KR940007174A - Vaccine and diagnostic reagent containing Ricketer Tsutsugamu antigen - Google Patents

Vaccine and diagnostic reagent containing Ricketer Tsutsugamu antigen Download PDF

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KR940007174A
KR940007174A KR1019920016708A KR920016708A KR940007174A KR 940007174 A KR940007174 A KR 940007174A KR 1019920016708 A KR1019920016708 A KR 1019920016708A KR 920016708 A KR920016708 A KR 920016708A KR 940007174 A KR940007174 A KR 940007174A
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antigen
ricketer
tsutsugamu
tsutsugamus
eggs
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KR1019920016708A
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KR960003898B1 (en
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김태연
안창남
이지현
우규진
정세교
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허영섭
재단법인 목암생명공학연구소
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/10—Protozoa; Culture media therefor

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Abstract

본 발명에서는 세포배양에서는 대량으로 얻기 어려운 리켓챠 쯔쯔가무시 항원을 수정한 난황낭에 순화시켜 대량배양이 가능한 리켓챠 쯔쯔가무시 균주를 획득하므로써 균의 대량확보가 가능하였으며, 양이온 교환수지흡착법, 원심분리법 및 퍼콜 밀도구배 원심분리법을 병행하는 정제방법으로 높은 항원성과 면역원성을 유지하며 수율도 매우 높아 상업적으로도 가치 있는 항원을 제공할 수 있었다. 아울러, 초음파 파쇄, 길리암 및 카프의 2가지 균주의 항원의 혼합비율, 수산화알미늄 흡착 및 불활화 등의 최적조건을 결정하여 높은 감염방어 효과와 폭 넓은 항원 스펙트럼을 함께 가지고 있어, 매우 유용한 백신 및 진단시약으로서 널리 사용될 수 있었다.In the present invention, it was possible to secure a large amount of bacteria by obtaining a Rickettsia Tsutsugamu strain capable of mass culture by purifying the rickets Tsutsugamu antigen, which is difficult to obtain in large quantities in cell culture, to obtain a Ricketts Tsutsugamus strain that can be cultured in large quantities. Purification using gradient centrifugation in combination with high antigenicity and immunogenicity, yields were very high and could provide commercially valuable antigens. In addition, by determining the optimal conditions such as ultrasonic crushing, the mixing ratio of the antigens of the two strains of Giliam and Cap, aluminum hydroxide adsorption and inactivation, it has a high anti-infective effect and a broad antigen spectrum, very useful vaccine and It could be widely used as a diagnostic reagent.

Description

리켓챠 쯔쯔가무시 항원을 함유하는 백신 및 진단시약Vaccine and diagnostic reagent containing Ricketer Tsutsugamu antigen

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음Since this is an open matter, no full text was included.

제1도는 시간의 경과에 따른 리켓챠 쯔쯔가무시 길리암주 및 카프주의 항체(antibody titer) 변화를 나타내는 그래프이다.FIG. 1 is a graph showing changes in antibody titers of Rickettsia Tsutsugashimi Giliarm and Kapju over time.

Claims (15)

리켓챠 쯔쯔가무시 길리암(Rickettsia tsutsugamushi Gilliam, ATCCVR-312)주를 100대 이상 수정란에서 계대하여 대량증식이 가능해진 수정란 난황란 순화주.Ricketer Tsutsugamushi Gilliam (ATCCVR-312) Passed eggs from more than 100 fertilized eggs, which allowed mass growth. 리켓챠 쯔쯔가무시 카프(Rickettsia tsutsugamushi Karp, ATCC VR-150)주를 60대 이상 수정란에서 계대하여 대량증식이 가능해진 수정란 난황란 순화주.Ricketer Tsutsugamushi Karp (ATCC VR-150) Passage of more than 60 fertilized eggs, fertilized eggs, egg yolk eggs that are capable of mass growth. 리켓챠 쯔쯔가무시를 수정란 난황막에 증식하는 공정; 상기 수정란 난황막을 균질액으로 제조하는 공정; 상기 균질액을 불활화시키는 공정; 상기 불활화된 균질액으로 양이온 교환수지를 이용하여 난황물질을 제거하는 공정; 상기에서 얻어진 액을 원심분리하여 수용성 난황물질을 제거하는 공정; 및, 상기 수용성 난황물질이 제거된 액으로 부터 리켓챠 쯔쯔가무시 항원을 정제하는 공정을 포함하는 리켓챠 쯔쯔가무시 항원을 제조하는 방법.Ricketer Tsutsugamu process to multiply into fertilized egg yolk film; Preparing the fertilized egg yolk film as a homogeneous liquid; Inactivating the homogenate; Removing yolk material using a cation exchange resin as the inactivated homogenate; Centrifuging the liquid obtained above to remove the water-soluble yolk material; And purifying the Rickettsia Tsutsugamus antigen from the liquid from which the water-soluble egg yolk material has been removed. 제3항에 있어서, 수정란은 특이 병원이 제거된(specific pathogen free, SPF) 수정란인 방법.The method of claim 3, wherein the fertilized egg is a specific pathogen free (SPF) fertilized egg. 제3항에 있어서, 양이온 교환수지는 앰버라이트를 이용하는 방법.The method of claim 3, wherein the cation exchange resin uses amber light. 제3항에 있어서, 불활화는 포르말린으로 실행하는 방법.The method of claim 3, wherein the inactivation is performed with formalin. 제3항에 있어서, 리켓챠 쯔쯔가무시 항원의 정제는 퍼콜 밀도구배 원심분리에 의하는 방법.4. The method of claim 3, wherein the purification of Rickettsia Tsutsugamu antigen is by Percol density gradient centrifugation. 제3항의 방법에 의하여 제조된 리켓챠 쯔쯔가무시 길리암 항원.Ricketer Tsutsugashimi Gilican antigen produced by the method of claim 3. 제3항의 방법에 의하여 제조된 리켓챠 쯔쯔가무시 카프 항원.A Rickettsia Tsutsugamus Cap antigen produced by the method of claim 3. 제8항 내지 제9항의 길리암주와 카프주의 항원을 초음파 파쇄하는 공정; 상기 파쇄된 항원을 여과한 다음 적정농도로 희석하는 공정; 길리암주의 항원과 카프주의 항원을 부피비로 2:1의 비율로 혼합하는 공정; 및 상기 항원을 적정량의 수산화 알루미늄겔에 흡착시키는 공정을 포함하는 리켓챠 쯔쯔가무시 백신원액의 제조방법.Ultrasonically crushing the antigens of Gilistone and Kape of Claims 8-9; Filtering the crushed antigen and then diluting to an appropriate concentration; Mixing the antigen of Giliam and the antigen of cape in a ratio of 2: 1 by volume; And a method for adsorbing the antigen on an appropriate amount of aluminum hydroxide gel. 제10항의 방법에 의하여 제조된 백신원액을 유효성분으로 포함하는 쯔쯔가무시병 예방백신.Tsutsugamushi disease preventive vaccine comprising the vaccine stock prepared by the method of claim 10 as an active ingredient. 제8항 내지 제9항의 길리암주 및 카프주의 항원을 부피비로 1:1이 비율로 혼합하는 공정; 상기 혼합된 항원을 면양 적혈구액과 혼합하여 감작시키는 공정; 상기에서 얻어진 액으로 부터 감작되지 않은 항원을 제거하는 공정; 상기에서 얻어진 액의 적혈구 농도를 조절하는 공정; 및, 상기에서 얻어진 액을 동결건조시키는 공정을 포함하는 쯔쯔가무시병 진단용 PHA키트의 제조방법.A step of mixing the antigen of the Giliam and Kaepju of claim 8 to 9 in a ratio of 1: 1 by volume; Sensitizing the mixed antigen by mixing with sheep red blood cells; Removing the non-sensitized antigen from the liquid obtained above; Adjusting the red blood cell concentration of the liquid obtained above; And lyophilizing the solution obtained above. 제12항의 방법에 의하여 제조된 쯔쯔가무시병 진단용 PHA 키트.A PHA kit for diagnosing Tsutsugamus disease prepared by the method of claim 12. 제8항 내지 제9항의 길리암주 및 카프주의 항원을 부피비로 1:1로 혼합하는 공정; 상기 혼입된 항원을 마이크로 플레이트의 표면에 흡착시키는 공정; 상기와 같이 흡착된 항원에 완충액을 가하여 반응시키는 공정; 및, 상기 반응용액을 진공건조시키는 공정을 포함하는 쯔쯔가무시병 진단용 ELISA키트의 제조방법.A step of mixing the antigens of the Gilican and Kap strains of claims 8 to 9 in a volume ratio of 1: 1; Adsorbing the entrapped antigen to the surface of the microplate; Adding a buffer to the adsorbed antigen as described above and reacting the same; And a vacuum drying of the reaction solution. 제14항의 방법에 의해 제조된 쯔쯔가무시병 진단용 ELISA키트.ELISA kit for diagnosing Tsutsugamus disease prepared by the method of claim 14. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019920016708A 1992-09-15 1992-09-15 Preparation process of rickettsia tsutsugamushi antigen KR960003898B1 (en)

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