Preparation of molecularly imprinted polymers specific to glycoproteins, glycans and monosaccharides via boronate affinity controllable–oriented surface imprinting

• Published in: Nature protocols Vol. 12; no. 5; pp. 964 - 987
• Main Authors: Xing, Rongrong, Wang, Shuangshou, Bie, Zijun, He, Hui, Liu, Zhen
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.05.2017; Nature Publishing Group
• Subjects: 631/1647/1888; 631/61/2049/974; 639/638/11/872; 639/638/549/941; Acids; Analytical Chemistry; Antibodies; Biological Techniques; Biomarkers; Biomimetics; Biopolymers; Computational Biology/Bioinformatics; Disease; Enzymes; Glycoproteins; Glycoproteins - analysis; Glycoproteins - isolation & purification; Life Sciences; Medical diagnosis; Microarrays; Molecular Imprinting - methods; Monosaccharides - analysis; Monosaccharides - isolation & purification; Organic Chemistry; Polymerization; Polymers; Polysaccharides - analysis; Polysaccharides - isolation & purification; Production processes; Protocol; Reagents; Signal transduction
• ISSN: 1754-2189; 1750-2799; 1750-2799
• DOI: 10.1038/nprot.2017.015

This protocol describes how to produce molecularly imprinted polymers, synthetic receptors that can be produced with antibody-like binding properties. They are easier and cheaper to produce than antibodies and lectins. Molecularly imprinted polymers (MIPs) are materials that are designed to be receptors for a template molecule (e.g., a protein). They are made by polymerizing the polymerizable reagents in the presence of the template; when the template is removed, the material can be used for many applications that would traditionally use antibodies. Thus, MIPs are biomimetic of antibodies and in this capacity have found wide applications, such as sensing, separation and diagnosis. However, many imprinting approaches are uncontrollable, and facile imprinting approaches widely applicable to a large variety of templates remain limited. We developed an approach called boronate affinity controllable–oriented surface imprinting, which allows for easy and efficient preparation of MIPs specific to glycoproteins, glycans and monosaccharides. This approach relies on immobilization of a template (glycoprotein, glycan or monosaccharide) on a boronic-acid-functionalized substrate through boronate affinity interaction, followed by self-polymerization of biocompatible monomer(s) to form an imprinting layer on the substrate with appropriate thickness. Imprinting in this approach is performed in a controllable manner, permitting the thickness of the imprinting layer to be fine-tuned according to the molecular size of the template by adjusting the imprinting time. This not only simplifies the imprinting procedure but also makes the approach widely applicable to a large range of sugar-containing biomolecules. MIPs prepared by this approach exhibit excellent binding properties and can be applied to complex real samples. The MIPs prepared by this protocol have been used in affinity separation, disease diagnosis and bioimaging. The entire protocol, including preparation, property characterization and performance evaluation, takes ∼3–8 d, depending on the type of substrate and template used.