MEP Antibacterial Screening Service
The methylerythritol phosphate (MEP) pathway, is utilized by most bacteria, including all Gram-negatives and many Gram-positives, and plant chloroplasts while the mevalonate pathway is found in humans, plant cytosol and some Gram-positive bacteria. Due to this natural distribution, the MEP pathway represents a promising target for development of novel antibacterial agents and herbicides.
- Echelon has 25 years of assay development expertise and 15 years experience in the MEP field.
- Your samples are cataloged and stored according to your requirements.
- The MEP Screening Service consists of two screens; a primary screen to identify “hits” of greater than 50% inhibition and a secondary screen to test any inhibitor identified in the first screen for specificity towards the MEP pathway.
- Usually, results are provided 2 weeks from receipt of samples. We can accommodate tighter schedules if needed.
Contact us for a quote on your custom assay service.
Description
The MEP Screening Service consists of two parts:
- The “Primary Antibacterial Screen” identifies initial “hits”, >50% inhibition, at an initial concentration (usually 100 µg/mL). We screen up to 80 compounds per screen in duplicate.
- The “Secondary MEP-Selectivity Screen”: A potential inhibitor, identified above, is further tested against our engineered cell line under two growth conditions to determine if the potential inhibitor displays specificity toward the MEP pathway. Dose-response experiments are performed on 4 compounds per plate in duplicate and a MIC value (minimum inhibitory concentration), >90% inhibition, is assigned to each potential inhibitor.
Recommended concentration of compound provided by customer is 5-10 mg/mL in DMSO. DMSO is well tolerated with no effect at up to a 2% final concentration for both MEP and MVA pathway growth. The assay has been tested for the measurement of MIC values for known MEP pathway inhibitors. Known antibiotics not targeting the MEP pathway exhibit MICs comparable to wild-type S. typhimurium.
The compound should be provided in a DMSO solution in a glass vial or libraries in 96-well plates indicating the concentration in mg/mL. The concentration has to be at least 25 times higher than the highest concentration to be tested. A sample volume of 200 µL will be sufficient for both tests. Samples in DMSO should be in solid form at 4 ºC, otherwise there is moisture in the DMSO and the compound concentration may be inaccurate. Please ship samples on blue ice or dry ice. Upon receipt, all samples will be stored at -20 ºC
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Publications
Publications for this service are based off of the MEP Synthase Inhibitor Screen (catalog number K-2000C), the product used for this assay service.
1) Bochar, D.A.; Freisen, J.A.; Stauffacher, C.V. and Rodwell, V.W.(1999) Comprehensive Natural Products Chemistry,(Cane, D. Ed.) Pergamon Press, Oxford, pp. 15-44.
2) Sacchettini, J.C. and Poulter, C.C. (1997) Science, 277(5333), 1788-9.
3) Testa, C.A.; Brown, M.J. (2003) Current Pharmceutical Biotechnology, 4, 248-259.
4) Koppisch, A.T.; Fox, D.T.; Blagg, B.; Poulter, C.D. (2002), Jan 8; 41 (1), 236-43.