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

Chen, Xin; Tolley, H. Dennis; Lee, Milton L.

doi:10.1002/jssc.200900255

Cited by 23 publications

(13 citation statements)

References 60 publications

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“…The use of bifunctional monomers has been explored in IEX monoliths by Milton Lee's group. Fully functional porous monoliths were prepared in a single step synthesis via UV initiated polymerisation by copolymerising the crosslinker PEGDA with bifunctional monomers containing sulfonic acid [23,24], phosphoric acid [25], carboxylic acid [26] or amine groups [27]. Porosities were created by the solvents present in the reacting mixture.…”

Section: Lack Of Suitable Materials For Chromatographic Separations Tmentioning

confidence: 99%

Direct 3D printing of monolithic ion exchange adsorbers

Simon

Dimartino

2019

Journal of Chromatography A

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Monolithic adsorbers with anion exchange (AEX) properties have been 3D printed in an easy one-step process, i.e. not requiring post-functionalization to introduce the AEX ligands. The adsorber, 3D printed using a commercial digital light processing (DLP) printer, was obtained by copolymerisation of a bifunctional monomer bearing a positively charged quaternary amine as well as an acrylate group, with the biocompatible crosslinker polyethylene glycol diacrylate (PEGDA). To increase the surface area, polyethylene glycol was introduced into the material formulation as pore forming agent. The influence of photoinitiator (Omnirad 819) and photoabsorber (Reactive Orange 16, RO16) concentration was investigated in order to optimize printing resolution, allowing to reliably 3D print features as small as 200 µm and of highly complex Schoen gyroids. Protein binding was measured on AEX adsorbers with a range of ligand densities (0.00, 2.03, 2.60 and 3.18 mmol/mL) using bovine serum albumin (BSA) and c-phycocyanin (CPC) as model proteins. The highest equilibrium binding capacity was found for the material presenting the lowest ligand density analysed (2.03 mmol/mL), adsorbing 73.7 ± 5.9 mg/mL and 38.0 ± 2.2 mg/mL of BSA and CPC, respectively. This novel 3D printed material displayed binding capacities in par or even higher than commercially available chromatographic resins. We expect that the herein presented approach of using bifunctional monomers, bearing commonly used chromatography ligands, will help overcome the material limitations currently refraining 3D printing applications in separation sciences.

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Section: Lack Of Suitable Materials For Chromatographic Separations Tmentioning

confidence: 99%

Direct 3D printing of monolithic ion exchange adsorbers

Simon

Dimartino

2019

Journal of Chromatography A

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“…Polymer‐based monoliths are widely utilized in capillary LC (cLC) and CEC as stationary phase . Polymer‐based chiral stationary phases represent a new attractive topic of research in the chiral separation science.…”

Section: Introductionmentioning

confidence: 99%

Enantioseparation of aromatic amino acids using CEC monolith with novel chiral selector, N‐methacryloyl‐l‐histidine methyl ester

et al. 2013

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A new type of polymethacrylate-based monolithic column with chiral stationary phase was prepared for the enantioseparation of aromatic amino acids, namely D,L-phenylalanine, D,L-tyrosine, and D,L-tryptophan by CEC. The monolithic column was prepared by in situ polymerization of butyl methacrylate (BMA), N-methacryloyl-L-histidine methyl ester (MAH), and ethylene dimethacrylate (EDMA) in the presence of porogens. The porogen mixture included DMF and phosphate buffer. MAH was used as a chiral selector. FTIR spectrum of the polymethacrylate-based monolith showed that MAH was incorporated into the polymeric structure via in situ polymerization. Some experimental parameters including pH, concentration of the mobile phase, and MAH concentration with regard to the chiral CEC separation were investigated. Single enantiomers and enantiomer mixtures of the amino acids were separately injected into the monolithic column. It was observed that L-enantiomers of aromatic amino acids migrated before D-enantiomers. The reversal enantiomer migration order for tryptophan was observed upon changing of pH. Using the chiral monolithic column (100 μm id and 375 μm od), the best chiral separation was performed in 35:65% ACN/phosphate buffer (pH 8.0, 10 mM) with an applied voltage of 12 kV in CEC. SEM images showed that the chiral monolithic column has a continuous polymeric skeleton and large through-pore structure.

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“…Methanol has been a common solvent used in preparation of monoliths containing PEG functional groups due to its good solubility 24, 25, 38, 39. Therefore, methanol was chosen as the initial porogen solvent to prepare the poly(CEA‐ co ‐PEGDA) monolith.…”

Section: Resultsmentioning

confidence: 99%

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

Chen¹,

Tolley²,

Lee³

2011

J of Separation Science

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A stable poly(2-carboxyethyl acrylate-co-poly(ethylene glycol) diacrylate) monolith was synthesized inside a 75-mm id capillary by direct in situ photo-initiated polymerization in a binary porogenic solvent consisting of methanol and ethyl ether. The resulting monolith was evaluated for weak cation-exchange capillary liquid chromatography of peptides and proteins. A high dynamic binding capacity of 72.7 mg lysozyme per cm 3 column volume was measured. Fast mass transfer was demonstrated by steep breakthrough curves. The resulting monolith exhibited negligible hydrophobicity, leading to good separation of peptides and proteins. Peak capacities of 11 for peptides with a 10-min salt gradient and 39 for proteins with a 20-min salt gradient were measured. An efficiency of 37 000 plates/ m for proteins was obtained under isocratic conditions. The effects of functional group concentration, porogenic solvent composition, mobile phase pH, salt gradient rate, and hydrophobicity on the retention of analytes were investigated. Good run-to-run relative standard deviation (RSD) o1.93% and column-to-column RSD o4.63% were achieved.

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

Cited by 23 publications

References 60 publications

Direct 3D printing of monolithic ion exchange adsorbers

Direct 3D printing of monolithic ion exchange adsorbers

Enantioseparation of aromatic amino acids using CEC monolith with novel chiral selector, N‐methacryloyl‐l‐histidine methyl ester

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

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