{"id":{"repo_id":"ajou","oai_identifier":"oai:repository.ajou.ac.kr:201003/10851"},"canonical_url":"https://search.dev.ndltd.org/etd/ajou/oai:repository.ajou.ac.kr:201003/10851","repository":{"repo_id":"ajou","name":"Ajou University","base_url":"http://repository.ajou.ac.kr/oai/request"},"display":{"title":"Generation of chickenized 3D8 scFv antibody from mouse monoclonal anti-DNA antibody by CDRs-grafting","abstract":"3D8 single chain variable fragment (scFv) derived from a MRL-lpr/lpr mouse is a catalyric anti nucleic acid antibody that has ability of cellular internalization and DNA/RNA-hydrolyzing activity. It has been reported that cells treated with 3D8 scFv protein showed the down-regulation against replication of RNA-genomed viruses and this anti-viral activity should be due to the cellular internalizaing and DNA/RNA- hydrolyzing activities of 3D8 scFv. To extend the possible applications of anti-viral activity of 3D8 scFv, we have developed a chickenized antibody of 3D8 scFv by CDRs (complementarity-derermining regions) grafting. A chickenized 3D8 scFv variant (CK) was generated by transferring six CDRs of 3D8 scFv between FRs (framework regions) of a chicken antibody which reveals high solubility and expression level in bacterial culture. CK6 of chickenized 3D8 scFv variant (CK1~7) retained the DNA-binding, DNA-hydrolysis, and cellular internalizing activities similar to that of the original mouse 3D8 scFv (m3D8). Our results demonstrate that chckenized 3D8 scFv can be generated by grafting CDRs.","abstract_html":"3D8 single chain variable fragment (scFv) derived from a MRL-lpr/lpr mouse is a catalyric anti nucleic acid antibody that has ability of cellular internalization and DNA/RNA-hydrolyzing activity. It has been reported that cells treated with 3D8 scFv protein showed the down-regulation against replication of RNA-genomed viruses and this anti-viral activity should be due to the cellular internalizaing and DNA/RNA- hydrolyzing activities of 3D8 scFv. To extend the possible applications of anti-viral activity of 3D8 scFv, we have developed a chickenized antibody of 3D8 scFv by CDRs (complementarity-derermining regions) grafting. A chickenized 3D8 scFv variant (CK) was generated by transferring six CDRs of 3D8 scFv between FRs (framework regions) of a chicken antibody which reveals high solubility and expression level in bacterial culture. CK6 of chickenized 3D8 scFv variant (CK1~7) retained the DNA-binding, DNA-hydrolysis, and cellular internalizing activities similar to that of the original mouse 3D8 scFv (m3D8). Our results demonstrate that chckenized 3D8 scFv can be generated by grafting CDRs.","abstract_has_math":false,"creators":["노, 주호"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["권, 명희","대학원 의생명과학과","201224127"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-11-10T01:32:10Z","date_published":"2014-11-10T01:32:10Z","updated_at":"2026-07-24T00:51:44Z","subjects":["마우스 3D8 scFv","항-핵산 항체","CDR-이식","닭화 (chickenized)","3D8 scFv","anti nucleic acid antibody","DNA/RNA hydrolyzing activity","chickenization","CDR-grafting","Immunogenicity"],"languages":["ko"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000016301","000000016301"],"render_values":[{"text":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000016301","href":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000016301","code":true},{"text":"000000016301","href":null,"code":true}]}]},"links":{"outbound_url":"http://repository.ajou.ac.kr/handle/201003/10851","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["권, 명희","대학원 의생명과학과","201224127","노, 주호"]},{"key":"dc:creator","label":"Author","values":["노, 주호"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2014-11-10T01:32:10Z","2014"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","Theses"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["마우스 3D8 scFv","항-핵산 항체","CDR-이식","닭화 (chickenized)","3D8 scFv","anti nucleic acid antibody","DNA/RNA hydrolyzing activity","chickenization","CDR-grafting","Immunogenicity"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["ko"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://repository.ajou.ac.kr/handle/201003/10851","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000016301","000000016301"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["3D8 single chain variable fragment (scFv) derived from a MRL-lpr/lpr mouse is a catalyric anti nucleic acid antibody that has ability of cellular internalization and DNA/RNA-hydrolyzing activity. It has been reported that cells treated with 3D8 scFv protein showed the down-regulation against replication of RNA-genomed viruses and this anti-viral activity should be due to the cellular internalizaing and DNA/RNA- hydrolyzing activities of 3D8 scFv. To extend the possible applications of anti-viral activity of 3D8 scFv, we have developed a chickenized antibody of 3D8 scFv by CDRs (complementarity-derermining regions) grafting. A chickenized 3D8 scFv variant (CK) was generated by transferring six CDRs of 3D8 scFv between FRs (framework regions) of a chicken antibody which reveals high solubility and expression level in bacterial culture. CK6 of chickenized 3D8 scFv variant (CK1~7) retained the DNA-binding, DNA-hydrolysis, and cellular internalizing activities similar to that of the original mouse 3D8 scFv (m3D8). Our results demonstrate that chckenized 3D8 scFv can be generated by grafting CDRs.","자가면역 마우스 (MRL-lpr/lpr)에서 유래한 3D8 단일가닥 가변부위(single-chain variable fragment: scFv)는 세포 내 침투능력과 DNA/RNA 가수분해능력을 가진 촉매성 항-핵산 항체이다. 이전 연구에서 마우스 3D8 scFv를 세포에 처리하고 RNA-게놈 바이러스를 감염시켰을 때 바이러스의 복제가 억제되는 것이 관찰 되었다. 이러한 항-바이러스 활성은 마우스 3D8 scFv의 가수분해능력 때문으로 알려져 있다. 마우스 3D8 scFv의 항-바이러스 활성을 닭에게 적용하기 위해 본 연구에서는 CDR(complementarity-determining regions)-이식 전략을 이용하여 닭화 3D8 scFv 항체(chickenized 3D8 scFv antibody)를 제작하고자하였다. 이를 위하여 수용성이 높다고 알려진 닭 항체 뼈대 (Framework regions, FRs)에 마우스 3D8 scFv의 6개 CDRs을 옮김으로써 7가지 닭화 3D8 scFv(CK1-7)를 구축하였다. 이들 닭화 3D8 scFv 단백질을 대장균에서 발현 및 정제하여, 기존 마우스 3D8 scFv의 DNA-결합능력과 DNA-가수분해능력 및 세포 내 침투능력을 유지하는지 조사하였다. 그 결과 닭화 3D8 scFv항체 중 하나인 CK6가 마우스 3D8 scFv의 모든 활성을 동일하게 유지하는 것을 관찰하였다. 본 연구는 3D8의 본래 활성을 그대로 유지하면서 닭에게 적용할 수 있는 새로운 닭화 항체 제조의 가능성을 제시한다.","국문요약ⅰ 차례ⅱ 그림 차례ⅳ 표 차례ⅵ Ⅰ. 서론1 Ⅱ. 재료 및 방법6 A. 대장균에서 닭화 3D8 scFv 항체 발현 및 정제6 B. 전기영동법 (SDS-PAGE) 시행 C. 면역효소흡착법 (ELISA)7 D. 공초점 현미경 분석7 E. 유세포 분석(flow cytrometry) F. FRET-based DNA-hydrolysis assay9 G. 면역원성 (immunogenicity)조사 1. m3D8 scFv와 닭화 3D8 scFv의 면역 및 채혈9 2. protein A tag을 제거한 3D8 scFv와 CK6 정제10 3. 혈청에서 면역원성 분석을 위한 ELISA11 Ⅲ. 결과13 A. 닭화 3D8 scFv 발현벡터 제작13 B. 박테리아에서 닭화 3D8 scFv 항체의 발현 및 정제16 C. 닭화 3D8 scFv의 DNA 결합 능력 확인18 D. 닭화 3D8 scFv의 세포 내 유입능력20 E. 닭화 3D8 scFv의 핵산가수분해 능력23 F. 닭화 3D8 scFv의 특징 요약23 G. 닭화 3D8 scFv의 면역원성26 Ⅳ. 고찰32 Ⅴ. 결론34 참고문헌35 Abstract40","Master"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Generation of chickenized 3D8 scFv antibody from mouse monoclonal anti-DNA antibody by CDRs-grafting","마우스 유래 항-DNA 단클론 항체로 부터 CDR-이식방법을 통한 닭화 3D8 scFv 항체 제조"]}]}],"canonical_facts":{"dc:contributor":["권, 명희","대학원 의생명과학과","201224127","노, 주호"],"dc:creator":["노, 주호"],"dc:date":["2014-11-10T01:32:10Z","2014"],"dc:description":["3D8 single chain variable fragment (scFv) derived from a MRL-lpr/lpr mouse is a catalyric anti nucleic acid antibody that has ability of cellular internalization and DNA/RNA-hydrolyzing activity. It has been reported that cells treated with 3D8 scFv protein showed the down-regulation against replication of RNA-genomed viruses and this anti-viral activity should be due to the cellular internalizaing and DNA/RNA- hydrolyzing activities of 3D8 scFv. To extend the possible applications of anti-viral activity of 3D8 scFv, we have developed a chickenized antibody of 3D8 scFv by CDRs (complementarity-derermining regions) grafting. A chickenized 3D8 scFv variant (CK) was generated by transferring six CDRs of 3D8 scFv between FRs (framework regions) of a chicken antibody which reveals high solubility and expression level in bacterial culture. CK6 of chickenized 3D8 scFv variant (CK1~7) retained the DNA-binding, DNA-hydrolysis, and cellular internalizing activities similar to that of the original mouse 3D8 scFv (m3D8). Our results demonstrate that chckenized 3D8 scFv can be generated by grafting CDRs.","자가면역 마우스 (MRL-lpr/lpr)에서 유래한 3D8 단일가닥 가변부위(single-chain variable fragment: scFv)는 세포 내 침투능력과 DNA/RNA 가수분해능력을 가진 촉매성 항-핵산 항체이다. 이전 연구에서 마우스 3D8 scFv를 세포에 처리하고 RNA-게놈 바이러스를 감염시켰을 때 바이러스의 복제가 억제되는 것이 관찰 되었다. 이러한 항-바이러스 활성은 마우스 3D8 scFv의 가수분해능력 때문으로 알려져 있다. 마우스 3D8 scFv의 항-바이러스 활성을 닭에게 적용하기 위해 본 연구에서는 CDR(complementarity-determining regions)-이식 전략을 이용하여 닭화 3D8 scFv 항체(chickenized 3D8 scFv antibody)를 제작하고자하였다. 이를 위하여 수용성이 높다고 알려진 닭 항체 뼈대 (Framework regions, FRs)에 마우스 3D8 scFv의 6개 CDRs을 옮김으로써 7가지 닭화 3D8 scFv(CK1-7)를 구축하였다. 이들 닭화 3D8 scFv 단백질을 대장균에서 발현 및 정제하여, 기존 마우스 3D8 scFv의 DNA-결합능력과 DNA-가수분해능력 및 세포 내 침투능력을 유지하는지 조사하였다. 그 결과 닭화 3D8 scFv항체 중 하나인 CK6가 마우스 3D8 scFv의 모든 활성을 동일하게 유지하는 것을 관찰하였다. 본 연구는 3D8의 본래 활성을 그대로 유지하면서 닭에게 적용할 수 있는 새로운 닭화 항체 제조의 가능성을 제시한다.","국문요약ⅰ 차례ⅱ 그림 차례ⅳ 표 차례ⅵ Ⅰ. 서론1 Ⅱ. 재료 및 방법6 A. 대장균에서 닭화 3D8 scFv 항체 발현 및 정제6 B. 전기영동법 (SDS-PAGE) 시행 C. 면역효소흡착법 (ELISA)7 D. 공초점 현미경 분석7 E. 유세포 분석(flow cytrometry) F. FRET-based DNA-hydrolysis assay9 G. 면역원성 (immunogenicity)조사 1. m3D8 scFv와 닭화 3D8 scFv의 면역 및 채혈9 2. protein A tag을 제거한 3D8 scFv와 CK6 정제10 3. 혈청에서 면역원성 분석을 위한 ELISA11 Ⅲ. 결과13 A. 닭화 3D8 scFv 발현벡터 제작13 B. 박테리아에서 닭화 3D8 scFv 항체의 발현 및 정제16 C. 닭화 3D8 scFv의 DNA 결합 능력 확인18 D. 닭화 3D8 scFv의 세포 내 유입능력20 E. 닭화 3D8 scFv의 핵산가수분해 능력23 F. 닭화 3D8 scFv의 특징 요약23 G. 닭화 3D8 scFv의 면역원성26 Ⅳ. 고찰32 Ⅴ. 결론34 참고문헌35 Abstract40","Master"],"dc:format":["application/pdf"],"dc:identifier":["http://repository.ajou.ac.kr/handle/201003/10851","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000016301","000000016301"],"dc:language":["ko"],"dc:subject":["마우스 3D8 scFv","항-핵산 항체","CDR-이식","닭화 (chickenized)","3D8 scFv","anti nucleic acid antibody","DNA/RNA hydrolyzing activity","chickenization","CDR-grafting","Immunogenicity"],"dc:title":["Generation of chickenized 3D8 scFv antibody from mouse monoclonal anti-DNA antibody by CDRs-grafting","마우스 유래 항-DNA 단클론 항체로 부터 CDR-이식방법을 통한 닭화 3D8 scFv 항체 제조"],"dc:type":["Thesis","Theses"]},"updated_at":"2026-07-24T00:51:44Z"}