Monoclonal antibody purification

Monoclonal antibodies (mAbs) represent the largest and most progressive biopharmaceutical market. The mAbs used for clinical applications are subject to very high purity requirements, and there is, thus, a demand for more efficient and economical purification processes. Since mAbs are mainly used in therapy, their purity requirements are, therefore, extremely stringent. Today, the most widely used purification technique is affinity chromatography, and protein A resins are the products of choice for antibody capture. Protein A affinity chromatography techniques generate both high yield and high purity of antibodies in a single step. 

WorkBeads affimAb is an optimized alkali-stable protein A resin designed for the purification of monoclonal- and polyclonal antibodies in laboratory to process scale development. This resin has a superior base matrix in combination with an optimized alkali-stable protein A ligand. This results in high dynamic binding capacity also at short residence times, and stabile capacity over multiple purification cycles with cleaning-in-place using 0.5 M NaOH. Compared to the market-leading resin, this resin allows for higher purity of eluted mAbs from cell supernatants.

Purification of antibodies from mammalian host cells, such as Chinese Hamster Ovary (CHO), results in extensive bioburden on the protein A column. Chromatins, together with host cell proteins in general, cause damage to the protein A column. Regular cleaning-in-place (CIP) is mandatory in the purification process, but accumulative fouling of the column will still occur. The maximum lifetime of the protein A column is thus an important requirement. It would be desirable to add a purification step before the protein A purification step to reduce the components that constitute the worst threat and, in the process, improve the final purity of the target antibody. 

WorkBeads 40 TREN is a multimodal ion exchange chromatography (IEX) resin with a ligand (tris(2-aminoethyl)amine, TAEA) that is positively charged below pH 9. The resin has been demonstrated to be effective in the removal of impurities such as chromatin complexes (nucleic acid-protein complexes), endotoxins, and host cell proteins. We introduced WorkBeads 40 TREN upstream of WorkBeads affimAb in the flow scheme and used it in flow through mode to adsorb undesirable impurities while letting mAb pass through the column without binding.