Determination of Amino Acids and Cyclic Heptapeptides Using Imprinted Proteins
Kirill Yu. Presnyakov1, Polina M. Ilicheva1, Ivan A. Reshetnik1, Mikhail V. Pozharov1, Natalia A. Burmistrova1,2, Pavel S. Pidenko1; 1Saratov State University, Saratov, Russia; 2Saratov Hygiene Medical Research Center of the Federal Government-funded Research Institution, Federal Research Center for Medical and Preventive Health Risk Management Technologies, Saratov, Russia
Abstract
For the first time, a novel approach for the determination of amino acids (Arginine) and heptapeptides (Microcystin—LR) using imprinted proteins as molecular recognition elements in a competitive solid-phase fluorescent assay (CSPA) has been demonstrated. Bovine serum albumin and ovalbumin were used as matrix protein molecules. The efficiency of using dummy templates, specifically an amino acid derivative, in the imprinting process was confirmed.
The resulting imprinted proteins were used to modify the wells of a microtiter plate and the surface of aminated silica (IV) nanoparticles. Application of CSPA enabled achievement of a linear determination range for arginine of 0.7–10 ng/mL (LoQ = 0.5 ng/mL), and for microcystin—LR of 4–60 ng/mL (LoQ = 2 ng/mL). Silica nanoparticles modified with imprinted proteins exhibited significant sorption capacity for arginine, measured at 7.8 mg/g. The developed analytical method was successfully applied for determination of arginine in dietary supplements, demonstrating excellent selectivity and minimal cross-reactivity with other amino acids. Recovery studies yielded an average of 103%, indicating high accuracy and good agreement with concentrations declared by supplement manufacturers.
Furthermore, imprinted proteins synthesized in the presence of the arginine derivative were successfully used for detection of microcystin—LR in standard samples. The proposed IPs-CSPA method represents a promising, sensitive, and selective tool for the analysis of amino acids and toxic peptides in biological and food matrices.
This work was supported by the Russian Science Foundation (project no. 24–73–00250).
Speaker
Kirill Presnyakov
Institute of Chemistry, Saratov State University
Russia
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