Abstract
Magnetic microspheres represent an interesting alternative to conventional chromatography resins in automated high-throughput protocols replacing centrifugation and filtration by magnetic separation. Some magnetic microspheres have unique features like high magnetite content and non-porous surface which allows them to migrate very fast in magnetic fields while binding target molecules with a low unspecific adsorption. Here, we describe the use of protein A or protein G-coated magnetic microspheres to purify quickly monoclonal antibodies from crude serum-free supernatants without the need of preliminary clarification or purification step. Using this method, multiple samples can be processed in parallel, a high level of purity can be obtained, and the purified IgG maintain their biological activity.
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References
Holschuh K, Schwämmle A (2005) Preparative purification of antibodies with protein A—an alternative to conventional chromatography. J Magn Magn Mater 293:345–348
Gao J, Li Z, Russell T, Li Z (2012) Antibody affinity purification using metallic nickel particles. J Chromatogr B 895–896:89–93
Hafeli UO (2003) Characterization of magnetic particles and microspheres and their magnetophoretic mobility using a digital microscopy method. Eur Cell Mater 3(2):24–27
Merck Estapor Microspheres (2011) A versatile biotool for bioseparation, biopurification and biodetection, Bio-Estapor® microspheres, Brochure
Carriere D (1999) Whole blood Capcellia CD4/CD8 immunoassay for enumeration of CD4+ and CD8+ peripheral T lymphocytes. Clin Chem 45:92–97
Francois P (2003) Rapid detection of methicillin-resistant Staphylococcus aureus directly from sterile or nonsterile clinical samples by a new molecular assay. J Clin Microbiol 41(1):254–260
Stege PW, Raba J, Messina GA (2010) Online immunoaffinity assay-CE using magnetic nanobeads for the determination of anti-Helicobacter pylori IgG in human serum. Electrophoresis 31(20):3475–3481
Safarik I, Safarikova M (2004) Magnetic techniques for the isolation and purification of proteins and peptides. Biomagn Res Technol 2:7
Pande J, Szewczyk MM, Grover AK (2010) Phage display: concept, innovations, applications and future. Biotechnol Adv 26(6):849–858
Josic D, Lim Y-P (2001) Methods for purification of antibodies: analytical and preparative methods for purification for antibodies. Food Technol Biotechnol 39(3):215–226
Acknowledgments
This work was supported by Fabrice Sultan from Merck Estapor® who provided us with all magnetic microspheres.
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© 2014 Springer Science+Business Media, New York
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Malinge, P., Magistrelli, G. (2014). Rapid Purification of Monoclonal Antibodies Using Magnetic Microspheres. In: Ossipow, V., Fischer, N. (eds) Monoclonal Antibodies. Methods in Molecular Biology, vol 1131. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-992-5_16
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DOI: https://doi.org/10.1007/978-1-62703-992-5_16
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-991-8
Online ISBN: 978-1-62703-992-5
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