Inhibitor Screening
To identify or confirm activity with the target.
BellBrook Labs
1232 Fourier Drive, Suite 115
Madison, Wisconsin 53717 USA
(608) 443-2400
info@bellbrooklabs.com
Innate Immunity – Products & Services
Unlike the adaptive immune response, the innate response is the part of the immune system that acts quickly and is considered the first line of defense against pathogens and foreign DNA, such as tumor cells. To be effective, the innate immune response relies on several types of immune cells and a multitude of proteins, such as interferons (INFs), which typically fight microbial infection-causing pathogens. Fighting unwanted microbes is essential, but dysregulation of the innate immune system leads to many inflammatory and autoimmune disorders. BellBrook Labs is leveraging its Transcreener technology to build enabling products and services focusing on targets found to help regulate innate immunity. Our goal is to help accelerate the discovery of potential treatments for autoimmune disease and cancer.
Full-Length Human cGAS Titration
Transcreener cGAS Activity Assay
When free DNA is in the cytoplasm of eukaryotic cells, it can be an indication of damage or infection to the cell. This DNA triggers an immune response led by type I interferon (IFN) production. The cGAS enzyme acts as a unique sensor for double-stranded DNA. When DNA binds to cGAS it activates the enzyme producing a dinucleotide cGAMP from ATP and GTP. cGAMP created by cGAS activates STING (stimulator of interferon genes), leading to IFN immune response in the cell.
Transcreener CD73 Activity Assay
The Transcreener CD73 Assay measures adenosine (ADO) produced by ecto-5′-nucleotidase (also known as 5′-nucleotidase, NT5E, Cluster of Differentiation 73 or CD73). By measuring the production of ADO researchers can effectively determine the activity of the enzyme. The assay provides a powerful tool to screen entire compound libraries for CD73 modulators to help find new therapeutics for diseases such as cancer.
1600 Compound Pilot Screen with CD73
Acquire Accurate Dose-Response Measurements
Transcreener CD39 Activity Assay
The Transcreener AMP² CD39 Assay directly measures AMP produced the enzyme. By measuring the production of AMP researchers can effectively determine CD39 activity. Ectonucleotidases are plasma membrane-bound enzymes with externally-oriented active sites that metabolize nucleotides to nucleosides and are crucial for maintaining immune homeostasis. CD39 hydrolyzes ATP and ADP to AMP. AMP can further be processed to adenosine leading to a significant impact on many disease states. Recent studies have shown a key role for adenosine in immunosuppression in the tumor microenvironment, and ectonucleotidases are emerging as promising immuno-oncology targets.
Transcreener ENPP1 Activity Assay
ENPP1 is a nucleotide pyrophosphatase and phosphodiesterase that regulates purinergic signaling and can degrade different nucleotides including ATP and cGAMP. The Transcreener AMP²/GMP² ENPP1 Assay directly measures the GMP and (or) AMP produced by the enzyme. ENPP1 has been identified as the primary enzyme responsible for degrading cGAMP and therefore is under intense investigation as a therapeutic target for cancer immunotherapy.
cGAMP Titration with ENPP1
Human IKK-beta Titration When Converted to ADP Formed
Transcreener IKK-beta Activity Assay
IKK-beta is a serine kinase in the NF-κB pathway. IKK aids in the activation of NF-κB by phosphorylating a bound inhibitor IκBα, allowing NF-κB to enter the nucleus, in turn, activating various inflammatory responses. The signaling pathway is part of the innate immune response to bacterial and viral material, among other stimuli. Incorrect regulation can lead to a number of conditions including inflammatory autoimmune disorders and cancer.
Transcreener TBK1 Activity Assay
More than 30 known proteins are known to interact with TBK1 on various levels. One of the most studied features is the innate immune response to foreign infectious material. Toll-like receptors activate the enzyme leading to further signaling. As a regulator of the immune response pathway to viral and bacterial material, TBK1 interacts with TANK and TRAF3. TBK1 then phosphorylates interferon regulator factors (IRFs), which activate inflammatory immune response with type I IFNs. Its role in inflammatory modulation makes TBK1 a critical target for autoimmune and neurodegenerative diseases. TBK1 also regulates aspects of cell proliferation and death, making it a key focus area in cancer research.
Example Dose-Response with the Inhibitor Staurosporine
Robust Z’ Values
Transcreener JAK3 Activity Assay
JAK3 is a tyrosine kinase that is activated by cytokine receptors. JAK3 phosphorylates and activates STATs that induce the expression of genes in the cell nucleus. JAK3 can also autophosphorylate in the presence of ATP. Because it is expressed only in hematopoietic cells, which should help to limit off-target effects, JAK3 is also being vetted for autoimmune diseases like rheumatoid arthritis. A JAK1 and JAK3 inhibitor, Tofacitinib is currently marketed by Pfizer. New specific JAK3 inhibitors could help limit the current off-target effects of the drug.
Enzyme Titration with the Transcreener OAS1 Assay
As a first line of defense in the innate immune system, OAS1 is activated by viral double-stranded RNA. Upon activation, OAS1 synthesizes 2-5A from ATP which in turn activates RNase L. The result of this signaling event cleaves both viral and host RNA and in turn inhibition of protein synthesis in the cell, thus ending the replication of invading RNA viruses. The assay uses a coupling enzyme which generates AMP from the 2-5A. The AMP is then measured by the proven Transcreener AMP/GMP Assay with an FP readout.
Enzyme Titration with the Transcreener OAS1 Assay
Assay Development and Hit-to-Lead Services
JAK3 is a tyrosine kinase that is activated by cytokine receptors. JAK3 phosphorylates and activates STATs that induce the expression of genes in the cell nucleus. JAK3 can also autophosphorylate in the presence of ATP. Because it is expressed only in hematopoietic cells, which should help to limit off-target effects, JAK3 is also being vetted for autoimmune diseases like rheumatoid arthritis. A JAK1 and JAK3 inhibitor, Tofacitinib is currently marketed by Pfizer. New specific JAK3 inhibitors could help limit the current off-target effects of the drug.
Lead Discovery Services Include:
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