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peak purity in SEC, IEC, and Affinity chromatograph
Posted: Sun Jan 30, 2011 1:32 am
by pph31
How do you ensure the peak purity in the biologics seperation? such as peak purity in SEC, IEC, and Affinity chromatograph.
Will UV and Mass purity be helpful or doable considering the complexity of the biologics (mononer, dimer, aggregates, charged isomer)?
Thanks
Nick
Re: peak purity in SEC, IEC, and Affinity chromatograph
Posted: Sun Jan 30, 2011 3:16 am
by tom jupille
Assuming you're lalking about proteins, UV is not very helpful, because you're looking at the same UV absorbing amino acids; there are some variations, but nowhere near what you would need in order to be definitive.
MS (or, more properly MS/MS) on the other hand, would be definitive. And, yes, it's doable (but not easy). There is a great deal of work going on these days in LC-MS/MS for proteomics.
Re: peak purity in SEC, IEC, and Affinity chromatograph
Posted: Mon Jan 31, 2011 1:55 pm
by pph31
Thanks.
Can anyone from biologics separation suggest how you would do the peak purity check? or you never did because it is not required, or not critical in biologics separation?
Again, I appreciate your input, it bothers me.
Assuming you're lalking about proteins, UV is not very helpful, because you're looking at the same UV absorbing amino acids; there are some variations, but nowhere near what you would need in order to be definitive.
MS (or, more properly MS/MS) on the other hand, would be definitive. And, yes, it's doable (but not easy). There is a great deal of work going on these days in LC-MS/MS for proteomics.