clinical study with a moderate number of subjects. Therefore, in order to be widely used, it needs to be tested in a large number of clinical trials.