A two‐step strategy to visually identify molecularly imprinted polymers for tagged proteins

Abstract

A practical and relatively simple method to identify molecularly imprinted polymers capable of binding proteins via the molecular tagging (epitope-like) approach has been developed. In our two-step method, we first challenge a previously obtained anti-tag molecularly imprinted polymer with a small molecule including the said tag of choice (a biotin derivative as shown here or other) connected to a linker bound to a second biotin moiety. An avidin molecule partially decorated with fluorescent labels is then allowed to bind the available biotin derivative associated with the polymer matrix. At the end of this simple process, and after washing off all the low-affinity binding molecules from the polymer matrix, only suitable molecularly imprinted polymers binding avidin through its previously acquired small molecule tag (or epitope-like probe, in a general case) will remain fluorescent. For confirmation, we tested the selective performance of the anti-biotin molecularly imprinted polymer binding it to biotinylated alkaline phosphatase. Residual chemical activity of the enzyme on the molecularly imprinted polymer solid support was observed. In all cases, the corresponding nonimprinted polymer controls were inactive.

Thermo‐sensitive surface molecularly imprinted magnetic microspheres based on bio‐macromolecules and their specific recognition of bovine serum albumin

“…Protein molecularly imprinted polymers (MIPs) are functionally polymeric materials that specifically recognize template proteins . Thus far, MIPs have shown wide application prospects in many fields, such as chemical bionic sensors , protein separation , natural antibody simulation , and drug sustained release . However, the protein bio‐macromolecules of the corresponding MIPs are disadvantaged by weak specificity of the functional groups, large size, and flexible conformations, which challenge their design and preparation .…”

“…Protein molecularly imprinted polymers (MIPs) are functionally polymeric materials that specifically recognize template proteins [1,2]. Thus far, MIPs have shown wide application prospects in many fields, such as chemical bionic sensors [3,4], protein separation [5][6][7], natural antibody simulation [8][9][10], and drug sustained release [11,12]. However, the protein bio-macromolecules of the corresponding MIPs are disadvantaged by weak specificity of the functional groups, large size, Article Related Abbreviations: IF, Imprinting factor; LCST, lower critical solution temperature; MCS, modified chitosan; MIPs, Protein molecularly imprinted polymers; NIPAM, N-isopropylacrylamide; NIPs, Non-imprinted polymer microspheres; Q, Adsorption amount (mg/g); SBMA, sulfobetaine methacrylate; β, selection factor.…”

Preparation and evaluation of temperature and magnetic dual‐responsive molecularly imprinted polymers for the specific enrichment of formononetin

“…Molecular imprinting technology is a predetermined and useful separation and purification method for select specific target molecules by simulating biological antibodies . Molecularly imprinted polymers (MIPs) are prepared by using MIT with tailor‐made recognition sites for a given analyte and its structural analogs .…”

Efficient synthesis of molecularly imprinted polymers with bio‐recognition sites for the selective separation of bovine hemoglobin

“…Molecularly imprinting technology (MIT) is a promising technique for artificial formation of affinity recognition sites in a complicated matrix [10][11][12][13][14][15]. It involves polymerization of functional monomers and crosslinking agent in the presence of a template molecule and subsequent removal of template form the resulting imprinted polymers.…”