11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and application thereof
A hybridoma cell line and monoclonal antibody technology, applied in the field of immunoassay, can solve the problems of high cost, time-consuming, cumbersome steps, etc.
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[0060] A hybridoma cell line that secretes 11-alpha hydroxyprogesterone monoclonal antibody and a preparation method thereof, specifically comprising the following steps:
[0061] (1) Preparation of 11-α hydroxyprogesterone complete antigen:
[0062] a. The derivation route of 11-α hydroxyprogesterone hapten is as follows:
[0063]
[0064] Weigh 10mg of 11-α-hydroxyprogesterone, dissolve it in 0.6mL methanol:water:pyridine (4:1:1) solution, add 4mg carboxymethylhydroxylamine hemihydrochloride (CMO), stir in a water bath at 65°C 2h, let stand overnight at room temperature in the dark. Subsequently, the solution was blown dry by nitrogen, resuspended in 1 mL of chloroform, and extracted three times with ultrapure water. The organic phase was collected, dried with nitrogen again, and the final product was dissolved in 1 mL of anhydrous N,N-dimethylformamide (DMF), namely the 11-α hydroxyprogesterone hapten.
[0065] b. Weigh 7.5 mg of hapten and dissolve it in 500 μL DMF, ...
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