Designing of L-proline specific nanoparticles for L-proline detection from human serum

Processes such as the detection and quantification of amino acids hold significant importance in biochemical analyses, particularly in the diagnosis of metabolic disorders. Among these amino acids, L-Proline has been identified as having significant relevance to various metabolic disorders in living organisms, particularly in human. Hyperprolinemia is a metabolic disorder that arises when the L-proline molecule is not effectively broken down due to deficiencies in proline oxidase or pyrroline-5 carboxylate dehydrogenase enzymes, resulting in an accumulation of L-Proline within the body. In individuals with Hyperprolinemia, there is a noticeable increase in L-Proline levels in both blood and urine, underlining the importance of accurately measuring and monitoring these levels in body fluids. To address the challenge of recognizing target molecules, such as proteins, peptides, amino acids, or ions, in complex environments with high selectivity, molecular imprinting emerges as a dependable technique that accomplishes this in a single step. In this study, molecularly imprinted nanoparticles that can selectively recognize the L-Proline molecule were synthesized. Hydroxyethyl methacrylate (HEMA) based nanoparticles were synthesized via emulsion polymerization technique and were characterized by scanning electron microscopy, zeta-sizer particle size analysis, surface area calculations, and Fourier Transform Infrared Spectroscopy (FTIR). Based on zeta-sizer analysis, the estimated diameters of Pro-MIP and NIP nanoparticles were determined to be approximately 27.51 nm and 20.66 nm, respectively. The adsorption of L-Proline onto nanoparticles from aqueous solutions was investigated in a batch system, and the maximum L-Proline adsorption capacity was determined to be 26.58 mg/g for Pro-MIP and 4.65 mg/g for NIP. The selectivity of L-Proline imprinted nanoparticles was successfully confirmed via Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS), in the presence of competing molecules (L-Histidine and L- Phenylalanine). Finally, Pro-MIPs were subjected to repeated adsorption-desorption cycles and it was demonstrated that Pro-MIPs can be used up to 10 times without significant decrease in selective adsorption capacity.