Immunological kit for detecting florfenicol
The technology of florfenicol and reagent kit is applied in the field of immunological reagent kit for detecting florfenicol, which can solve the problems of complex and expensive instruments, professional personnel required for operation, and high detection cost, and achieve wide application prospects, low price, The effect of high sensitivity
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Embodiment 1
[0064] Embodiment 1 florfenicol complete antigen
[0065] Florfenicol complete antigen is a conjugate of a Florfenicol derivative and a carrier protein, wherein the Florfenicol derivative is abbreviated as FFOA, and its chemical formula is as follows:
[0066]
[0067] The carrier protein is bovine serum albumin (BSA) or chicken ovalbumin (OVA).
[0068] The obtained complete antigen FFOA-BSA is used as the immunogen for the preparation of the camelid antibody, and the complete antigen FFOA-OVA is used as the coating agent constructed by the florfenicol enzyme-linked immunoassay method.
Embodiment 2
[0069] Example 2 Preparation of Florfenicol Alpaca Polyclonal Antibody
[0070] (1) Mix the florfenicol complete antigen FFOA-BSA and Freund's complete adjuvant in equal proportions and fully emulsify the alpaca by subcutaneous injection in the neck and back at a dosage of 750 μg / time;
[0071] (2) Boost immunization on days 22, 36, 50, and 64 respectively (mix FFOA-BSA and Freund's incomplete adjuvant in equal proportions and fully emulsify them, and immunize alpacas by subcutaneous injection at multiple points on the back, with a dosage of 500 μg / time), a total of 4 booster immunizations, with an interval of 2 weeks.
[0072] (3) On the 71st day, 50 mL of whole blood was collected from the neck of the alpaca, placed in a water bath at 37°C for 30 minutes, centrifuged at 12,000 rpm at 4°C for 30 minutes, and the supernatant collected was Florfenicol Alpaca Cloned antibody, aliquoted and stored at -20°C.
[0073] The alpaca was immunized with florfenicol complete antigen F...
Embodiment 3
[0074] Example 3 Preparation of Florfenicol Bactrian camel polyclonal antibody
[0075] (1) The florfenicol complete antigen FFOA-BSA and Freund's complete adjuvant were mixed in equal proportions and fully emulsified to immunize Bactrian camels by multi-point injection in the neck and back, with a dose of 500 μg / time;
[0076] (2) Immunization was boosted on days 15, 29, 43, 57, and 71 (mix FFOA-BSA with Freund's incomplete adjuvant in equal proportions and fully emulsify them, and immunize both sides by subcutaneous injection at multiple points on the neck and back). humpback camel, the dosage is 500μg / time), a total of 5 times of booster immunization, the immunization interval is 2 weeks.
[0077] (3) On the 78th day, 100 mL of whole blood was collected from the neck of the Bactrian camel, placed in a water bath at 37°C for 30 minutes, centrifuged at 12,000 rpm at 4°C for 30 minutes, and the supernatant collected was the florfenicol doublet Camel polyclonal antibody, aliqu...
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