“Smart” molecularly imprinted monoliths for the selective capture and easy release of proteins

Abstract

A new thermally switchable molecularly imprinted monolith for the selective capture and release of proteins has been designed. First, a generic poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolith reacted with ethylenediamine followed by functionalization with 2-bromoisobutyryl bromide to introduce the initiator for atom transfer radical polymerization. Subsequently, a protein-imprinted poly(N-isopropylacrylamide) layer was grafted onto the surface of the monolithic matrix by atom transfer radical polymerization. Scanning electron microscopy and energy-dispersive X-ray spectroscopy of the cross-sections of imprinted monoliths confirmed the formation of dense poly(N-isopropylacrylamide) brushes on the pore surface. The imprinted monolith exhibited high specificity and selectivity toward its template protein myoglobin over competing proteins and a remarkably large maximum adsorption capacity of 1641 mg/g. Moreover, this "smart" imprinted monolith featured thermally responsive characteristics that enabled selective capture and easy release of proteins triggered only by change in temperature with water as the mobile phase and avoided use of stronger organic solvents or change in ionic strength and pH.

Thermoresponsive hydrophobic copolymer grafted on agar microspheres for all‐aqueous bioseparations

“…The chromatograms in Figure C–E show that the retention times increased in the order as follows: myoglobin (1) < lysozyme (2) < BSA (3) when the proteins were loaded onto the columns at 50°C. This phenomenon is generally in agreement with the relative hydrophobicity of these proteins . Furthermore, the adsorption capacity for the proteins increased with increasing LMA composition at 50°C as a consequence of the change of the hydrophobicity of the agar‐NIPAAm‐ co ‐LMA‐ co ‐AAm column.…”

“…To investigate the hydrophobicity of the prepared microspheres, the temperature-modulated adsorption and elution profiles of proteins mixtures were studied at different temperatures on the agar-NIPAAm-co-LMA-co-AAm column with 1, 3, and 5 mol % LMA compositions. The chromatograms in Figure 5C-E show that the retention times increased in the order as follows: myoglobin (1) < lysozyme (2) < BSA (3) when the proteins were loaded onto the columns at 50 • C. This phenomenon is generally in agreement with the relative hydrophobicity of these proteins [23,24]. Furthermore, the adsorption capacity for the proteins increased with increasing LMA composition at 50 • C as a consequence of the change of the hydrophobicity of the agar-NIPAAm-co-LMA-co-AAm column.…”

Novel functionalized poly(glycidyl methacrylate-co-ethylene dimethacrylate) microspheres for the solid-phase extraction of glycopeptides/glycoproteins

“…2 spectrum a; weak IR bands at 465 and 527 cm −1 are ascribed to C-C skeleton vibration of straight-chain alkanes, and the stronger ones at 758-982 cm −1 are characteristic of CH 3metal groups owing to CH 2 rocking vibration and ring vibration of oxirane groups. The peaks of C-O-C asymmetry stretching vibration appear at 1155-1257 cm −1 [26,39], and the peaks appearing at 1470, 1723, and 2941 cm −1 possibly represent the absorption of the C-H scissor vibration, C=O, and -CH 2 groups, respectively. Upon comparing Fig.…”

“…Recently, Wen et al. designed and prepared a new thermally switchable molecularly imprinted monolith by grafting the pores of PGMA/EDMA for the selective capture and release of proteins . In addition, PGMA/EDMA beads were also applied to enzyme immobilization to improve the performance of the enzyme in terms of storage stability, thermostability, and strong resistance against acid .…”

“…Hao et al presented a new method of surface-initiated atom transfer radical polymerization to design poly(vinyltetrazole)grafted chromatographic packings based on monodisperse PGMA/EDMA beads [25]. Recently, Wen et al designed and prepared a new thermally switchable molecularly imprinted monolith by grafting the pores of PGMA/EDMA for the selective capture and release of proteins [26]. In addition, PGMA/EDMA beads were also applied to enzyme immobilization to improve the performance of the enzyme in terms of storage stability, thermostability, and strong resistance against acid [27].…”