Development, modeling and optimization of DNA-inorganic hybrid based nanomaterials as signal platform for GMO detection

The use of DNA sequences as a bioreceptor in the design of DNA-based biosensors is a viable alternative to traditional methods for a variety of analyses. Personal glucose meters, which are low-cost and easily accessible biosensors, can indirectly detect a variety of analytes other than glucose, allowing for rapid and more practical analysis. It is possible to use a personal glucose meter to detect sucrose, whose enzymatic product is glucose other than blood sugar when it is hydrolyzed with the invertase enzyme. DNA sequences and the invertase enzyme must be brought together on a suitable platform to accomplish this. It is hoped that by using the CaMV 35S promoter as the model target sequence, a new signaling platform based on DNA-inorganic hybrid-based nanomaterials (DIN) will be developed to determine the GMO amount practically with a personal glucose meter. Magnetite cross-linked invertase aggregates (MCLIA) were prepared for this purpose, and DIN was created by attaching probe DNAs to these aggregates. As a result of mathematical modeling and optimization research studies within the scope of the study, the most ideal structural parameter values are enzyme/MA combination: 0.624 U/mg, glutaraldehyde amount: 5% and experimental parameter values are the pH of reactions: 5, reaction temperature: 50 °C, and reaction time: 45 min. The signal platform's kinetic parameters (Vmax and Km values) for detecting sucrose were determined to be 6625 mg/dl and 226 mM, respectively. Furthermore, the components of the DIN structure were characterized using various techniques, and the interactions of the structural components with each other were determined as a result. After the DIN structure was hybridized with the target sequence (DINH), both structures were incubated with silver ions, and the inhibition difference based on the interactions of single and double-strand DNA sequences with silver was measured using a glucose meter. As a result of the experimental studies with two different flows and three different scenarios, inhibition rates ranging from 20% to 55% in signal values were detected with target DNA binding to the DIN structure. Because the DIN structure is a signal platform, it can be used in future experiments for a variety of purposes (virus, pathogen, genetic diseases, cancer diagnosis, etc.).