Abstract
Hydroxyapatite chromatography is shown to be an excellent method for chromatographically purifying monoclonal antibodies (Mab). Mab contained in eluates from Protein A columns was partially purified on ceramic hydroxyapatite (CHT™) Type I, 40 μm ceramic hydroxyapatite using two scouting methods which provide milligram amounts of Mab typical at laboratory scale. The result from one of the scouting methods was optimized to obtain a high concentration of purified Mab with acceptable clearance of cell culture impurities. Several techniques (linear phosphate screening, linear alkaline salt screening, and two alkaline salt step gradients) are described for obtaining high concentrations of purified Mab in a lab-scale CHT chromatography column.
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References
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Cummings, L.J., Frost, R.G., Snyder, M.A. (2014). Monoclonal Antibody Purification by Ceramic Hydroxyapatite Chromatography. 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_15
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DOI: https://doi.org/10.1007/978-1-62703-992-5_15
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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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