The ultrafiltration/diafiltration is the last step for protein purification. During the ultrafiltration/diafiltration of protein solution, i found TWEEN 80 was adsorbed by membrane. I want to add the TWEEN 80 after diafiltration. Can somebody give me some advice?
Start with a 10% stock solution of Tween-80 in water, rather than the pure detergent, because the viscosity of the pure detergent makes it difficult to dispense and mix. I recommend you use the Surfact-Amps-80 product, which is sealed in ampoules to prevent the accumulation of protein-damaging oxidation products.
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If you saturate your diafiltration membrane with Tween-80 before you apply your sample, you shouldn't loose any detergent during the diafiltration step.
You could try to run a batch of sterile sample buffer (with Tween-80) through the filtration cell. Once the membrane is saturated with Tween-80, the detergent concentration in your sample should remain more or less constant.
Sounds like the pore size is too small for the Tween-80 micelles. Can you use a larger pore size? Alternatively, would it be possible to use a different detergent with a smaller aggregation number (= micelle size)?
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