The antibody therapeutics market is enjoying high growth rates, the major areas of therapeutic application being cancer and immune/inflammation-related disorders. Six of the top ten best-selling global drug brands are monoclonal antibody-based. This market is predicated to show continued growth for many years to come, with more monoclonal antibodies (mAbs) designed and produced for treatments of specific diseases.

Early in mAb development there are many harvested cell supernatant samples which contain mAbs that are secreted by cell cultures. These samples must be screened for mAb titer; affinity protein A columns are often employed for this purpose. Determination of the optimal time for harvesting mAbs from cell culture supernatant and screening for the best cell clones that express the most amount of mAb are also accomplished using protein A columns. In addition, partial purification of mAb can be accomplished using an affinity protein A column initially to establish the right cell lines and to partially characterize a newly produced mAb. With many samples to be screened for different purposes, a reliable and high throughput column is needed for this workflow.

In this application note, the quick capture and accurate titer analysis over a wide concentration range of mAb is demonstrated using a TSKgel Protein A-5PW, 20 μm analytical column. Packed with hydroxylated methacrylic polymer beads coupled with a recombinant Protein A ligand (a code-modified hexamer of the C domain), this 4.6 mm ID × 3.5 cm PEEK column can be used with high flow rates for high throughput analysis and still maintains chromatographic efficiency, peak width and resolution. In addition, the TSKgel Protein A-5PW column can perform for more than 2,000 injections with no sign of deterioration and without cleaning.