A new method for the fast detection of a key antiviral
Interferons are proteins that are an important part of our natural defense systems. These proteins also exhibit remarkable antiviral activity. The recombinant human interferon α2b (rhIFNα2b) was approved by the US Food and Drug Administration in 1986. It has since been used as an antiviral agent for the treatment of hepatitis B and hepatitis C. Despite its widespread applications, a problem remains: the detection of rhIFNα2b is tedious and time-consuming.
Against this background, researchers from China recently developed a new method for the rapid and efficient detection of rhIFNα2b in a new study. This paper was made available online on July 8, 2021 and published in the Pharmaceutical Analysis Journal on April 30, 2022.
To develop their new method, they immobilized a new nanobody on a paper strip. The nanobody used in this method originally came from an alpaca, a species of the South American camelid mammal. It was then generated in the research lab using recombinant DNA technology – a technique used to subclone DNA fragments to obtain large amounts of synthetic proteins. This is usually accomplished using bacteria or other prokaryotic cells. A nanobody is a functional fragment of a larger antibody. Since the immobilized novel nanobody binds rhIFNα2b tightly and with high specificity, it was used for rapid and foolproof detection of rhIFNα2b.
According to Dr. Junzhi Wang: “Due to the advantages of nanobodies in terms of conservation, production and reagent cost, the lateral flow immunochromatography assay using nanobodies has great potential to replace traditional antibody-based ligand binding assays for rapid identification test of recombinant protein therapies.”
The research team characterized the binding for the I22-rhIFNα2b interaction, ie, binding between nanobody 122 and rhIFNα2b, using an Octet platform. The data obtained clearly indicated a tight bond. The binding specificity was further validated using Western blotting, a technique used to detect proteins using protein-specific antibodies.
“The rhIFNα2b products currently available in China include injections, injection powders, gels and pastes. The immunochromatography strips can only be used to assess liquids or powder products that can be dissolved and applied to the strips. diffusing along the strip via capillary action, gels and pastes do not meet this requirement,” explains Dr. Wang.
Interestingly, the developed rhIFNa2b detection assay has a detection limit of 1 µg/ml, which is lower than the existing limits. This makes it a more sensitive laboratory-based technique for rapid identification of rhIFNα2b. Another major advantage is the use of nanobodies for protein detection. This is because nanobodies can be obtained economically by harvesting cheap bacterial cells. In addition, large volumes of nanobodies can be obtained relatively easily using routinely used laboratory techniques.
dr. Wang says: “The operating time of rhIFNα2b identification was shortened from two days to several minutes with our assay. It can therefore meet the need for rapid detection of this family of recombinant protein products on the market and provide a good basis for improving the efficiency of the market counterfeit detection. In the future, rapid detection could be performed in an all-round way.”
In summary, the newly developed method could pave the way for smoother, faster and more accurate detection of recombinant or artificially generated proteins, enabling early diagnosis and treatment of hepatitis.
Xi Qin et al, Development of Novel Nanobody-based Lateral Flow Immunochromatographic Stripping Assay for Rapid Detection of Recombinant Human Interferon α2b, Pharmaceutical Analysis Journal (2021). DOI: 10.116/j.jpha.2021.07.003
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Quote: A New Method for the Rapid Detection of an Important Antiviral Agent (2022, June 9,), retrieved June 9, 2022 from https://phys.org/news/2022-06-method-fast-key-antiviral.html
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