Creative Biolabs has been a long-term expert in the field of antibacterial and antifungal drug discovery. Based on our extensive experience and world-leading technology platforms, Creative Biolabs is always dedicated to assisting our clients with the most satisfactory macromolecular synthesis assay for MOA study.
Mechanism of action (MOA) determination has been the most important component for antibacterial and antifungal drug discovery and development. In general, if a candidate compound is structurally similar to a known class of antibiotics, its target and MOA can be easily predicted. Subsequently, it can be verified by using a macromolecular synthesis assay.
Most antimicrobial drugs currently in clinical use selectively target and block one or multiple macromolecular synthesis sites, such as fatty acid (lipid) biosynthesis, protein synthesis (translation), DNA replication, RNA synthesis (transcription), and cell wall (peptidoglycan) synthesis. In addition, there are also a few drugs that work by interfering with other mechanisms such as ion channels and bacteriolysis.
Fig.1 Sites of antibacterial action and mechanisms of resistance. (Brown, 2015)
To perform this assay, radiolabeled precursors accumulated by the test strain cells would be incorporated into specific macromolecules. Since free precursors are soluble in trichloroacetic acid (TCA) and synthetic macromolecules are not, the radiolabeled macromolecules can be selectively precipitated and isolated, and then quantified by scintillation counting to gain insight into which pathways are inhibited.
During this assay, antibiotics known to disrupt a particular pathway can be used as both positive controls for compounds testing that pathway and negative controls for other pathways. For example, irgasan and cerulenin can be used as the positive controls for fatty acid biosynthesis pathway, tetracycline and linezolid can be used as the positive controls for protein synthesis pathway, ciprofloxacin can be used as the positive control for DNA replication, rifampin can be used as the positive control for RNA synthesis, imipenem and vancomycin can be used as the positive controls for cell wall synthesis. In recent years, we have performed multiple validations in E. coli and Staphylococcus aureus and have gradually expanded to more strains.
Equipped with world-leading technology platforms and professional scientific staff, we are pleased to offer the most qualified services and products for antibacterial and antifungal drug discovery. If you are interested in our products or services, please do not hesitate to contact us for more detailed information.
Reference
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