Tuzaklamalı İyon Hareketliliği Kütle Spektrometrisi ile İnsan İnsülini ve Analoglarının Analizi

Abstract

Insulin is very important for human life and highly sensitive to environmental factors. The insulin hormone that regulates the sugar in the blood is also involved in protein and fat metabolism. It can easily undergo structural changes according to its functions. The slightest change in the structure of insulin hormone has an important effect on the physiological and pharmacological activities of this hormone. The monomer form of insulin with many oligomeric structures is the form in which it is biologically active. Analyzing insulin structure and functions under different conditions is important to provide better and improved treatment methods. The need for structural knowledge in studies with biomolecules is increasing. Because a single protein, depending on the form of folding can have many different functions and features. Ion Mobility Spectrometry (IMS), which gives information about the three-dimensional structure of an ion, becomes a device that gives the most accurate and reliable information about the conformation properties of the molecule when combined with mass spectrometry (MS), which is a very powerful analytical technique for characterizing the molecule. The IMS separates ions according to the size of the gas phase (Collision Cross-Section, CCS) and their shape, while the MS separates them according to the mass / charge ratio. With the MS system combined with the IMS (IM MS), it is not only the mass examined but also the size and shape of the molecule. Trapped Ion Mobility Mass Spectrometry (TIMS MS) technique, which is named according to the way of separating the ions, offers the advantages of rapid analysis of different types of samples, high sensitivity, better resolving power. In this thesis, the conformational properties of human insulin and its analogs in various solvents, different voltages and temperatures were investigated by the technique of trapped ion mobility mass spectrometry. The results obtained in the thesis study, identify the conformational properties of the insulin molecule, analyze the structure-function relationships, distinguish the analogues from each other and ensure that they can be defined with high precision and accuracy. The effect of the solvent on the conformation of insulin hormone is considerably greater than the effect of the instrument parameters, ion source temperature and capillery voltage. This study, which we have detailed information about the conformation of insulin hormone, will lead all studies to be done with this hormone.