Abstract
Three-dimensional printing applications in separation science are currently limited by the lack of materials compatible with chromatographic operations and three-dimensional printing technologies. In this work, we propose a new material for Digital Light Processing printing to fabricate functional ion exchange monoliths in a single step. Through copolymerization of the bifunctional monomer [2-(acryloyloxy)ethyl] trimethylammonium chloride, monolithic structures with quaternary amine ligands were fabricated. The novel formulation was optimized in terms of protein binding and recovery, microporous structure, and its swelling susceptibility by increasing its cross-link density and employing cyclohexanol and dodecanol as pore forming agents. In static conditions, the material demonstrated a maximum binding capacity of 104.2±10.6mg/mL for bovine serum albumin, in line with commercially available materials. Its anion exchange behavior was validated by separating bovine serum albumin and myoglobin on a monolithic bed with Schoen gyroid morphology. The same column geometry was tested for the purification of C-phycocyanin from clarified as well as cell-laden Arthrospira platensis feedstocks. This represents the first demonstration of one-step printed stationary phases to capture proteins directly from solid-laden feedstocks. We believe that the material presented here represents a significant improvement towards implementation of three-dimensional printed chromatography media in the field of separation science.
Highlights
Chromatographic separations currently rely on randomly packed spherical adsorber beads, with slurry packing procedures being the only economical and feasible method for column packing
digital light processing (DLP) 3D printing materials are complex mixtures composed of photopolymerisable monomers and crosslinkers forming the polymeric network, photoinitiators required to start the photopolymerisation reaction and photoabsorber to increase the printing resolution [10,11]
Optimization of the parent material composition enabled overcome a number of hurdles associated with previous “one-step” 3D printable materials for chromatography, including improved protein binding through selection of appropriate ligand density, enhanced elution characteristics for higher protein recoveries, improved mechanical properties, more open porous microstructure to favour mass transfer within the stationary phase, and a more “customer-oriented” aesthetic appearance
Summary
Chromatographic separations currently rely on randomly packed spherical adsorber beads, with slurry packing procedures being the only economical and feasible method for column packing. With the introduction of 3D printers, it is feasible to fabricate stationary phases with ordered three-dimensional morphology according to digital models [3]. The suitability of 3D printing to create ordered structures for chromatographic applications has been previously demonstrated. The commercial materials employed did not allow for the testing of protein separation due to their lack of suitable chromatographic ligands. McDonald et al [5] employed a commercial material (Veroclear-RGD810, Stratasys Ltd.) for 3D printing that displayed a net negative surface charge to enable separation of a range of test components, such as dyes and proteins. According to a different preparation method, commercial materials for 3D printing can be used to fabricate sacrificial moulds, which are infused with a traditional material for chromatography such as porous agarose or cellulose hydrogels, followed by chemical modification for the introduction of the chromatographic ligand [6]. The onestep fabrication method proposed was based on the bifunctional monomer 2-(Acryloyloxy)ethyl] trimethylammonium chloride (AETAC), bearing a positively charged quaternary amine (QA) group as well as a 3D printable acrylate group in its structure (
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