| Determining the mechanism of action (MOA) of new agents is essential to the discovery process and is also a required component of the Investigational New Drug (IND) filing. Micromyx utilizes a variety of whole cell, molecular, and biochemical techniques to determine the MOA: |
| Macromolecular Synthesis Labeling Whole cell assays involving specific radiolabeling of DNA, RNA, protein, cell wall, and lipid pathways. Inhibition of each pathway is examined using concentrations of the test agent above and below the MIC value. Models established in Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Clostridium difficile. In vitro Transcription/Translation Assay Antimicrobial agents that are shown to inhibit protein synthesis in the macromolecular synthesis assay (shown above) are further validated using an E. coli extract and luciferase reporter system. An IC50 value is generated and compared to the MIC of the test agent. |
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| Cell Permeability Assay Effect of Polymyxin B on ATP Release From Staphylococcus aureus ATCC 29213 Antimicrobial agents that target the cell membrane and cause leakage of essential small molecules are assessed by measuring the leakage of ATP into the growth medium as the cells are dosed with concentrations of the test agent above and below the MIC value. Enzyme Assays Micromyx can evaluate the ability of test agents to inhibit enzymatic reactions in either crude extracts or in purified enzyme systems. Red blood cell lysis Using red blood cells of any available orgin (human, sheep, mouse, etc.), test agents are dosed at levels above and below the MIC value. In order to assess lysis, hemoglobin release is measured using a spectrophotometer. |
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Micromyx WMed Innovation Center 4717 Campus Drive Center Kalamazoo, MI 49008 |
Phone: 269-372-3758
Fax: 269-353-5567 |