Ribavirin monoclonal antibody hybridoma cell strain and application thereof
A hybridoma cell line and monoclonal antibody technology, applied in biochemical equipment and methods, instruments, microorganisms, etc., can solve the problems of expensive instruments, cumbersome handling, and complicated operations, and achieve good detection sensitivity and affinity
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[0019] Embodiment 1: Preparation of hybridoma cell line 1D4
[0020] 1. Synthesis of complete antigen
[0021] Ribavirin 100mg, dissolved in 5mL of anhydrous DMF and 5mL of anhydrous pyridine mixed solution, added 50mg of succinic anhydride, reflux reaction at 60°C for 12h, and recrystallized at pH about 3, standing at 0°C to obtain white The precipitate was centrifuged and dried to obtain the ribavirin hapten. Take 10mg ribavirin hapten, add 2mL ultrapure water to dissolve, then add NHS (N-hydroxysuccinimide), EDC (carbodiimide) respectively, the molar ratio of hapten:NHS:EDC is 1: 2︰2, at room temperature, mix well, stir for 6h, then react for 12h at room temperature 25°C for activation. Take 25mg bovine serum albumin (BSA), the molar ratio of hapten and bovine serum albumin is 80:1, add 5mL0.1MpH9.6 carbonate buffer. The activated ribavirin hapten solution was slowly added dropwise to the protein solution, and reacted at room temperature for 24 hours. Dialyze with PBS b...
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