Study On Antibody Targeting Drug Constructed By Coupled Serum Antibody Against E.coli And Gentamicin Sulfate

In recent years,The antibody treatment of cancer drug research and development has made breakthrough progress,it become a hot spot of The field of biotechnology drugs.Application of the principle of aritigen-antibody reaction Construction tumor antibody drugs,antibody and medicine combine a "biological missile",As Biological therapies of the therapy on tumor.Application of the principle of antigen-antibody reaction preparation of antibacterial drugs,antibody and antibiotics combine a "biological missile",As Biological therapies of the therapy on Bacterial Diseases,has not reported.This study coupled Serum antibody against E.coli with gentamicin sulfate1.E.coli serum antibody and polyethylene glycol and gentamicin sulfate complex PreparationTo investigate whether the E.coli antisera with gentamicin sulfate,and the combination will affect the efficacy of gentamycin sulfate and E.coli serum antibody biological activity.The first experiments using pig oil emulsion inactivated E.coli adjuvant miller,many multi-site back healthy rabbits subcutaneously,heart blood collected serum - free,using ELISA method confrontation pig serum to detect E.coli, the serum - free up to 1:2048 antibody titer.Then the combination of the two conditions for exploration to different concentrations of gentamicin sulfate,sera,PEG, a test matrix.The collection of serum antibody test gentamicin sulfate and the combination of precipitation a polyacrylamide electrophoresis,Gram staining electron microscopy.The results found that combining drugs and serum antibody good to gentamicin sulfate 800(mg):sera4(μg):PEG18(g)the combination of the largest. Thus,the serum antibody and the combination with gentamicin sulfate effectiveness and feasibility. 2.E.coli serum antibody and PEG6000 and gentamicin sulfate complex structureTo explore the combination0f structure,the experimental combination of a pair of infrared scanning and UV absorption spectrometry showed that:a combination of PEG6000 as intermediary.Serum antibody and,first of gentamicin sulfate combined with PEG6000,thus forming serum antibody - PEG6000 - gentamicin sulfate complex. The combination of hydrogen bonds.PEG6000 mainly with the amide bond of serum antibody binding.3.E.coli serum antibody and PEG6000 and gentamiein sulfate complexes targeted researchTo explore the combination of targeting the right combination of experimental indirect ELISA,fluorescence staining of electron microscopy and immunohistochemistry.ELISA results show that:the combination of both the biological activity and antibody immune function does not produce inhibition; Fluorescent antibody staining results show that:gentamicin sulfate,E.coli and PEG6000 serum antibody binding of a specific targeting of E.coli.Gentamicin sulfate, E.coli and serum antibody binding of PEG6000 mixed with the E.coli bacterium after 4 min,gentamycin sulfate,E.coli and serum antibody binding of PEG6000 combined with E.coti.Gentamicin sulfate,E.coli and serum antibody binding of PEG6000 mixed with the E.coli bacterium after 7 min,gentamycin sulfate,E.coli and serum antibody binding PEG6000 of E.coli bacteria can enter the body;Colloidal gold by immuno-electron microscopy,in the E.coli bacteria can be seen inside and outside of colloidal gold particles that combining the antibody drugs still efficient and E.coli, targeting a strong role.4.E.coil serum antibody and PEG6000 and gentamicin sulfate complex pharmacokinetics testing and evaluation of its safetyThis study also explored the combination of in vivo pharmacokinetics and safety of its detection.The results show that:Serum antibody - Gentamicin Sulfate - PEG6000 in rabbits combination of the biological half-life t1/2 to 4.83 h,medicine - AUC,area under the curve for 191.41 h.μg / mL,according to the literature reported that the sulfate - adriamycin combination of rabbits in the biological half-life of t1/21.03 h, AUC 63.42 h.μg / ml,indicating that serum antibodies - PEG6000 combination of gentamicin sulfate- pharmacokinetics and tissue distribution in a significant change in the rate of drug metabolism the slower,effective plasma concentration time,and the release of liposome similar characteristics.The serum antibody - Gentamicin Sulfate -PEG6000 with low toxicity of the mice maximal tolerance than 750 mg / kg.bw,the equivalent of 150 time the amount.Results Table serum antibody - Gentamicin Sulfate PEG6000-toxic combination of gentamicin sulfate was significantly lower than that of serum antibody - PEG6000 reduction in the role of drug toxicity.Serum antibody -Gentamicin Sulfate PEG6000-right combination of the important organs of mice had no obvious impact on the growth and development5.E.coli serum antibody and PEG6000 gentamicin sulfate complex and Pharmacodynamic StudyIn order to study the combination of pharmacodynamics,the experimental application of a combination of in vitro antibacterial tests,E.coli disease animal model for experiments.In vitro experiments showed that:Targeting gentamicin right E.coli C44103 the minimum inhibitory concentration of Gentamicin Sulfate about 1 / 10⁵.Gentamicin Sulfate targeting of Escherichiacoli_C44103minimum bactericidal gentamicin sulfate concentration is about 1 / 10³.In vitro experiments that will gentamicin sulfate and E. coli serum antibody and PEG6000 with antibacterial effect can be enhanced to reduce the dose to enhance efficacy.Animal experiments showed that the treatment model:high concentration of targeting each intramuscular gentamicin injection mL / only one drug after an efficient, the cure rates were 100%and 90%;Medium concentrations targeted by gentamicin intramuscular injection every lmL / only one with after the drug is efficient,the cure rates were 100%and 75%;Targeting low concentration intramuscular gentamicin injection each lmL / only once after treatment with 100%efficiency,the cure rate was 65%.The control group was treated with gentamicin sulfate gentamicin sulfate intramuscular injection per mL/only,after the onset of void or mice all died;Serum is ineffective treatment for the control group.Targeting gentamicin injection of E.coli in mice significantly.Clinical tests showed that:Targeting gentamicin injection can quickly kill the bacteria in pigs,reduce symptoms and improve the survival rate of piglets.Compared with gentamicin sulfate,gentamicin targeting high-dose intramuscular injection every 2.0mg/Kg,one after treatment onset piglets efficient,the cure rate was 100%;Targeting use of gentamicin dose intramuscular injection every 0.2mg/Kg,a drug after the piglets incidence rate was 100%,the cure rate is 96%;Targeting low-dose intramuscular gentamicin injection each 0.02mg/Kg two piglets after treatment morbidity rate was 96%,the cure rate was 92%;The gentamicin sulfate injection each injection 2.00mg/Kg,two times a day,3 days after the onset linked piglets efficiency of 84%,the cure rate is 76%;by targeting high-dose and gentamicin injection,the dose of gentamicin injection targeting,targeting low dose of gentamicin injection in the treatment of disease in piglets reduce speed than all of Gentamicin Sulfate Injection faster.Antisera combined with gentamicin injection can effectively kill the pathogenic E.coli,the piglets yellowscour of good clinical efficacy.Complex treatment of E.coli in chicken clinical trial showed that:anti-serum and the complex gentamicin dose administration ld,diseased disease were significantly reduced,high-dose group,2d stop dead chickens,chicken and the spirit of appetite returned;Effect of dose and high dose treatment group basic treatment consistent; low-dose group,2d stop chicken deaths,diarrhea disappeared,chicken appetite and mental state of the 3d also recovered.Gentamicin sulfate group of 2d administration began to reduce symptoms,the symptoms disappeared 4d diarrhea,loss of appetite and mental state to restore the 5d.Data from the table can be seen:anti-serum and gentamicin injection with high,medium and low doses of three pairs of chicken E.coli disease significantly,efficient and cure rates were 100%,100%,92%;100%,100%, 90%.Gentamicin sulfate group and the efficiency and the cure rate was 80%;70%.The changes in symptoms,antisera and gentamicin combination of all dose groups after administration diarrhea improve speed and the mental state of the resumption of chicken faster,and gentamicin sulfhte compared to faster improvement in the chicken diarrhea and restore chicken mental state.