Development of a high sentitive oral hepatitis B vaccine using microspheres

使用微球开发高灵敏度口服乙型肝炎疫苗

基本信息

批准号：
09672342
负责人：
UCHIDA Takahiro
金额：
$ 1.47万
依托单位：
Mukogawa Women's University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

Purpose. To prepare poly(lactide-co-glycolide) (PLGA) microspheres containing recombinant hepatitis B core antigen (HBcAg ; Mw = 3,600,000) by a w/o/w emulsion/solvent evaporation method and evaluate the possibility of this system as a potent long-acting carrier for hepatitis B core antigen in mice.Methods. Various additives had been incorporated in the internal aqueous phase during the process of microencapsulating HBcAg, HBcAg antigenicity in the medium extracted from the prepared microspheres were measured by ELISA.Shape confirmation of the HBcAg antigen was performed by asucrose gradient velocity centrifugal technique. For in vivo study, prepared microspheres were administered subcutaneously to Balb/C mice, and the serum lgG level was determined by ELISA.Results. The inactivation of HBcAg by methylene chloride was dramatically reduced by the addition of gelatin (4-8% (w/v)) to the internal aqueous phase during the preparation. Further improvement of the loading efficiency to almost 61% resulted with cooling (4*). The prepared microspheres (4.27 mum*1.23mum) containing 0.15% HBcAg displayed burst release (50-60% within 2 days). In subcutaneous inoculation, the adjuvant effect of PLGA microspheres was almost the same as that of the complete Freund's adjuvant. Whereas oral inoculation using the microspheres was not effective.Conclusions. The pH of the added gelatin seemed to be the key to the stabilization of HBcAg from various stability tests and CD spectrum study. Finally, the possibility of using this system as a potent long-acting hepatitis B vaccine was demonostrated.

目的。通过W/O/W乳液/溶剂蒸发方法制备含有重组乙型肝炎的聚（乳酸 - 糖）（PLGA）微球（HBCAG; MW = 3,600,000），并评估该系统作为Hepatipic core antigenipeN的强大长期效果的可能性。通过ELISA测量了从制备的微球提取的培养基中的介质中，在微囊性HBCAG的过程中，已在内部水相中掺入了各种添加剂。通过ELISA测量了从准备的微球中提取的。对于体内研究，将制备的微球皮下施用到BALB/C小鼠，并由Elisa.Results确定血清LGG水平。通过在制备过程中，将明胶（4-8％（w/v））添加到内水相中，通过将明胶（4-8％（w/v））添加到甲基氯中的HBCAG大大降低。载荷效率的进一步提高到近61％，导致冷却（4*）。含有0.15％HBCAG的制备的微球（4.27 MUM*1.23MUM）显示出爆发（2天内50-60％）。在皮下接种中，PLGA微球的佐剂作用几乎与完整的Freund佐剂相同。而使用微球的口腔接种是无效的。添加明胶的pH值似乎是来自各种稳定性测试和CD光谱研究的HBCAG稳定的关键。最后，将该系统用作有效的长效丙型肝炎疫苗的可能性被解释。