Synthesis and Identification of Artificial Antigen for the Rapid Immunological Detection of Fluoxetine
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Abstract:
Currently, fluoxetine residues in drugs and slimming health foods pose a health problem. Therefore, the synthesis of artificial antigens for fluoxetine was explored, and the structures and specificities of the synthesized hapten and artificial antigen were identified. Fluoxetine reacted with methyl acrylate and underwent hydrolysis to produce the hapten (H-FXT). Subsequently, the active ester method was used to conjugate H-FXT with bovine serum albumin (BSA) and ovalbumin (OVA) separately, to obtain the fluoxetine immunogen (H-FXT–BSA) and coated fluoxetine antigen (H-FXT-OVA), respectively. The H-FXT-BSA immunogen was used to immunize Balb/c mice and the generated anti-serum was analyzed by indirect enzyme-linked immunosorbent assay (ELISA). The artificial antigens, H-FXT-BSA and H-FXT-OVA, were analyzed by ultraviolet (UV) spectroscopy, and calculations showed that the coupling ratios were 14.5:1 and 11.3:1, respectively. The antiserum titers after the immunization of Balb/c mice were all above 30 000, and the half maximal inhibitory concentration was approximately 100 ng / mL. The artificial fluoxetine antigens were successfully synthesized for the first time and anti-fluoxetine antibodies were obtained. This study can provide a key technical basis for further studies involving the establishment of a method for the rapid immunological detection of fluoxetine, including the development of an ELISA kit and other related products.
