Hello,
If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange?
For example, if i flow through 500 uL of 1mg/mL sample, and collect the 700 uL of buffer elution, will the concentration of the elution always be less than 1 mg/mL? or will there be time when an antibody recovery after buffer exchange is 100% e.g. 1mg/mL elution
There is usually some loss of protein. This can be reduced by collecting more of the tail of the protein peak, but potentially at the cost of less efficient buffer exchange.
Two other methods of buffer exchange are dialysis and diafiltration (concentrating by ultrafiltration, rediluting, and concentrating again). The yield is better than gel filtration, but they take a lot longer.
Yes I aggree with the Answer of Adam B.Shapiro , during th Elution pattern the elute volume size should decrease as well as collect upto the reaching of base lines.
And depends on the Elution buffer strength, Diafiltration using with the relevent Nominal Molecular Weight Limit (NMWL)Cassete system by the priniciple of Tangential Flow Filtration (TFF) with approprite Low protein Binding membrane
Hi there,
Size Exclusion Chromatography, which is the main application of Sephadex, has always a diluting effect due to the diffusion of the sample molecules during the process.
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