A novel analytical probe binding to a potential carcinogenic factor of N-glycolylneuraminic acid by SELEX.

Abstract N -glycolylneuraminic acid (Neu5Gc) is an abundant sialic acid in many mammals and is present in the glycoconjugates of most deuterostome animals. Neu5Gc also occurs in fresh samples of human tumors and fetuses. However, very little is known about the expression level and biologic functions of Neu5Gc due to the limitations of available analytical probes for detection methods. In this study, we first report the development of aptamers specific to Neu5Gc screened by Systematic Evolution of Ligands by Exponential Enrichment (SELEX). After 15 selection rounds, cloning, sequencing and enzyme-linked immuno sorbent assay (ELISA) analysis, 6 different selected aptamers showed specificity for Neu5Gc. Among these 6 aptamers, N8 showed the best half maximal inhibitory concentration (IC 50 ) value (127 ng mL −1 ) and had a relatively high affinity constant (Ka=6.68×10 9  M −1 ). The aptamers selected in this study will provide a novel analytical probe for the development of a biosensor to detect Neu5Gc in tissues and sera from patients with tumors as well as to detect Neu5Gc in animal-derived foods. In addition, the successful aptamer candidate can solve the problem that antibody is difficult to prepare in immunological assays. Thus, the discovery of novel aptamers specific for Neu5Gc is important for developing new methods of detecting Neu5Gc for the diagnosis and prevention of cancer as well as food poisoning.