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Development of a multi-product leached protein A assay for bioprocess samples containing recombinant human monoclonal antibodies.
Development of a multi-product leached protein A assay for bioprocess samples containing recombinant human monoclonal antibodies.
Ren, D., Darlucio, M.R., Chou, J.H.
Journal
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J Immunol Methods
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Species
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Analytes Measured
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NT-proANP
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Matrix Tested
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Assay diluent
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Year
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2011
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Volume
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366
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Page Numbers
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20-7
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Application
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Abstract
The detection of low level of protein A leached from monoclonal antibody downstream purification process is often interfered by the presence of excess amount of product antibody. In order to prevent this interference, we developed a new multi-product leached protein A assay that used acidification to completely dissociate the IgG-ProteinA complex, followed by neutralization under selected condition to prevent re-formation of the IgG-ProteinA complex. The amount of protein A was then determined by a sandwich immunoassay using Meso Scale Discovery technology. The assay takes approximately 3h to complete for one 96-well plate of samples, and this has been successfully applied to samples containing different monoclonal antibody products examined so far. The data demonstrates that this assay is robust and efficient in determining leached protein A contamination during purification of recombinant monoclonal antibodies.
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