The presented project combines reproductive biology and nanotechnology. A nano-purification method will be developed to be applied during the freezing of bull sperm, aiming to increase the quality of the frozen bull sperm and the fertility capability of the post-thaw sperm. In this context, target-specific magnetic nanoparticles (TSMNs) will be created by binding Lectin/PNA, Lectin/PSA, Annexin V, and Anti-Ubiquitin biomarkers to Fe3O4 nanoparticles using a binding agent during equilibration. These TSMNs, magnetized at the end of equilibration, will attach to damaged spermatozoa via their biomarkers, thus segregating damaged and non-motile spermatozoa from healthy ones. Damaged spermatozoa separated by this method will not enter the freezing protocol, ensuring that only sperms with the desired spermatozoological characteristics are frozen. During freezing, the prevention of ROS formation from damaged spermatozoa is expected to result in a more successful freezing process, leading to post-thaw sperm with high viability and motility. The project is designed in two stages: a main project and a sub-project. The main project consists of two stages, where in the first stage, different doses and sizes of Fe3O4 nanoparticles will be tested. The two best doses and sizes showing no toxic effects on sperm and optimal binding to sperm will be determined. In the second stage of the project, biomarkers will be attached to the determined doses and sizes of nanoparticles to form TSMNs, which will be added to the sperm freezing protocol. Throughout the project, the success of the nano-purification process will be measured using computer-assisted sperm analysis (CASA), flow cytometry, and electron microscopic imaging techniques. The sub-project will evaluate the advantages and disadvantages of the method applied in the project compared to other sperm purification methods and will determine whether nano-purified sperm can be successful in artificial insemination applications. The planned study is unique in that it will be the first to experiment with Fe3O4 nanoparticles of three different sizes and doses, and therefore different structures, not previously investigated in the literature. Also, unlike previous studies, by simultaneously using four different biomarkers to detect all possible damages that may occur in sperm, this study will allow for the comparison of damage causes and the interpretation of meaningful results between the study group and the control group. The data obtained and the successful purification of sperm at the conclusion of the project will contribute to Turkish animal husbandry and Reproduction Science under the call program "Development of New Systems, Models, and Methods Aimed at Improving Yields in Terms of Quality and Quantity."