Artificial antigen and antibody for melamine residual immunoassay and preparation method thereof
A technology of melamine and artificial antibodies, applied in the direction of serum immunoglobulin, animal/human protein, material inspection products, etc., can solve the problems of high price, achieve high similarity, good application prospects, avoid synthetic reactions and remove impurities The effect of the process
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Embodiment 1
[0024] Example 1: Preparation and Identification of Melamine Artificial Antigen (Immune Antigen and Coating Antigen)
[0025] 1. Synthesis of melamine artificial antigen
[0026] Add 41.5-42.5 mg of hapten, 33.5-34.5 mg of NHS, and 61.5-62.5 mg of DCC into 2 mL of DMF, and stir overnight at 4°C. The next day, the mixture was centrifuged, the precipitate was discarded, and the supernatant was absorbed for later use. Dissolve 112.5-113.0 mg of BSA and 74.5-75.0 mg of OVA in 8 mL of 0.01 mol / L, pH=7.4 PBS, divide the obtained supernatant into two parts and slowly add the two solutions while stirring, at 4 °C Stir slowly overnight. Afterwards, the reaction solution was transferred to the treated dialysis bag, collected after thorough dialysis in PBS, subpackaged, and stored at -20°C.
[0027] 2. Identification of melamine artificial antigen
[0028] Ultraviolet spectroscopy is used for identification. After the carrier protein and the hapten are coupled, the product will prese...
Embodiment 2
[0029] Embodiment 2: Preparation and specific identification of melamine antibody
[0030] 1. Preparation of melamine antibody
[0031] (1) Immunization Three-month-old healthy, male New Zealand white rabbits with a weight of about 1.5 kg were selected and raised in a standard experimental animal room for observation for 3 days, and then immunized after confirming that their physical condition was normal. The dose of immunogen Mel-BSA is 1 mg / kg body weight, and the immunogen and adjuvant are mixed at a ratio of 1:1. For the first immunization, the immunogen is emulsified with Freund's complete adjuvant, and for the subsequent booster immunization, it is emulsified with Freund's incomplete adjuvant Immunogen, antigen and adjuvant form a "water-in-oil" state. Take a drop of antigen and drop it on the water surface. The antigen "water-in-oil" state floats on the water surface and does not disperse, indicating that the emulsification is complete. The booster immunization was car...
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