Synthesis of Tenuazonic Acid Artificial Antigens and Preparation of an Anti-tenuazonic Acid Polyclonal Antibody

Abstract:

The oximation reaction was used to derivatize tenuazonic acid (TA), and a connecting arm was introduced to synthesize the hapten, TAO. The TAO-BSA immunogen and TAO-KLH coating antigen conjugates were prepared by the active ester method. After dialysis purification, UV spectrum scanning, Fourier transform infrared (FT-IR) spectroscopy, and animal immunization experiments were used to test and verify the synthesis products. The coupling ratios of the two conjugates were 18.3:1 and 38.7:1. Balb/c mice were immunized by TAO-BSA and a polyclonal TA antibody was obtained from the antiserum. The sensitivity of the mouse antiserum was measured by indirect competitive ELISA. The optimum concentrations of the coating antigen and antibody dilutions were determined by the chessboard method. The sensitivity, as measured by IC50, was 335.55 ng/mL. The detection limit (LOD) was 19.00 ng/mL, and the linearity range was 200.00~1563.00 ng/mL.