3D Fluorescence Spectroscopy as a Approach to Monitoring of Imprinted Proteins Purification

Abstract

Imprinted proteins (IP) are synthetic analogs of antibodies, IP are produced from proteins initially do not have specificity to target molecules. The process of IP production includes several main stages: protonation of amino acid residues of protein molecule; addition of template molecules and formation of protein–template complex; fixation of the complex by covalent cross-linking; removal of template from protein matrix. The stage of template removal is one of the important and main stages of synthesis. Incomplete removal of template molecules can lead to misleading results in analytical application of IP. The process of removing template molecules and optimization of IP purification methods have not yet been studied.

Fluorescence spectroscopy is one of the simple, fast and highly sensitive methods for studying interactions in various systems. The use of 3D fluorescence spectroscopy allows us to obtain more information about the protein-template system compared to other spectroscopic methods. In this work, we used bovine serum albumin and glucose oxidase as matrix protein molecules, and zearalenone and its structural analog 4-hydroxycoumarin as template molecules. Dialysis was used to remove low molecular weight templates. Visualization of the results of 3D fluorescence spectroscopy in the form of excitation-emission matrixes allowed us to confirm the almost complete removal of templates from the protein matrix.

Speaker

Kirill Presnyakov
Saratov State University, Institute of Chemistry
Russia

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