Preparation of a monolithic column for weak cation exchange chromatography and its application in the separation of biopolymers

Abstract

Original PaperPreparation of a monolithic column for weak cation exchange chromatography and its application in the separation of biopolymers A procedure for the preparation of a monolithic column for weak cation exchange chromatography was presented. The structure of the monolithic column was evaluated by mercury intrusion. The hydrodynamic and chromatographic properties of the monolithic column -such as back pressures at different flow rates, effects of pH on protein retention, dynamic loading capacity, recovery, and stability -were determined under conditions typical for ion-exchange chromatography. The prepared monolithic column might be used in a relatively broad pH range from 4.0 to 12.0 and exhibited an excellent separation to five proteins at the flow rates of both 1.0 and 8.0 mL/min, respectively. In addition, the prepared column was first used in the purification and simultaneous renaturation of recombinant human interferon gamma (rhIFN-c) in the extract solution with 7.0 mol/L guanidine hydrochloride. The purity and specific bioactivity of the purified rhIFN-c in only one chromatographic step were obtained to be 93% and 7.8610 7 IU/mg, respectively.

Weak cation‐exchange monolithic column for capillary liquid chromatography of peptides and proteins

“…However, the decrements for different proteins were not similar. This behavior has also been previously described 32, 44. Ribonuclease A eluted after α‐chymotrypsinogen A and cytochrome c at pH 6.0, whereas it eluted between α‐chymotrypsinogen A and cytochrome c at pH 7.0 and 8.0, but before them at pH 9.0.…”

“…Separation of isomers of aspartyl peptides was achieved in the CEC mode using the weak cation‐exchange column, whereas no separation was obtained by capillary zone electrophoresis (CZE) and HPLC. Other methods to prepare monolithic columns for WCX were also reported, such as ring‐opening metathesis polymerization 30, grafting 31 and post‐modification 32–34.…”

Preparation of high-capacity strong cation exchange membrane for protein adsorption via surface-initiated atom transfer radical polymerization

“…To investigate this nonspecific protein adsorption, IPA was used as an additive in the mobile phase to detect any hydrophobic adsorption between the solutes and the membrane material [37]. If hydrophobic interactions exist, adding a small amount of IPA to the buffer should reduce the adsorption significantly.…”

Macroporous Monolithic Polymers: Preparation and Applications

“…Based on poly(GMA-co-DVB) monoliths, DEAE or QA functionalities were introduced to provide WAX or SAX stationary phases for the separation of oligonucleotides, or oligonucleotides and nucleosides, respectively [66,67]. Wei et al [68] presented the preparation of a poly(GMA-co-EGDMA)-based monolith column for weak cation exchange chromatography and its use in the separation of biopolymers. Iminodiacetic acid (IDA)-type adsorbent was prepared by covalent coupling of IDA to the monolithic macroporous poly(GMA-co-EGDMA).…”