초록

본 연구에서는 바이러스 유사입자를 기반으로 한 재조합 노다바이러스 주사용 및 경구용 백신을 개발하였고 생산된 노다바이러스 백신 시제품을 양식 현장 어체에 적용하여 백신 효능을 입증하였음.

(출처 : 요약서 3p)

Abstract ▼

Purpose & Contents
Purpose
1. Development of recombinant nodavirus vaccine based on virus-like particle (VLP)
2. Production of trial product of recombinant nodavirus vaccine as an injectable vaccine, which is less expensive and highly efficient, and field trial in the fish cultivation
3.

Purpose & Contents
Purpose
1. Development of recombinant nodavirus vaccine based on virus-like particle (VLP)
2. Production of trial product of recombinant nodavirus vaccine as an injectable vaccine, which is less expensive and highly efficient, and field trial in the fish cultivation
3. Production of trial product of recombinant yeasts expressing nodavirus capsid protein as an oral vaccine, which is less expensive and highly efficient, and field trial of the trial product in the fish cultivation

Contents
1. Establishment of highly efficient production condition for recombinant nodavirus vaccine antigen and production condition for maximizing immunogenicity of the vaccine antigen.
2. Establishments of standard operating procedures for large-scale productions of recombinant nodavirus vaccines.
3. Verification of stability and safety of recombinant nodavirus vaccine.
4. Efficacy test against recombinant nodavirus vaccine (including field trial in fish cultivation).

Results
1. Development of cell lines for producing vaccine antigens (oral and injectable vaccine)
- Yeast cell lines for producing vaccine antigens were established.
- Expression stabilities of the antigens in the yeast cell lines when the passage number increased were confirmed.

2. Establishments of systems for large scale yeast fermentation (oral and injectable vaccines)
- Culture condition of yeast cells which is potentially usable for large scale fermentation was established.
- Condition for purifying the antigens for injectable vaccine was established, and the content was published as article and patent.

3. Establishments of fish systems to investigate the vaccine efficacy (oral and injectable vaccine).
- The vaccine efficacies were verified in convict grouper and olive flounder model.
- In the convict grouper model, the vaccine efficacies were verified as presences of serum antibodies against nodavirus capsid protein following vaccination, and survival rates post nodavirus challenge to the vaccinated fish were checked.
- In olive flounder model, the vaccine efficacies were verified as generations of the serum antibodies against nodavirus capsid protein.

4. Verifications of stabilities and safeties of injectable and oral vaccines
- The stabilities of vaccine antigens were verified in various conditions (the condition for preserving antigen was varied and the vaccine antigens were exposed to unfavorable conditions), and optimum conditions for preserving the antigen’s properties were established. Corresponding contents were published as article.

5. Establishments of standard operating procedures for producing injectable and oral vaccines
- The standard operating procedures for vaccine productions were established, and stabilities in antigen productions were confirmed from three production batches when it was conducted according to the operating procedures.

6. Suggestion of parameters for evaluating vaccination programs (injectable and oral vaccines)
- The guidelines for evaluating antibody generations following vaccination program were established.

7. Development of optimized formulation for oral vaccine
- Vaccine formulation to increase voluntary consumption of fish were established.

8. Productions of prototype vaccines (oral and injectable vaccines)
- The prototype vaccines were manufactured, and their abilities to elicit antibodies were evaluated.

9. Verifications of efficacies of injectable and oral vaccines (field trials to fish hatchery)
- The generations of antibodies and protective immunities against nodavirus following vaccinations were verified in convict grouper. Corresponding contents were published in article (SCI; top 10% of journal)

10. Studies for improving manufacturing processes which reduce production costs of injectable and oral vaccines were conducted.
- The cell culture condition for reducing use of galactose was selected. It was confirmed that the changed culture condition satisfies the yield of vaccine antigen we aimed.

11. Assessment of sensitivity of olive flounder to nodavirus infection
- The pathogenicity of nodavirus to olive flounder was investigated to assess the applicability of the olive flounder model as verifying the vaccine efficacy.

12. Investigation of immune response of vaccine following use of adjuvant
- The adjuvant was applied to reduce the amount of vaccine antigen, and the antibody responses as a function of the adjuvant were investigated.

Expected Contribution
1. Improvement of constitution in domestic fish cultivation area and enhancement of farm fish yield will be possible through providing nodavirus vaccine.
2. Aquisition of international competitiveness to export farm fish will be possible.
3. Aquisition of technical superiority to control viral disease of fish in international research field through development of recombinant virus vaccine for fish will be possible.

(출처 : SUMMARY 6p)

목차 Contents

• 표지 ... 1제출문 ... 2보고서 요약서 ... 3요약문 ... 4SUMMARY ... 6Index ... 8목차 ... 8제1장. 연구개발과제의 개요 ... 9 1. 연구개발 목적 ... 9 2. 연구개발의 필요성 ... 10 3. 연구개발 범위 ... 18제2장. 국내외 기술 개발 현황 ... 22 1. 국외 기술, 산업동향 ... 22 2. 국내 기술, 산업동향 ... 33 3. 특허 동향 ... 38제3장. 연구 수행 내용 및 성과 ... 56 1. 백신 효능 검증을 위한 어류 시스템 최적화 ... 58 2. 주사용/경구용 백신의 대량 생산 공정 확립 ... 70 3. 주사용/경구용 백신 생산 세포주 구축 및 항원 특성화 ... 76 4. 주사용/경구용 백신 생산에 관한 공정서 수립 ... 80 5. 주사용/경구용 백신의 안정성(stability) 확인 ... 86 6. 주사용/경구용 백신의 안전성(safety) 확인 ... 90 7. 경구용 백신의 제형 최적화 ... 97 8. 주사용/경구용 백신의 실험실 수준 효능 확인 ... 98 9. 주사용/경구용 백신의 양식 현장 효능 확인(1차 실험) ... 104 10. VLP 백신 항원의 질(quality)에 관한 기준 마련 ... 112 11. 백신접종 프로그램의 지표 제시 ... 115 12. 주사용/경구용 백신 시제품 제작 및 면역원성 확인 ... 117 13. 주사용/경구용 백신의 양식 현장 효능 확인(2차 실험) ... 123 14. 주사용 백신의 면역보조제 적용에 따른 면역 반응 확인 ... 130 15. 백신 동결 건조 제형에 대한 항원 안정성 시험 ... 133 16. 백신의 생산 단가 감소를 위한 공정 개선 연구 ... 138 17. 넙치의 노다바이러스에 대한 감염 감수성 시험 ... 144 18. 백신의 경제성 평가 ... 146제4장. 목표 달성도 및 관련 분야 기여도 ... 150제5장. 연구개발성과의 활용계획 ... 156제6장. 연구 과정에서 수집한 해외 과학기술 정보 ... 156제7장. 연구개발성과의 보안등급 ... 158제8장. 국가과학기술종합정보시스템에 등록한 연구시설·장비현황 ... 158제9장. 연구개발과제 수행에 따른 연구실 등의 안전 조치 이행 실적 ... 158제10장. 연구개발과제의 대표적 연구 실적 ... 158제11장. 기타 사항 ... 159제12장. 참고 문헌 ... 160끝페이지 ... 162