The strong acidic elution conditions needed for Protein A affinity chromatography, commonly used for the capture of mAbs from clarified tissue culture fluid, can trigger structural changes and promote the oligomerization of pH-sensitive molecules. Elevated levels of antibody aggregates are often associated with the overproduction of mAbs due to the mispairing of disulfide bonds and the unfolding or denaturation of drug molecules at cell growth temperatures (25☌ or above) ( Rathore et al. This has had a significant impact on downstream processing. However, these advancements have often adversely affected impurity composition and concentration upon harvest. Over the years, upstream technology advancements have helped improve the titer of target antibodies, from merely 1 g/L two decades ago to 10–13 g/L in fed-batch processes and up to 25 g/L in perfusion cultures today ( Gronemeyer et al. Monoclonal antibodies (mAbs) remain a predominant class of therapeutic protein products on the market because of their wide range of applications in disease treatment and diagnosis. Comparison of CHT, Capto adhere, and Capto adhere ImpRes in the clearance of mAb S aggregates Pooled elution fractions from the Capto adhere ImpRes Column with 50 mM sodium acetate buffer at different pH.Ĭlearance of Aggregates by the Different Mixed-Mode Media Monomer Content and Recovery from Capto adhere Impres Media under Various pH Conditions Pooled elution fractions from the Capto adhere Column using 50 mM sodium acetate buffer at different pH. Monomer Content and Recovery from Capto adhere Media under Various pH Conditions Elution of mAb S monomers from a Capto adhere ImpRes Column Monomer Elution from Capto adhere ImpRes Mediaįigure 6. Purification of mAb S on a Capto adhere ImpRes Column Monomer Purification Using Capto adhere ImpRes Mediaįigure 5. Elution of mAb S monomers from a Capto adhere Column Purification of mAb S on a Capto adhere Columnįigure 4. Monomer Purification Using Capto adhere Mediaįigure 3. Elution of mAb S monomers from a CHT Column Purification of mAb S on a CHT Columnįigure 2.
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