I'm working on a protein that is potentially harmful when it may form aerosols. Whenever I manipulate the sample openly, I'm therefore working under a flow bench (S2-Lab). I'm also directly sending the sample from the flow bench into the sample loop of an FPLC by tubing plugs attached to the bench. The problem is that the distance from the bench to the FPLC is so long that the sample gets diluted on the way... (I checked that by applying a dye instead of the sample first). What can I do about that ? I thought about applying the sample between air bubbles to avoid dilution, but this would get the air on my gelfiltration column, which is not desirable... Has anyone yet found a solution for this dilemma?
If I were you I would put everything under the biological hood, this is safest also if you suffer leakage in the FPLC tubing or you can try a sealed vial and an autosampler.
best regards,
francesco
Its better to setup the the entire experiment (FPLC run) within a hood or some kind of safety cabinet.Make sure the column waste ,other eluents and entire run (buffers) etc are monitored before being collected or discarded.
There is also the possibility to use tubing of smaller diameter. The smaller the diameter, the less dilution you will get!
Please use the smaller dia of the tube. Never let the air bubbles in to your column. It will create cracks in the matrix and you will not get proper results. try to reduce the distance and use it in a hood
If it is really a hazard I would definetly contain the FPLC in a biohazard hood as suggested and dont forget the waste stream. But instead of tubing and a pump why not inject the sample into a superloop - you would need to avoid any needle sticks.
What kind instrument do you use? Is it GE? If yes, I suggest you have it in cooled room. New instruments Bioline offer benchtop cooling so you can have very short path length. Bioline offers higher pressures and has its own low pressure control, if the air reaches pump delivering sample. In this case pump stops and the air cannot reach head of the GPC column.
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