I have been screening the activity of immobilized enzymes in shaken tubes and need to move into testing activity under flow in a column. I am looking for recommendations on what column / pump / heat source I can use. I need to:
1) Keep the feed solution and columns warm at 50-60C
2) Use 50-100 mL of resin inside the column
3) Keep the flow through the column consistent at ~0.5 mL/min for at least 20 days
I tried using a Pharmacia peristaltic pump with 3 mm flexible tubing hooked up to a 100 mL Econocolumn. This was all inside an incubator to keep it warm. The trouble is that at such a low flow rate small fluctuations in the height of the liquid in the column or the degree of packing (it settles more tightly over time) cause the back pressure and flow rate to change and so the enzymes do not have a consistent supply of substrate.
Has anyone set up a really dependable column with low flow like this?
Did I mention is should be affordable? I would like to set up 10 of them!
Thanks!
-Jonathan
Dear Jonathan,
if I understand your problem right, your problem is the changing back pressure. Could a change in bead size solve your problem?
For instance our company is offering so-called XL particles (diameter 300 - 500 µm) with very low back pressure. The particles are available with standard functionalization (Ni-NTA, Glutathione, ...), but can also delivered custom-functionalized, like NHS activated and the like.
The link is
https://cube-biotech.com/products/purification-resins/xl-agaroses
Best, Roland
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