Hybridoma cell strain E secreting anti-glibenclamide monoclonal antibody and application thereof
A hybridoma cell line, monoclonal antibody technology, applied in analytical materials, biochemical equipment and methods, instruments, etc., can solve the problem of long experimental period, complex processing process, production and use of glibenclamide food additives, and import and export trade. Food hygiene monitoring work is difficult and other problems, to achieve the effect of good specificity and good detection sensitivity
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[0023] Embodiment 1 Preparation of hybridoma cell line E
[0024] (1) Synthesis of complete antigen: weigh 3.6 mg of glibenclamide (the molar ratio of glibenclamide to keyhole limpet hemocyanin (KLH) is 6000:1), 2.5 mg of N-hydroxysuccinimide (NHS) , dissolved in 300 μL N,N-dimethylformamide (DMF), stirred at room temperature for 10 minutes; then weighed 4.2 mg 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride Salt (EDC), fully dissolved in 100 μL DMF, was added to the glibenclamide solution, and stirred at room temperature for 6-8 hours (referred to as solution A). Take 6mg of KLH and dissolve it in 2mL of 0.01M borate buffer solution (BB, pH=8.6) (referred to as solution B). Then slowly add solution A to solution B drop by drop, and react overnight at room temperature; then dialyze with 0.01M PBS solution to remove unreacted small molecule hapten to obtain the complete antigen glibenclamide-KLH, and pass the ultraviolet absorption scanning method conduct identifi...
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