Study Of Surface Plasmon Resonance (SPR)-Based Biosensor Immunoassay On Detection Of Medroxy Progesterone Acetate

The residues of veterinary drug in food of animal origin have potential hazards to people's health. Medroxy progesterone acetate (MPA) is a synthetic drug, used for contraception,dysmenorrhea,dysfunctional uterine bleeding, habitual abortion and treatment of cancer. It also used in superovulation, synchronous estrus, promoting growth and restraining rut in animals. But MPA can cause depression, irreversible hypertension, allergies, and even cause breast cancer and infertility. Some countries have even banned use it in animal husbandry, but other countries is still in use. Currently the traditional test methods are mainly high-performance liquid chromatography, capillary electrophoresis and enzyme immunoassay. These methods need complex operation and long time. So we need a new method to monitor MPA better.We purified antibody from ascites were got from mice which were immunity by hybridoma cell line with affinity chromatography technology. Collected the products and then carried on testing the concentration, the purity and the activity. Finally, we obtained three tubes antibody with the concentration 3.21mg/mL, 2.75mg/mL and 1.17mg/mL respectively, and the antibody had good purity and active. We got a good antibody and prepared for the follow-up work.We used indirect-competitive method to test MPA. Synthetic antigen was combined on the surface of a chip. The conditions for immobilizing MPA antigen on the chip were explored. Then we mixed the purified antibody and the inspection sample for a while. Make the mixture flow the surface of the chip. The reactive concentration of antibody and the conditions for regenerating chip were optimized. And the stability and the specificity of the antibody were also evaluated. Results showed that sensitivity was 1ng/mL and specificity was very well. And sample testing results were the same with the kit.The detective time needed for a single sample was very short using the SPR-based biosensor. Thereby it's very suitable for rapid detection on-site and would be have a broad prospect of application.