Mechanistic modeling of hydrophobic interaction chromatography for monoclonal antibody purification: process optimization in the quality by design paradigm
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Purification of monoclonal antibodies by hydrophobic interaction chromatography under no-salt conditions
Hydrophobic interaction chromatography (HIC) is commonly used as a polishing step in monoclonal antibody purification processes. HIC offers an orthogonal selectivity to ion exchange chromatography and can be an effective step for aggregate clearance and host cell protein reduction. HIC, however, suffers from the limitation of use of high concentrations of kosmotropic salts to achieve the desire...
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This paper describes a method for the plasmid DNA purification, which includes an ammonium sulphate precipitation, followed by hydrophobic interaction chromatography (HIC) using Phenyl Sepharose 6 Fast Flow (low sub). The use of HIC took advantage of the more hydrophobic character of single stranded nucleic acid impurities as compared with double-stranded plasmid DNA.
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was quantified by integration using EZLogicTM integration software (Bio-Rad). The clearance of host cell proteins (HCP) and double-stranded DNAs (dsDNA) was determined by a CHO-CM HCP ELISA kit (Cygnus Technologies) and Quant-iT dsDNA high-sensitivity assay kit (Life Technologies Corporation), respectively. mAb1 concentration was determined by UV absorption at 280 nm using a coefficient of 1.4 ...
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Matrix-bound monoclonal antibodies against urokinase have been used to purify this enzyme by affinity chromatography. In a single-step procedure, urokinase can be isolated from crude preparations with high yield and high purity, and without loss of enzymatic activity.
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ژورنال
عنوان ژورنال: Journal of Chemical Technology & Biotechnology
سال: 2018
ISSN: 0268-2575
DOI: 10.1002/jctb.5742