Jump to Main Content
Affinity chromatographic purification of human immunoglobulin M from human B lymphocyte cell culture supernatant
- Liu, Zhuo, Gurgel, Patrick V., Carbonell, Ruben G.
- Biochemical engineering journal 2013 v.70 pp. 63-70
- B-lymphocytes, antibodies, binding capacity, binding proteins, cell culture, chromatography, feedstocks, humans, immunoglobulin G, immunoglobulin M, ligands, molecular weight
- Compared to immunoglobulin G purification with extensively studied affinity ligands such as protein A and protein G, little work has been done on affinity chromatographic purification of immunoglobulin M. Hexamer peptide ligand HWRGWV, previously shown to bind specifically to the Fc fragment of IgG, also demonstrated potential for IgM purification. This study presents further characterization and investigation of this ligand for its potential for purification of IgM. Different running conditions were employed in order to improve the recovery and purity of IgM. The final recovery and purity of the antibody is feedstock dependent, but can reach levels of both recovery and purity as high as 95%. The dependence of the recovery and purity on total loading amount and initial IgM concentration were investigated and discussed. Although relatively low dynamic binding capacities (DBC) in the range of 4.6–13.1mg IgM/mL resin at linear flow rates from 173 to 35cm/h were obtained for IgM compared to IgG because of the large molecular weight of IgM, the DBC value of HWRGWV for IgM is much greater than protein-based IgM affinity ligands found in the literature and is competitive with current commercially available affinity ligands, such as KAPTIVE-M, CaptureSelect IgM and Ultralink Immobilized Mannan Binding Protein.