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- Posts: 2
- Joined: Thu Jun 01, 2023 8:34 am
- Location: Scotland
I am running SEC to check for antibody purity on an Agilent 1260 and am using a a Biosep SEC s3000 column for the run. The absorbance for the run is a280 at 1ml/min. Bandwith is set at 8.
The mobile phase for the run is 50mM NaPO4 NaN3.
I work in a manufacturing laboratory (not research). To meet our in house specifications the Monoclonal Antibodies must have retention times that fall between set positions according to the Bio-Rad Standards. In this case Thyroglobulin (MW670,000) and gamma-globulin (MW158,000). Generally the first 2 of the 5 peaks you expect to get when using this standard.
The issue I am having is that the standards are appearing unresolved and not separated on my traces (See Attached Link). In simple terms a big blob with some peaks at the top. You will also see an image of a 2mg/ml BSA standard I ran for reference.
If anyone has any troubleshooting tips for this issue it would be a huge help to me. I have plenty experience running the instrument, but little in terms of method creation or research.
https://imgur.com/a/zsU4eTZ