Large-scale preparation of purified monoclonal antibody from cell culture supernatant: A case study with a monoclonal antibody to Zeta globin chain

Background: Nowadays, monoclonal antibodies have become important tools used in biomedical research, diagnosis and treatment of diseases. The mAbs are isolated from cell culture media, which usually contain different proteins in addition to antibodies. Obviously, antibody purification is becoming critical to guarantee its reliable application. Protein A and G resins are the most efficient and widely-used ligands in chromatographic methods. Nevertheless, some mAbs could not be purified by Protein A or G column. Objectives: In order to explore the alternative method for purification of an IgG1 mAb, four different types of affinity chromatography were studied and compared in term of efficiency. Materials and method: Chromatography using four commercial ligands, Protein A, Protein G, Protein L and engineered recombinant Protein A, was employed to purify the anti-zeta globin chain mAb clone PL3. The performance of the chromatography was compared in terms of yield, purity and biological activity of mAbs. Result: The mAb PL3 could only be purified by using engineered recombinant Protein A column. The biological activity and purity of the purified mAbs were checked by ELISA and SDS-PAGE, respectively. It was found that the obtained purified mAbs were high purity and retained their biological activity. Conclusion: In conclusion, engineered recombinant Protein A column is an alternative technique for large-scale purification of mAbs produced by unusual hybridoma clone that could not be purified by other columns.