3/21/2023 0 Comments Capto adhere resin![]() This study demonstrated that mixed mode resins can be readily integrated into commercial antibody platform processes when additional chromatographic abilities are required.Īntibody Fragment removal Mixed mode resin.Ĭopyright © 2017 The Author(s). This study demonstrated that mixed mode resins can be readily integrated into commercial antibody platform processes when additional chromatographic abilities are required. Additionally, a case study is presented detailing the optimization of fragment removal using Capto adhere resin to achieve purity and yield targets in a manufacturing facility. Additionally, a case study is presented detailing the optimization of fragment removal using Capto adhere resin to achieve purity and yield targets in a manufacturing facility. Sequence analysis showed that under separation conditions, a hydrophobic proline patch and hydrogen bonding serine and threonine residues mediate the hinge interaction with the Capto adhere ligand. The labeling identified the antibody hinge and light chain regions as mediating the fragment separation. The site of interaction between the LHF and mixed mode resin was determined by chemical labeling of lysine residues with sulfo-NHS acetate. Therefore, it was discovered that the purification is the result of a mixed mode phenomena dominated by hydrophobic interaction and hydrogen bonding effects. Further decreases in LHF separation were seen upon incubation with urea and arginine. Capto MMC combines agarose base matrix developments with innovative ligand chemistry. The addition of ethylene glycol decreased LHF removal by half. Capto MMC MULTIMODAL RESIN Capto MMC is a multimodal cation exhanger in the Capto family of BioProcess resins for fast, efficient and cost-effective protein purification. The second experimental series studied the impact of phase modifiers, ethylene glycol, urea, and arginine on the mixed mode mediated removal. ![]() Removal of greater than 75 of LHF population occurred at pH 8 and low conductivity. ![]() Both single mode anion exchange and hydrophobic interaction resins failed to separate LHF. Capto adhere, HEA Hypercel, and PPA Hypercel anion exchange/hydrophobic interaction mixed mode resins were evaluated for their fragment removal capabilities and found to separate large hinge IgG1 antibody fragment (LHF) from monomer. The first experimental series consisted of comparison to chromatographic behavior on corresponding single mode resins. The mechanism of fragment removal was investigated in two series of experiments. Removal of greater than 75% of LHF population occurred at pH 8 and low conductivity. Capto adhere, HEA Hypercel, and PPA Hypercel anion exchange/hydrophobic interaction mixed mode resins were evaluated for their fragment removal capabilities and found to separate large hinge IgG1 antibody fragment (LHF) from monomer. Efforts to increase monoclonal antibody expression in cell culture can result in the presence of fragmented species requiring removal in downstream processing. Capto adhere can remove key impurities in a single step, allowing the design of a two-step process together with a protein A media (e.g. ![]()
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