Creative Biolabs is a leading preclinical biopharmaceutical company offering high-quality services and comprehensive solutions for target identification of small molecules. We have achieved unparalleled success in identifying target molecules by our unique target deconvolution technology and won a good global reputation. We will continue to build our global reputation and the team is rapidly expanding to keep pace with the growing demand for its services.
Radioactive Compound-Binding Assay
Molecular target identification of small molecules, also called target deconvolution, is a major step in phenotype-based drug discovery. For target deconvolution, biochemical purification based on radioactive (such as ³H-labeled) compound-binding assay with compound immobilized beads has been used for a long time (Fig.1). In this method, the compound is derivatized to have a reactive group, typically an NH2- or SH- group, and the derivatized compound is immobilized on the beads. A protein mixture (such as cell lysate) is placed on the compound-immobilized beads to enrich the proteins bound to the compound, and then the enriched protein is identified by mass spectrometry (MS)-based proteomics.
Fig.1 Affinity purification using compound-immobilized beads. (Kubota, 2019)
There are a variety of methods to identify drug targets and unintended off-target effects to understand the action mechanism of drugs. Target deconvolution based on radioactive compound-binding assay and its combination with other methods offers a novel methodology for the identification of small molecule targets, the discovery of drug-responsive biomarkers, and disease pathway analysis for preclinical target validation. Creative Biolabs is an expert in target deconvolution, and we provide highly specific radioactive compound-binding assays and their integration with other target deconvolution methods. These target deconvolution methods can be used for the identification and validation of novel drug targets.
![Binding assay of [³H]- compound 2 with recombinant proteins.](images/Radioactive-Compound-Binding-Assay-2.png)
Fig.2 Binding assay of [³H]- compound 2 with recombinant proteins. (Sasaki, 2018)
A good example is that Masato et al. reported the successful identification of pharmacological molecular targets for hepcidin production inhibitors by affinity purification and a radioactive compound binding assay. In this study, recombinant protein was incubated with [³H]- compound 2 after 2 h of preincubation with the unlabeled compound 2 as a competitor. The protein-binding radioactive compound was separated by a gel filtration column and measured by scintillation counting as disintegrations per minute (DPM). The experiment was performed twice independently and Fig.2 shows the expected results.
Key-benefits for radioactive compound-binding assay
Creative Biolabs’ scientists have excellent experience in target deconvolution, proteomics, computational biology, and drug discovery, and they can provide fast deliverables with analyzed results to save your time. In addition, Creative Biolabs supports your drug and biomarker discovery projects with end-to-end solutions tailored to the specific needs of your project. For more information, please feel free to contact us.
References
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