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

WANG Wei; HU Li-xin; FU Guang-hua; ZHANG Hai-li

doi:10.19303/j.issn.1008-0384.2019.05.013

Volume 34 Issue 5

Aug. 2019

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WANG Wei, HU Li-xin, FU Guang-hua, ZHANG Hai-li. Preparation of Monoclonal Antibodies for Development of Indirect Competitive ELISA for Salidroside Detection[J]. Fujian Journal of Agricultural Sciences, 2019, 34(5): 595-599. doi: 10.19303/j.issn.1008-0384.2019.05.013

Citation:

WANG Wei, HU Li-xin, FU Guang-hua, ZHANG Hai-li. Preparation of Monoclonal Antibodies for Development of Indirect Competitive ELISA for Salidroside Detection[J]. Fujian Journal of Agricultural Sciences, 2019, 34(5): 595-599. doi: 10.19303/j.issn.1008-0384.2019.05.013

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

doi: 10.19303/j.issn.1008-0384.2019.05.013

1.
Linyi Institute for Food and Drug Control, Linyi, Shandong 276000, China
2.
Heze University, Heze, Shandong 276000, China

Received Date: 2018-12-27
Rev Recd Date: 2019-04-08
Publish Date: 2019-05-28

Abstract

Abstract

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.
- salidroside,
- monoclonal antibody,
- indirect competitive enzyme-linked immunosorbent assay

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References(19)

References

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