Calibration Curves for TSKgel® SW mAb Columns
Materials
analytical column
Application
guard column
mobile phase component
standard
CONDITIONS
column
TSKgel SuperSW mAb HR, 30 cm x 7.8 mm I.D., 4 μm particles (822854)
column
TSKgel UltraSW Aggregate, 30 cm x 7.8 mm I.D., 3 μm particles (822856)
column
TSKgel G3000SWxl, 30 cm x 7.8 mm I.D., 5 μm particles (808541)
mobile phase
0.2 M sodium phosphate buffer, pH 6.7 and 0.05% sodium azide
flow rate
1.0 mL/min
column temp.
Ambient
detector
UV, 280 nm
injection
10 μL
Description
Analysis Note
In gel fitration chromatography, the calibration curve of a column shows the relationship between retention time or retention volume of a set of water-soluble polymer or protein standards vs. the logarithm of their molecular masses. The linear portion of a calibration curve is used to estimate the molecular mass of an unknown protein. An expanded linear range is desirable as it allows for a wider variety of molecular masses to be analyzed with that column. This example shows the calibratuion curves for TSKgel® SuperSW mAb HTP, TSKgel SuperSW mAb HR, TSKgel UltraSW Aggregate and the industry standard TSKgel G3000SWxl column using several commonly available globular proteins and p-aminobenzoic acid as a small MW marker to indicate the void volume of the column. The exclusion volume of the column is indicated by a trimer of thyroglobulin. [Note that the calibration curve of the 15 cm x 4.6 mm ID TSKgel SuperSW mAb HTP column is not included in the Figure as the column volume is every small and the packing material in the column is the same as the TSKgel SuperSW mAb HR column].
Legal Information
TSKgel is a registered trademark of Tosoh Corporation