SuperSW mAb系列色谱柱从杂质中分离单抗单体（AN51）
Introduction
The analysis of monoclonal antibodies (mAb) is growing in importance in the field of biotherapeutics for the treatment of a variety of diseases. Quality control of therapeutic mAb is essential, as the introduction of species to the body other than the monomer may induce toxic side effects. Therefore, the pure antibody monomer must be very well resolved from its dimer and higher molar mass aggregates, as well as the antibody fragments. Size exclusion chromatography (SEC) is the best choice for determining mAb monomers and their impurities, including aggregates, oligomers, and mAb fragments.
Tosoh Bioscience has answered the call for dedicated SEC columns for the high resolution separation of mAb with the new silica-based 4 μm TSKgel SuperSW mAb HR column, for high resolution separation of the monomer and dimer, and the 3 μm TSKgel UltraSW Aggregate column for the separation and quantification of mAb aggregates and oligomers. This application note demonstrates the superb performance of these new columns for the analysis of monoclonal antibodies.
Experimental Conditions
Column: TSKgel SuperSW mAb HR, 4 μm, 7.8 mm ID × 30 cm
TSKgel G3000SWXL, 5 μm, 7.8 mm ID × 30 cm
Mobile phase: 200 mmol/L potassium phosphate buffer + 0.05% NaN3,pH 6.7
Flow rate: 1.0 mL/min
Detection: UV @ 280 nm
Temperature: 25 ºC
Injection vol.: 10 μL
Sample: 10 g/L IgG digested with papain for 0-24 hr
Column: TSKgel UltraSW Aggregate, 3 μm, 7.8 mm ID × 30 cm
Mobile phase: 100 mmol/L potassium phosphate buffer, 100 mmol/L sodium
sulfate, pH 6.7 + 0.05% NaN3
Flow rate: 1.0 mL/min
Detection: UV @ 280 nm
Temperature: 60 ºC
Injection vol.: 20 μL
Sample: BI-mAb-02 (4.6 mg/mL)