DEVELOPMENT OF APTAMER BASED CD62 DIAGNOSTIC KIT

Abstract

Disease-specific biomarkers consider as important tool for the pathological conditions efficient management, including susceptibility determination, diagnosis, and preventive monitoring efficacy. P-selectin is selectively expressed after platelets activation, and it is involved in formation of thrombus, and also in immune response. It is known that serum and plasma p-selectin (CD62) levels increase in some pathological conditions such as heart attack, stroke, some immune diseases and cancer. To date, a lot of methods have been developed to measure p-selectin level. Aptamers use for targeting specific biomarkers on their molecular shape the basis. SELEX (The Systematic Evolution of Ligands by EXponential enrichment) is a process which uses to choose aptamers with high affinity for targeting specific macromolecular. In this study, a new aptamer-based kit was developed to measure CD62. Firstly, aptamers that specifically bind to CD62 were isolated using the SELEX method. Among the used aptamers, Apt-1, Apt-2 and Apt-3 were bound to CD62 protein with high affinity. The Apt-2, which binds with the highest affinity to CD62, the binding constant is -9,6 kcal/mol and dissociation constant (Kd) is 18.15±2.36 nM. We used Bovine serum albumin in the specificity test as a negative control and it was determined that there is no binding between the selected aptamers did not and this protein. The Intra and Inter assay performing showed that the average of Intra-assay was CV % 6,52 and the average of Inter-assay CV % 3,96. As a result, a sensitive, specific, time-saving and low cost kit was developed for the measurement of CD62 with this study.