We are producing recombinant proteins in E.coli and we are interested in resuspension of 50-1000 gr pellets in an easier way than using a vortex or pippette so we look for homogenizators or other alternatives. We are also interested in equipments for sonication of 1-10L volum. Do you have any recommendations?
Thanks!
Dear Juan
i think that the sonication is preferable when you have not so big biomass (eg.10-20g). To achieve an efficient lysis you need to re-suspend the pellet in at least 10ml/g and if you have 100g, it means that you have to sonicate 1liter of solution. This is not impossible but is quite difficult perform an homogeneous sonication and cool down the sample homogeneously to prevent protein unfolding due to temperature increase. I think that when you have to manage so larger biomasses the FRENCH press cell is preferable.
best regards
Manuele
Juan Martinez-Olivan
Dear Juan
for my research thesis, I have used sonication with 8M urea, and if you decrease urea concentration, the clearness of the cell suspension will be less.
as Manuele said, I also recommend sonication with 8M urea if you using the denaturing method.
For such large amounts of material, the best method to use for cell breakage may be a microfluidizer. A French press limits you to a volume of about 30 ml, but a microfluidizer is a continuous flow instrument , so it can accommodate any volume of cell suspension.
I'm not sure what to use for resuspending huge amounts of cell pellet. Maybe a blender or food processor.
Ultrasonic cell disruptors deliver reliable and reproducible results for the lysis of E. coli. Ultrasonic probe-type homogenizers operate with approx. 20,000 cycles per second .Advantage of Ultrasonic Lysis for very small to very large samples (µL to litres).
Following on the comment by Rithab Al-Samawi , I looked for continuous-flow ultrasonic cell disruption equipment. Here is one:
https://www.sonicator.com/products/high-volume-continuous-flocell-for-q700-and-q1375
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