Monoclonal antibodies (mAbs) are widely used as biopharmaceuticals. MAbs are easy to undergo structural and chemical changes during preparation and storage processes, and such denaturation may cause loss of therapeutic efficacy or manifestations of toxicity.  Therefore, therapeutic mAbs must be subject to strict quality control.

Size exclusion chromatography (SEC) is a powerful and convenient tool for determining mAb impurities in terms of molecular mass including aggregates, oligomers and fragments. We have developed three silica-based SEC columns designed especially for mAb analysis:

1) A 4.6 mm ID x 15 cm L semi-micro column packed with 25 nm pore size, 4 um particles, which is designed for high-throughput analysis of mAbs. The column realized high-throughput separation of mAb monomer and dimer in 3 min with complete separation, Rs>1.9.

2) A 7.8 mm ID x 30 cm L analytical column packed with the same particles as mentioned above. The column is suitable for high-resolution analysis of mAb monomer and dimer. In addition to complete separation of mAb monomer and dimer, mAb fragment with c.a. 100kDa could be partially separated from the monomer on this column.

3) A 7.8 mm ID x 30 cm L analytical column packed with newly developed 30 nm pore size, 3 um particles. Larger pore size with the exclusion limit of 2.5x10^6 Da provides improved separation and quantitation of mAb aggregates. Possible mAb pentamer could be partially separated on this column at lower flow rate of 0.2 mL/min.

We report here the features of these new SEC columns and their superior performance of mAb separation in comparison to conventional columns.