Therapeutic Development

Coronaviruses provide two attractive targets for small-molecule therapeutic intervention: the Papain-Like Protease (PLPro) and the 3C-like protease (3CL or “Main”).  Both enzymes are non-structural, cysteine proteases that are essential to the viral life cycle.  Because mammals lack proteases with similar substrate preferences it’s possible that protease inhibitors developed against these viral proteins will have low toxicity. R&D Systems featured key Protease Reagents for Coronavirus Research. 

Featured Protease Reagents for Coronavirus Research

R&D Systems offers a range of SARS-CoV-2 proteins and other Coronavirus proteins with the same industry-leading quality specifications as our other recombinant proteins. Our SARS-CoV-2 Spike proteins exhibit high affinity binding to human ACE-2 in both ELISA and SPR. And the Coronavirus proteases, Papain-like Protease and 3CL protease, are tested for bioactivity using biologically relevant fluorescent substrates. 

Receptor binding is the first step in viral infection. For coronaviruses, a cell-surface S protein mediates their entry into host cells. The S protein on SARS-CoV-2 recognizes and binds to ACE-2. Thus, both proteins could be viable therapeutic targets. Blocking the interaction between ACE-2 and the S protein, for example with a soluble S protein or an anti-ACE-2 antibody could, in theory, prevent SARS-CoV-2 infection. The development of COVID-19 therapeutics, though, is encumbered by the fact that SARS-CoV-2 exhibits high pathogenicity and infectivity and needs to be handled under biosafety level 3 conditions. To propel the development of COVID-19 therapeutics, we developed a flow cytometry-based, in vitro assay for evaluating the effectiveness of antibodies and/or small molecules to block the binding of the S protein to ACE-2.

ACE-2 Flow Cytometry Blocking Assay Principle. Cells are transfected with human ACE-2 DNA and express the receptor on their cell surface. When incubated with a recombinant His-tag SARS-CoV-2 Protein RBD, the protein will bind to ACE-2 (A). SARS-CoV-2 binding is detected with an APC-Conjugated His-Tag Antibody. Incubating the cells with an ACE-2 blocking antibody prior to the SARS-CoV-2 Protein RBD allows the ACE-2 antibody to bind to the ACE-2 receptor, blocking SARS-CoV-2 binding (B).

ATCC has provided stem cell resources to the research community for more than a decade, with a growing portfolio of cultures to choose from, including mouse embryonic stem cells, human embryonic stem cells, human mesenchymal stem cells (MSC), and human iPS cells. ATCC has developed complete cell culture solutions to support human MSCs and iPS cells in the collection. The MSCs can be used for studies of adult stem cell differentiation, regenerative medicine, and the creation of induced pluripotent stem cells.

Regarding exosomes, Isolation techniques are demanding; however, it is critical to understand the composition of the vesicles in your preparation. Biases can be introduced during sample preparation, such as: DNA, RNA, protein, and lipid extraction; PCR amplification; library preparation; sequencing; and/or data interpretation. To overcome these challenges, ATCC offers exosomes derived from well-authenticated cell lines and hTERT-immortalized Mesenchymal Stem Cells. These extracellular vesicles can be used as standards for creating diagnostic tests and studying exosome composition, epigenetic reprogramming of cells, and disease markers.

> Download the ATCC Stem Cell Solutions Brochure

> Download the ATCC Exosomes Brochure

Sexton is committed to providing tools to the cell and gene therapy industry that improve our customer’s outcomes and reduce risks. Stemulate® pooled human platelet lysate and nLiven PR, processed with pathogen reduction technology to further reduce risks, offer a strong history of use for the production of multiple cell-types to accomplish that objective.

Referenced in nearly 100 publications and presentations, human platelet lysate (Stemulate and nLiven PR^TM) have now been established as excellent growth supplements for MSCs derived from a  range of tissues including adipose, bone marrow, cartilage, dental pulp and umbilical cord.

We understand everyone’s research is unique. If you cannot find the solution you need for your MSC culture, or you need a specific media supplement, or if you’re interested in customizing a neutralising antibody, please contact our team at lifescience@invitro.com.au

Several research projects have been initiated to investigate the use of mesenchymal stem cells (MSCs) as a treatment option for the adverse effects caused by SARS-CoV-2 infection. Bio-Techne offers workflow solutions for MSC research. View the R&D Systems™ MSC resource page to see all of the products we have for the isolation and culture, verification, differentiation , and investigation of MSCs.

Additionally, using lung organoids to simulate the pulmonary environment can provide a more accurate evaluation of drug efficacy. Visit our Organoid Resource page on the R&D Systems website to view all of the reagents and technical resources we have, including webinars, protocols, and recipes, to help researchers build consistent lung organoid cultures.

Human mesenchymal stem cells were cultured in StemXVivo™ Mesenchymal Stem Cell Expansion Media (Catalog # CCM004) and differentiation was induced as indicated using the media supplements included in the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006). The kit also contains a Goat AntiMouse FABP-4 Antigen Affinity-Purified Polyclonal Antibody (adipocytes), a Goat Anti-Human Aggrecan Antigen Affinity-Purified Polyclonal Antibody (chondrocytes), and a Mouse Anti-Human Osteocalcin Monoclonal Antibody (Osteocytes) for the confirmation of differentiation status. The cells were stained using the NorthernLights™ 557-conjugated Donkey Anti-Goat (Catalog # NL001; red) or Anti-Mouse (Catalog # NL007; red) IgG Secondary Antibodies, and the nuclei were counterstained with DAPI (blue). All cited reagents are from R&D Systems.

> Download our new Tools to Support Coronavirus Research brochure. 

PRODUCTS FOR MSCS & PRODUCTS FOR LUNG ORGANOIDS are included in the Brochure.

Identifying compounds that induce toxicities is an important part of lead-optimization during drug discovery. However, most commonly used end‐point assays lack the sensitivity to capture many compounds with mitochondrial toxicity, leading to late‐stage failures.

Agilent Seahorse Bioanalyzer provide real-time, direct measurements of mitochondrial metabolism with the highest specificity and sensitivity. With these solutions, you can be confident the data you're collecting provides an accurate picture of mitochondrial toxicity risk enabling you to make informed decisions on your drug development pipeline.

> Download Application Note

Immune cell isolation and culture are necessary for both the basic research of immune cell functions during SARS-CoV-2 infection, as well as for expanding specific immune cell populations for therapeutic purposes. Bio-Techne is focused on providing the highest quality reagents for isolating, differentiating, and identifying immune cells.

PRODUCTS TO INVESTIGATE THERAPEUTIC MONOCLONAL ANTIBODIES

BIOSIMILAR ANTIBODIES

ANTI-IDIOTYPE ANTIBODIES

Research have shown that the CoV-2 spike (S) protein binds human ACE-2 located on the surface of mucosal cells, resulting in fusion of viral and cell membranes for viral entry. Viral entry also requires priming of the S protein by host cell proteases including TMPRSS2 and Cathepsin B/L. Involvement of ADAM17/TACE in this process, which cleaves ACE-2, remains questionable.

Since the S protein, ACE-2, TMPRSS2, and Cathepsin B/L, have important functions in the SARS-CoV-2 life cycle, these proteins serve as potential targets for drug development.

Bio-Techne offers various reagents to support the development of neutralizing antibodies and inhibitors against proteins involved in key steps in SARS-CoV-2 infection.

TMPRSS2 Products