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目的探讨酸性条件下5种糖对抗HpaA蛋黄抗体(IgY)的保护作用,为寻找提高IgY耐酸性的保护剂,制备口服防治幽门螺杆菌(H.pylori)感染的IgY制剂提供理论依据。方法建立检测抗HpaAIgY的间接ELISA法,用不同pH值的模拟胃液分别配制IgY液(IgY终浓度为1.0mg/ml),以pH1.0、2.0和3.0的模拟胃液分别配制含20%菊糖、山梨醇、葡萄糖、蔗糖、葡聚糖的IgY液(IgY终浓度为1.0mg/ml),用pH2.0的模拟胃液分别配制含不同浓度的菊糖、山梨醇的IgY液(IgY终浓度为1.0mg/ml)。上述IgY液均于37℃孵育2h,采用间接ELISA法测定IgY的抗体效价。结果 IgY的活性随着模拟胃液pH值的降低而明显下降;在酸性环境下,5种糖对IgY均有一定的保护作用,以山梨醇和菊糖的保护作用较强;在pH2.0时,40%山梨醇和30%菊糖对IgY的保护作用最强,分别能保存80.3%和73.4%的IgY活性。结论在酸性条件下,糖对IgY均有一定的保护作用,菊糖和山梨醇可作为特异性IgY的保护剂。
Objective To investigate the protective effect of five kinds of sugar against HpaA yolk antibody (IgY) under acidic conditions. In order to find a protective agent that can improve the acid resistance of IgY and provide a theoretical basis for the preparation of IgY preparations for oral prevention and treatment of H. pylori infection. Methods The indirect ELISA method was established for the detection of anti-HpaAIgY. The IgY solution (IgY final concentration was 1.0mg / ml) was prepared by simulating gastric juice with different pH values. The simulated gastric fluid with pH1.0, 2.0 and 3.0 was used to prepare 20% , IgY (sorbitol, glucose, sucrose, dextran, IgY, and IgY at a final concentration of 1.0 mg / ml) were prepared. IgY solution containing different concentrations of inulin and sorbitol 1.0 mg / ml). The IgY solution were incubated at 37 ℃ 2h, indirect ELISA method to determine IgY antibody titer. Results The activity of IgY decreased significantly with the decrease of pH of gastric juice. Under acidic conditions, all the five kinds of sugars had some protective effects on IgY and strong protective effect with sorbitol and inulin. At pH 2.0, 40% sorbitol and 30% inulin have the strongest protective effect on IgY, which can save 80.3% and 73.4% IgY activity, respectively. Conclusion Under acidic conditions, the sugar has a certain protective effect on IgY, inulin and sorbitol can be used as specific IgY protective agent.