Screening/Optimization of loading conditions for Capto adhere.
- An example
Capto™ adhere is a multimodal anion exchange medium designed for intermediate purification and polishing of monoclonal antibodies after capture on Protein A medium. The medium is designed to be run in flow-through mode where the antibodies pass through the column while key contaminants such as dimers/aggregates (D/A), viruses, host cell proteins (HCP), and leached protein A are adsorbed.
Balancing product yield against product purity is the major consideration when optimizing a method. The results of a screening experiment for optimal loading conditions for a given antibody are described below. The variables studied were load, conductivity of the feed, and pH of the feed. The responses monitored were antibody yield and concentration of D/A, HCP, and protein A in the flowthrough pool. The screening was done by use of design of experiments (DoE), using MODDE 7.0 software (Umetrics). The outputs from the DoE software are mathematical models describing the effect of the different loading parameters on the responses. These mathematical models were then uploaded to the website. The website is thus interactive in such a way that the user can experiment with the loading parameters and see how different parameter settings will affect antibody yield and contaminant clearance.
Note! The loaded parameter settings are dependent on antibody/antibody feed. The models shown only apply to the studied antibody.
General trends
Although each antibody/antibody feed will require different parameter settings, experience with different antibodies has shown that some general trends can be expected (Figure 1). High load will favor yield, while low load will favor clearance of D/A, protein A, and HCP. Note that we have found that D/A clearance is less affected by conductivity than the clearance of protein A and HCP.
Further reading
Instructions:
Capto adhere
HiTrap Capto adhere
Application Notes:
Optimization of loading conditions on Capto adhere using design of experiments
Selective removal of aggregates with Capto adhere
Two-step purification of monoclonal IgG1 from CHO cell culture supernatant
Data file:
Capto adhere
Do you want to view an example of screening and optimization of loading conditions for Capto Adhere? Use our web based application !
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