The Preparation And Evaluation Of E.coli Monoclonal Antibody And Amikacin Sulfate Conjugate

Using antibody-coupled drugs to target tumor cells,the antibiotics traditionally used for the treatment of E.coli disease were coupled to E.coli monoclonal antibodies via coupling agents to preserve the immunological activity of antibodies.At the same time,the drugs can be targeted to the bacteria to be aggregated,and the drug efficacy can be fully exerted.While reducing the dose of drugs,the effective dose of the drugs to the bacteria can be increased,thereby delaying the occurrence of bacterial drug resistance and effectively reducing the drug resistance.The toxic effects of the drug on the animal's body reduce drug residues.1.Preparation of E.coli Monoclonal Antibody and Amikacin Sulfate ConjugateIn this study,the positive hybridoma cells were cultured by conventional methods and screened by indirect ELISA.A large number of E.coli monoclonal antibodies were prepared in mice by animal in vivo method.The titer of ascites was more than 1:10⁵ after indirect ELISA.Then PEG6000 was used as a coupling agent to establish a square array experiment with different concentrations of E.coli monoclonal antibody,amikacin sulfate and PEG6000.The best conditions for the three combinations were explored.From the experimental results,the optimal coupling conditions were amikacin sulfate?mg?:PEG6000?g?:monoclonal antibody?mg?=800:16:6;after gram staining and electron microscopy,amikacin sulfate binds well with E.coli monoclonal antibodies.2.Pharmacokinetics and acute toxicity testing of conjugatesIn this paper,pharmacokinetic studies in rabbits and acute toxicity tests in mice were performed.The test results showed that the E.coli monoclonal antibody-PEG6000-amikacin conjugate was intramuscularly injected in rabbits and reached a peak concentration?Cmax?at0.25 h,indicating that after combining amikacin sulfate with a monoclonal antibody,Has the characteristics that can guide the drug to enter the blood quickly;the average area under the drug-time curve(AUC_0-t)is 398.039 mg/L·h,the average biological half-life(t_1/2)is 4.12h,and the half-life extension reflects After the drug is coupled to the antibody,the pharmacokinetics changes dramatically,the drug metabolism slows down,and the effective drug concentration in the blood prolongs.The maximum drug tolerance of the mouse against the conjugate exceeds 900 mg/kg.bw,which is 180 times the clinical dose.The experimental results show that the amikacin sulfate conjugate is more toxic than the amikacin sulfate drug.The language has been significantly reduced.3.In vitro antibacterial test of conjugatesThe minimal inhibitory concentration?MIC?and minimum bactericidal concentration?MBC?were determined by routine methods.The results showed that the minimum inhibitory concentration of monoclonal antibody and amikacin sulfate against E.coli was6.25?g/mL.The minimum bactericidal concentration was 6.25?g/mL,the minimum inhibitory concentration was 62.5?g/mL,the minimum bactericidal concentration was 62.5?g/mL,and the minimum inhibitory concentration of conjugate was amikacin sulfate.The1/10,minimum bactericidal concentration of the powder liquid is approximately 1/10 of that of amikacin sulfate powder.In vitro antibacterial test results showed that amikacin sulfate coupled with E.coli monoclonal antibody through PEG6000,the antibacterial effect was enhanced.