Affinity chromatographic purification of human immunoglobulin M from human B lymphocyte cell culture supernatant |
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| Institution: | 1. Urology and Nephrology Research Center, Department of Urology, Shahid Labbafinejad Medical Center, Shahid Beheshti University of Medical Sciences, Tehran 198396-3113, Iran;2. Cellular and Molecular Research Center, Zahedan University of Medical Sciences, Zahedan 98167-43181, Iran;3. Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan 98167-43181, Iran;1. Coastal and Freshwater Group, Cawthron Institute, Private Bag 2, Nelson 7012, New Zealand;2. School of Biological Science, Victoria University of Wellington, PO Box 600, Wellington 6140, New Zealand;3. Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland 1142, New Zealand;4. Institute of Marine Science, University of Auckland, Auckland 1142, New Zealand;1. Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA;2. The RNA Interference and Non-Coding RNA Center, The University of Texas MD Anderson Cancer Center, Houston, TX, USA;3. Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, TX, US;4. UMF Carol Davila Bucharest, Romania and Fundeni Hospital, Bucharest, Romania;5. Leukemia Department, Santa Anna Hospital, University of Ferrara, Ferrara, Italy;6. Hacettepe University Faculty of Medicine, Ankara, Turkey;7. Hereditary Cancer Program, Catalan Institute of Oncology, IDIBELL, Hospitalet de Llobregat, Barcelona, Spain;8. Molecular Morphology Laboratory, AC Camargo Cancer Center, São Paulo 01508-010, Brazil;9. Department of Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA;10. Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA;11. Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX, USA;12. Department of Emergency Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA;13. Department of Endocrine Neoplasia and Hormonal Disorders, The University of Texas MD Anderson Cancer Center, Houston, TX, USA |
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| Abstract: | 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 4.6–13.1 mg IgM/mL resin at linear flow rates from 173 to 35 cm/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. |
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