![]() The differences in chromatographic behaviour of individual deoxynucleotides as well as small single-stranded and double-stranded DNA molecules have been examined for two resins from the Capto family: Capto Adhere and Capto Q ImpRes. Ultra C8 Columns (USP L7), Particle Size 5 μm, Length 250 mm, ID 4.6 mm, Restek (CAT#: STEM-C-0822-LC) High-resolution intermediate purification and polishing based on the well-established Capto platform with traditional ligands Shop Cytiva HiScreen Capto Cytiva HiScreen Capto Q ImpRes and Capto SP ImpRes Capto SP ImpRes Ionic Capac.: 0.16-0.Capto MMC ImpRes is a chromatographic medium based on a multimodal cation exchange ligand (Fig 1). Inertsil ODS-SP Cartridge Guard Column E, Replacement Cartridge E (2/pk) & Holder Set, Particle Size 3µm, I.D.The ligand constitutes a hydrophobic part, a weak cation exchange group, and groups that can promote hydrogen bonds. Rtx-1 Columns (Fused Silica), Length 15 m, ID 0.32 mm, df 0.25 μm, Temp.Ĭapto MMC ImpRes comprises small ( 40 m) beads relative to the related multimodal cation exchange medium, Capto MMC ( 75 m).Capto Q ImpRes: Strong anion (quaternary amine group) exchange BioProcess medium Capto SP ImpRes: Strong cation (sulfonate group) exchange BioProcess medium. Inertsil ODS-HL Guard Columns For UHPLC, Replacement Cartridge (2/pk) & Holder Set For UHPLC, Particle Size 3µm, I.D.HiTrap column prepacked with Capto Q ImpRes resin. InertSustainSwift C18 Capillary Columns, Particle Size 5µm, I.D.Designed to offer quality and flexibility in process-scale applications. Scaling up allows for high resolution combined with quick flow rates. All rights reserved.Capto Q ImpRes resin couples a quarternary ammonium (Q) strong anion exchanger to a chemically modified, high-flow agarose matrix. ![]() 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). 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. The addition of ethylene glycol decreased LHF removal by half. The second experimental series studied the impact of phase modifiers, ethylene glycol, urea, and arginine on the mixed mode mediated removal. Both single mode anion exchange and hydrophobic interaction resins failed to separate LHF. 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. ![]()
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