Selective Binding of Inhibitor‐Assisted Surface‐Imprinted Core/Shell Microbeads in Protein Mixtures

Abstract

An innovative approach for molecularly imprinting proteins, i. e., inhibitor‐assisted imprinting was used for addressing the major challenge of protein purification from biotechnologically relevant media. Pepstatin‐assisted pepsin surface‐imprinted microbeads were synthesized as a selective sorbent for solid phase extraction (SPE). The amino‐functionalized porous core was prepared by co‐condensation and post‐grafting strategies. The immobilized inhibitor pre‐organizes the template pepsin with a defined orientation prior to imprinting ensuring enhanced efficiency of the imprinting process. 3‐aminophenylboronic acid (APBA) was used as a functional monomer establishing organic nanoscale films comprising poly(3‐aminophenylboronic acid) (pAPBA) within the pores of the beads in the presence (MIP) and absence (NIP) of pepsin. Next to a detailed characterization of these advanced functional hybrid materials it was demonstrated that such porous surface‐imprinted core/shell beads feature exquisite selectivity for pepsin during individual protein rebinding studies, and more importantly, during competitive rebinding studies in protein mixtures.

Selective binding of matrix metalloproteases MMP-9 and MMP-12 to inhibitor-assisted thermolysin-imprinted beads

“…In contrast to conventional imprinting, the technique of inhibitorassisted imprinting enhances the selectivity of the MIPs by immobilizing an inhibitor prior to the imprinting process. 23,24 The technique of inhibitor-assisted molecularly-imprinting of polymers is based on the copolymerization of functional monomers, an inhibitor and cross-linkers in presence of a template molecule. Thereby binding sites are formed that are complementary in size and shape to the target molecule.…”

“…In the inhibitor-assisted imprinting technique, an inhibitor is immobilized onto the surface of the supporting material, which preorganize the template molecules with a dened orientation prior to the polymerization process. [23][24][25][26][27] The method combines the advantages of surface imprinting strategies and the inhibitor-based affinity interactions. The strategy of the surface imprinting is to place binding sites at or very close to the surface for unhindered access to the binding sites.…”

Core-Shell Imprinted Particles for Adenovirus Binding

“…In addition, successful functionalization was confirmed by XPS analysis (Table 2). For silica-APTES samples, the N 1s peak at 399 eV is visible in the spectra, which is characteristic for the amino group [25,26]. Increase of carbon concentration was monitored during the functionalization, since carbon containing groups were introduced in both steps.…”

Selective Navigation of Bisphenol‐A from Water to a Polarity Tuned Porous Molecularly Imprinted Polymer

“…MIPs differ from the conventional adsorbents by the selectivity and binding capacity of a specific analyte. Therefore, MIPs have been widely studied for various applications such as chromatographic separations, catalysis, drug delivery, membrane separations, sensors and adsorbents for pollutants . The MIPs synthesized in organic solvent exhibit different swelling behavior as well as different size and shape of the cavities when those are employed to adsorb organic pollutants from water and thus often fail to distinguish between target and non‐target molecules .…”