Cell strain of monoclonal antibody against ciprofloxacin and monoclonal antibody thereof
A monoclonal antibody, ciprofloxacin technology, applied in the field of bioengineering, can solve the problems of high cost, long time consumption, sensitivity not meeting national standards, etc., and achieve the effects of cost saving, strong specificity and high sensitivity
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Embodiment 1
[0032] Example 1: Preparation of Immunogen—Ciprofloxacin and Bovine Serum Albumin Conjugate (CIP-BSA)
[0033] 1. Weigh 8.5g of sodium chloride, 2.85g of disodium hydrogen phosphate, 0.2g of potassium chloride, 0.27g of potassium dihydrogen phosphate, add 950mL of distilled water, add hydrochloric acid dropwise to adjust the pH to 5.0, and then add 50mL of Distilled water, configured as 1000mL phosphate buffer solution with a concentration of 0.01mol / L.
[0034] 2. Weigh 40 mg of bovine serum albumin and add 5 mL of phosphate buffer (pH 5.0, content 0.01 mol / L) to make 8 mg / mL (0.12 μmol / L) bovine serum albumin aqueous solution.
[0035] 3. Weigh 100 mg of ciprofloxacin and add 5 mL of phosphate buffer (pH 5.0, content 0.01 mol / L) to make 20 mg / mL (60.4 μmol / L) ciprofloxacin aqueous solution.
[0036] 4. Weigh 960 mg of carbodiimide and add 4 mL of phosphate buffer (pH 5.0, content 0.01 mol / L) to make an aqueous solution of 240 mg / mL (1251.9 μmol / L).
[0037] 5. Add 8mg / mL (...
Embodiment 2
[0040] Example 2: Coating source—preparation of ciprofloxacin and chicken egg serum albumin conjugate (CIP-OVA)
[0041] 1. Weigh 8.5g of sodium chloride, 2.85g of disodium hydrogen phosphate, 0.2g of potassium chloride, 0.27g of potassium dihydrogen phosphate, add 950mL of distilled water, add hydrochloric acid dropwise to adjust the pH to 5.0, and then add 50mL of Distilled water, configured as 1000mL phosphate buffer solution with a concentration of 0.01mol / L.
[0042] 2. Weigh 40 mg of chicken egg serum albumin and add 5 mL of phosphate buffer (pH 5.0, content 0.01 mol / L) to make 8 mg / mL (0.178 μmol / L) chicken egg serum albumin aqueous solution.
[0043]3. Weigh 100 mg of ciprofloxacin and add 5 mL of phosphate buffer (pH 5.0, content 0.01 mol / L) to make 20 mg / mL (60.4 μmol / L) ciprofloxacin aqueous solution.
[0044] 4. Weigh 960 mg of carbodiimide and add 4 mL of phosphate buffer (pH 5.0, content 0.01 mol / L) to make an aqueous solution of 240 mg / mL (1251.9 μmol / L).
[0...
Embodiment 3
[0048] Example 3: Preparation and purification of monoclonal antibodies
[0049] 1. Immunity
[0050] Dilute the immunogen CIP-BSA powder with phosphate buffer (pH 5.0, content 0.01mol / L) to obtain a 2 mg / mL immunogen solution, add 100 μL Freund’s complete adjuvant to 100 μL immunogen solution (2 mg / mL), After thorough mixing, 4-week-old Balb / c mice were taken and injected abdominally for immunization.
[0051] Two weeks after the immunization, take 100 μL of the immunogen solution (2 mg / mL) and add 100 μL of Freund’s incomplete adjuvant, mix thoroughly, and inject into the abdomen for immunization.
[0052] One week later, 100 μL of immunogen solution (2 mg / mL) was taken for tail vein booster immunization.
[0053] Ten days later, 50 μL of the immunogen solution (2 mg / mL) was injected into the tail vein for booster immunization.
[0054] 2. Cell Fusion
[0055] 1. Recovery and culture of myeloma cells:
[0056] 10 days before fusion, take the frozen SP2 / 0 myeloma cells o...
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