Development of a high sentitive oral hepatitis B vaccine using microspheres
| Project/Area Number |
09672342
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
応用薬理学・医療系薬学
|
|---|
| Research Institution | Mukogawa Women's University |
|---|
Principal Investigator |
UCHIDA Takahiro Mukogawa Women's University, Associate Professor, 薬学部, 助教授 (70203536)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MATSUYAMA Kenji Mukogawa Women's University, Professor, 薬学部, 教授 (00117251)
|
|---|
| Project Period (FY) |
1997 – 1998
|
| Project Status |
Completed (Fiscal Year 1998)
|
| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥1,500,000 (Direct Cost: ¥1,500,000)
|
|---|
| Keywords | hepatitis B Core antigen / microspheres / poly (lactide-co-glycolide) / ELISA / vaccine / 肝炎 / マイクロスフェア- |
|---|
| Research Abstract |
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.
|
Report
(3 results)
Research Products
(3 results)
