點帶石斑虹彩病毒次單位口服疫苗及包覆技術之研發

Abstract

本論文使用微膠囊包覆技術(Microencapsulation)包覆點帶石斑虹彩病毒(orange spotted grouper iridovirus, OSGIV)次單位疫苗，使用的包覆材質是褐藻酸鈉，具有無毒、不影響抗原的免疫性及低成本的優勢，以褐藻酸鈉當作包覆材質製成的微膠囊(Microcapsule)在酸性環境中相當穩定，不會被強酸崩解而釋出內含物。相對地，微膠囊在鹼性環境中會因為pH值改變而慢慢崩解，因此，微膠囊可以保護疫苗通過胃酸環境，抵達腸道，使疫苗在弱鹼環境的腸道內釋放。本論文使用幾丁聚醣作為添加物，增加微膠囊的蛋白質乘載率及包覆率，依據本論文的微膠囊製程法，蛋白質乘載率為7.74%±1.23%，包覆率為50.46%±8.03%。 將微膠囊以外添加的方式與魚用飼料混合，連續餵食4週。腸道內專一性抗體出現顯著差異的時間比血清中專一性抗體早。每週持續追蹤血清中專一性抗體變化，結果顯示，口服低劑量組的血清內專一性抗體足夠維持8週，口服高劑量組的血清內專一性抗體則維持10週。根據本論文研究的微膠囊製程法顯示，包覆點帶石斑虹彩病毒之次單位口服疫苗，可以成功將疫苗送至後腸並引發免疫反應，產生專一性抗體。

The vacccine has to be protected against gastric acid and digestive system. My thesis uses microencapsulation technique to encapsulate OSGIV (orange spotted grouper iridovirus, OSGIV) subunit vaccine from digestive system. Microencapsulation material is sodium alginate, its properties are low toxic, low antigenic and low cost. Using sodium alginate as microencapsulation material is called microcapsule. It is acid-stable ,but it will be collapsed when pH value arises in base environment. Therefore, microcapsule can make vaccine safely passing through stomach and be released in postgut. In my thesis, microcapsule loading rate is 7.74%±1.23%，encapsulation efficiency is 50.46%±8.03%. Mix microcapsule and grouper artificial feed and keep feeding Epinephelus lanceolatus for four weeks, I find out that the groups of fed oral vaccine’s specific antibody in gut arise faster then serum. The results are the group of fed low dosage oral vaccine’s specific antibody can keep eight weeks and the group of fed high dosage oral vaccine’s specific antibody can keep ten weeks. Based on my thesis, the microcapsule of oral vaccine can induce immune response to produce specific antibody successfully.