Preparation of Monoclonal Antibodies for Development of Indirect Competitive ELISA for Salidroside Detection

  Objective  To prepare monoclonal antibody of salidroside for the development of an indirect competitive enzyme linked immunosorbent assay (ic-ELISA) methodology to accurately and rapidly detect salidroside in medicinal materials.

  Method  Balb/c mice of 6-8 weeks old were immunized with the prepared salidroside-BSA. Specificity of the anti-serum was determined by ELISA. Spleen cells from the mice with positive result were fused with SP2/0 myeloma cells. Monoclonal hybridoma cells were screened by indirect ELISA with a limited dilution. Ascitic antibodies were induced and prepared from the positive cell line. Specific antibody against salidroside was used to establish ic-ELISA detection method that was verified by its specificity, precision and recovery rate on a known standard.

  Result  The sensitivity of ic-ELISA was 49.33 ng·mL^-1 with a liner range of 4.07-598.45 ng·mL^-1. The detection limit of the method was 1.77 ng·mL^-1. The newly developed method detected salidroside in spiked samples with a recovery rate of 94.3% within the designed range. The relative standard deviation of the measurements was less than 4.5% on both intra- and inter-day assays. The determination by ic-ELISA agreed with that obtained by HPLC.

  Conclusion  The newly established ic-ELISA method based on the salidroside monoclonal antibody was considered appropriate for the detection.