A Strep-Tag Imprinted Polymer Platform for Heterogenous Bio(electro)catalysis.

Area of Science:

• Biomaterials Science
• Protein Engineering
• Analytical Chemistry

Background:

• Molecularly imprinted polymers (MIPs) are synthetic receptors with tailored recognition sites.
• Protein MIPs often utilize surface-exposed epitopes as templates, but structural data limitations hinder epitope selection.
• Developing robust MIPs for specific protein recognition remains a challenge.

Purpose of the Study:

• To develop a polyscopoletin-based MIP for recognizing recombinant proteins via the Strep-tag II affinity peptide (Strep-MIP).
• To overcome challenges in selecting suitable epitopes for protein MIPs by using a common affinity tag.
• To create a general and biocompatible platform for immobilizing and utilizing recombinant proteins.

Main Methods:

• Electrosynthesis of Strep-MIP using polyscopoletin.
• Characterization using electrochemistry and surface-sensitive IR spectroscopy.
• Validation through molecular dynamics simulations and binding assays with Strep-tagged enzymes.

Main Results:

• Successful development of a Strep-tag II-imprinted MIP (Strep-MIP).
• Demonstrated recognition and immobilization of two Strep-tagged enzymes: O2-tolerant [NiFe]-hydrogenase and alkaline phosphatase.
• Enzymes retained biocatalytic activity after multiple uses, confirming the platform's efficiency and biocompatibility.

Conclusions:

• Strep-MIP provides a reliable and biocompatible method for imprinting and confining Strep-tagged recombinant proteins.
• This platform facilitates the exploitation of enzymes in various biotechnological applications.
• The approach offers a solution for challenges associated with epitope selection in protein MIP design.