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目的比较硫糖铝和多种糖类对抗幽门螺杆菌重组VacA卵黄抗体保护作用,为制备能耐受胃酸和蛋白酶的IgY制剂奠定基础。方法诱导DH5-αvacA-pQE30大量表达VacA蛋白,制备纯化的VacA IgY。在不同酸性的IgY液中加入同浓度各种保护剂,在相同酸性的IgY液中加入不同浓度的各种保护剂,然后加入不同浓度硫糖铝和正常胃内浓度或高浓度的胃蛋白酶,以ELISA法测定试验组IgY的剩余抗体活性(AR)。结果在pH2.0和3.0条件下,各保护剂均能提高IgY抗体活性,且硫糖铝对IgY抗体活性的保护作用优于糖类,在此条件下50%和40%硫糖铝均可完全保持IgY抗体活性。在pH2.0~3.0及正常或高胃蛋白酶浓度下,30%以上硫糖铝均可有效保护IgY的活性。结论低pH下硫糖铝对IgY抗体活性的保护作用优于糖类,且30%以上硫糖铝可增强VacA IgY对胃蛋白酶的耐受能力,是较理想的IgY保护剂。
Objective To compare the protective effects of sucralfate and various carbohydrates against the recombinant VacA yolk antibody against Helicobacter pylori, and lay a foundation for the preparation of IgY preparations that can tolerate gastric acid and protease. Methods VacA protein was induced by DH5-αvacA-pQE30 to produce purified VacA IgY. In different acidic IgY solution by adding the same concentration of various protective agents in the same acidic IgY solution by adding different concentrations of various protective agents, and then added with different concentrations sucralfate and normal gastric concentration or high concentration of pepsin, The residual antibody activity (Ig) of the test group was determined by ELISA. Results Under the conditions of pH 2.0 and 3.0, the protective agents increased the activity of IgY antibody, and sucralfate had better protective effect on the activity of IgY than 50% and 30% sucralfate Completely maintain IgY antibody activity. At pH 2.0 ~ 3.0 and normal or high pepsin concentration, more than 30% sucralfate can effectively protect IgY activity. Conclusions Sucralfate has a better protective effect on IgY activity than carbohydrate at low pH, and more than 30% sucralfate can enhance the tolerance of VacA IgY to pepsin, which is an ideal IgY protectant.