The Superdex S75 10/300 column is exhibiting abnormally high backpressure, with the system pressure rising to ~160 psi while maintaining a very low flow rate of only 0.040 mL/min. Despite multiple cleaning attempts using standard procedures (high-salt washes, low/high pH buffers, ethanol, and NaOH treatments), the issue persists and normal flow cannot be restored.
Any suggestions on how to clean and restore the performance?
In my experience, by the time you get to the described conditions, it's too late. You can try pumping in protease solutions (trypsin, papain, etc.) stopping the flow overnight to let digestion occur, then slowly flush. The problem is protein aggregates are going to be highly resistant to digestion. It also may not be only protein but lipids also in which case you need lipases too.
Dear Abhilek Kumar Nautiyal
did you try to invert the flow and wash the coloumn in the opposite direction?
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