Metal Organik Kafes Yapılarının Diol İçeren Biyomolekül Tayini ve İzolasyonu İçin Kullanılabilirliğinin İncelenmesi

Abstract

The use of metal-organic frameworks (MOFs) in proteomics applications makes a significant contribution to materials science research due to their versatile properties such as high surface area and ease of modification. Since they have higher selectivity, sensitivity and reproducibility than traditionally used materials, they are used as affinity sorbents to enrich diol-containing biomolecules such as glycoproteins, saccharides and nucleosides. Since the levels of diol containing biomolecules reflect the physiological state of humans and are therefore important in disease diagnosis, these biomolecules play an active role in physiological pharmacology studies. Therefore, it is of great importance to design and develop sorbents with high adsorption capacity for diol containing biomolecules. In this study, zirconium aminobenzenedicarboxylate-based MOF (UiO-66-NH2) material functionalized with 4-formylphenylboric acid (FPBA) as a new boronate affinity material (FPBA-UiO-66-NH2) for selective enrichment and elution of biomolecules at appropriate pH (pH=8). As a result of the affinity between boronic acid and diol, a high adsorption capacity of 80 mg/g was achieved for β-NAD, which was selected as the biomolecules with high selectivity and reusability. The adsorbed β-NAD was easily isolated in neutral pH buffer medium with quantitative desorption rate.

In the last few years, colorimetric biosensing has attracted considerable attention in biological sciences and analytical chemistry. This method has the advantages of speed, low-cost equipment, simplicity, low cost and does not require any complex instrumentation. MOFs are excellent peroxidase-like mimics thanks to their controllable preparation methods, tunable structures and activities and high stability. In recent years, studies on biosensing applications of MOFs have gained momentum. Within the scope of the thesis, the MOF-based nanozyme developed for the colorimetric detection of diol containing biomolecule exhibited high peroxidase catalytic activity. The synthesized material catalyzed the oxidation of colorless o-phenylene diamine (OPDA) to the yellow product (referred to as oxOPDA), and this change led to a significant change in the adsorption value at 416 nm. This method was applied for the selective determination and quantification of β-NAD in buffer solution. It was observed that the calibration graph obtained under optimum conditions was linear in the range of 0-0.75 mM. The limit of detection (LOD) of the method was calculated as 0.033 mM (taking S/N=10). All results show that the developed boronic acid group-functionalized MOF material is a promising material for the isolation and determination of diol containing biomolecules.