无针粉末注射破伤风类毒素对小鼠血清IgG抗体的影响

Effect of tetanus toxoid powder needleless injection on the concentration of serum antibody IgG in mice

本文以破伤风类毒素(tetanus toxoid,TT)为研究对象,对比自制的无针粉末注射系统与普通皮下注射方式给药对小鼠的免疫效果。考察氢氧化铝吸附破伤风类毒素(Al(OH)3-TT)粉末的制备工艺,并采用光学显微镜和激光粒度分析仪等考察其微粉学性质。结果表明,增加盐浓度、延长吸附时间均可提高吸附药量。NaCl浓度为0.4mol·L-1吸附效果较好,大于0.8mol·L-1,蛋白会发生盐析;反应10min后吸附量趋于稳定。体系pH值及温度在考察的范围内对吸附药量影响不大。根据优化后工艺制备的Al(OH)3-TT平均粒径为(60.6±4.4)μm。将自制的Al(OH)3-TT无针粉末注射剂与普通皮下注射剂对比,采用酶联免疫吸附法(ELISA)检测破伤风抗毒素IgG抗体浓度。结果表明,Al(OH)3-TT(含TT30μg)无针粉末注射小鼠,4周后破伤风抗毒素IgG抗体浓度为(6.19±0.52)U·L-1,8周后为(10.70±0.78)U·L-1,而普通皮下注射组(含TT20μg)分别为(4.25±0.58)、(7.48±0.57)U·L-1,提示自制的Al(OH)3-TT无针粉末注射剂免疫原性较好,为进一步开发破伤风类毒素无针粉末注射剂提供了技术参考。

In this study, a self-designed powder needleless injection system was compared with subcutaneous injection using a needle and syringe to deliver tetanus toxoid （TT） into mice to elicit immunity. First of all, factors influencing the prepartion of TT into powder by being absorbed on aluminium hydroxide were investigated and the micromeritic characters of Al（OH）3-TT powder were observed with optical microscope and laser particle analyzer. The results showed that salt concentration and absorption time had an enhancive effect on drug loading, but the pH value and temperature did not influence the absorption reaction obviously. The absorption reaction was optimized with sodium chloride concentration of 0.4 mol.L^-1 and lasting for 10 min. The average diameter of Al（OH）3-TT powder prepared with conditions optimized above was （60.6 ± 4.4） μm. The immunization effect of TT was determined through enzyme-linked immunosorbent assay （ELISA） of the concentration of IgG antibody elicited by TT. With delivery of Al（OH）3-TT （of 30 μg TT） by powder needleless injection to mice, the IgG antibody concentration were （6.19 ± 0.52） and （10.70 ±0.78） U.L^-1 after immunization of 4 and 8 weeks, respectively, while the values were （4.25 ±0.58） and （7.48 ±0.57） U.L^-1 by subcutaneous injection （of 20 μg TT） using a needle and syringe. The results suggested that the self-designed powderneedleless injection of Al（OH）3-TT was comparable to subcutaneous injection with a good immunity.