Does Waters have a calibration curve for ACQUITY Premier Protein SEC 250A and XBridge Premier Protein SEC 250A columns? - WKB227536
ENVIRONMENT
- Size exclusion chromatography (SEC)
- Calibration curves
- ACQUITY Premier Protein
- XBridge Premier Protein
ANSWER
No
ADDITIONAL INFORMATION
Waters does not have calibration curve data for our new ACQUITY Premier Protein SEC 250A or XBridge Premier Protein SEC 250A columns.
Waters finds that calibration curves for SEC columns may not provide a full picture of the capabilities of a particular packing.
For example, the calibration curve for BEH450 SEC columns does NOT indicate that they would work for the SEC separation of AAVs.
The BEH SEC calibration curve can be seen in these documents:
ACQUITY UPLC BEH SEC Columns and Standards Care & Use Manual
XBridge Protein BEH SEC Columns & Standards Care & Use Manual
If you look at the following document, you will see that even though AAVs are well above the MW range of the BEH450 SEC columns, they are able to be separated with a BEH450 SEC column:
Size-Exclusion Chromatography Analysis of Adeno-Associated Virus (AAV) Preparations Using a 450 Å Diol-Bonded BEH Column and Fluorescence Detection
In summary:
Calibration curves for biomolecule separations on SEC columns, based on pore diameter, are not the only indicator of how successful an SEC method might be for a given MW biomolecule.
Choice of pore size should be based empirically for a given analyte.
Also see:
ACQUITY Premier Protein SEC Columns Care & Use Manual
XBridge Premier Protein SEC Columns Care & Use Manual
Application Notes:
Reliable High Resolution Protein SEC Separations for Online Native LC-MS mAb Analysis
USP Monograph mAb SEC Method Robustness on the XBridge Premier Protein SEC Column
id227536, monoclonal antibody