Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.chroma.2009.09.059
Title: Engineering of a two-step purification strategy for a panel of monoclonal immunoglobulin M directed against undifferentiated human embryonic stem cells
Authors: Tscheliessnig, A.
Ong, D.
Lee, J.
Pan, S.
Satianegara, G.
Schriebl, K.
Choo, A. 
Jungbauer, A.
Keywords: Adsorption isotherm
Adsorption kinetics
Anion-exchange chromatography
Benzonase
IgM
Number of binding sites
PEG
Precipitation
Issue Date: 6-Nov-2009
Source: Tscheliessnig, A., Ong, D., Lee, J., Pan, S., Satianegara, G., Schriebl, K., Choo, A., Jungbauer, A. (2009-11-06). Engineering of a two-step purification strategy for a panel of monoclonal immunoglobulin M directed against undifferentiated human embryonic stem cells. Journal of Chromatography A 1216 (45) : 7851-7864. ScholarBank@NUS Repository. https://doi.org/10.1016/j.chroma.2009.09.059
Abstract: A two-step purification strategy comprising of polyethylene glycol (PEG) precipitation and anion-exchange chromatography was developed for a panel of monoclonal immunoglobulin M (IgM) (pI 5.5-7.7) produced from hybridoma cultures. PEG precipitation was optimized with regards to concentration, pH and mixing. For anion-exchange chromatography, different resins were screened of which Fractogel EMD, a polymer grafted porous resin had the highest capacity. Despite its significantly slower mass transfer, the binding capacity was still higher compared to a convection driven resin (monolith). This purification strategy was successfully demonstrated for all 9 IgMs in the panel. In small scale most antibodies could be purified to >95% purity with the exception of two which gave a lower final purity (46% and 85%). The yield was dependent on the different antibodies ranging from 28% to 84%. Further improvement of recovery and purity was obtained by the digestion of DNA present in the hybridoma supernatant using an endonuclease, benzonase. So far this strategy has been applied for the purification of up to 2 l hybridoma supernatants. © 2009 Elsevier B.V. All rights reserved.
Source Title: Journal of Chromatography A
URI: http://scholarbank.nus.edu.sg/handle/10635/67035
ISSN: 00219673
DOI: 10.1016/j.chroma.2009.09.059
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