Understanding this behavior has implications in the design of gradient elution since the gradient slope impacts protein recovery. Elution with a reverse gradient is also demonstrated at low salt concentrations for these proteins. High Performance are members of the Cytiva range of hydrophobic interaction chromatography (HIC). One of the mAbs and lysozyme exhibit U-shaped retention factor curves and for each, we determine the critical gradient slope beyond which 100% recovery is no longer possible. Phenyl Sepharose High Performance and Butyl Sepharose. Experimental results are presented for two different monoclonal antibodies and for lysozyme on Capto Phenyl (High Sub) resin. Capto Phenyl (High sub) and Capto Butyl are hydrophobic interaction chromatography (HIC) resins used in the capture and intermediate stages of protein. We show that the normalized gradient slope determines whether the protein elutes in the gradient, partially elutes, or is trapped in the column. Two dynamic models, one based on the local equilibrium theory and the other based on the linear driving force approximation, are presented. For these conditions, a steep gradient can overtake the protein in the column, causing it to rebind. During gradient elution the salt concentration is gradually decreased from a high value thereby reducing the retention factor and increasing the protein chromatographic velocity. Capto Core 700 GE Healthcare Capto Phenyl (Hi-Sub) GE Healthcare Capto Q GE Healthcare Cellufine Q-r JNC Corporation CHT Ceramic Hydroxyapatite Bio-Rad Laboratories CHT Ceramic Hydroxyapatite Type 1 Bio-Rad Laboratories Fractogel EMD TMAE (S) MiliporeSigma Fractogel EMD S0 3 MiliporeSigma Hi C Capto Phenyl (High-Sub. In some cases, however, protein retention also increases at low salt concentrations resulting in a U-shaped retention factor curve. In general, an increase in salt concentration drives protein partitioning to the hydrophobic surface while a decrease reduces it. Protein retention in hydrophobic interaction chromatography is described by the solvophobic theory as a function of the kosmostropic salt concentration.
0 kommentar(er)
