서지주요정보
Mouse cell lines 에서의 B 형 간염 바이러스 표면항원 생산과 특성에 관한 연구 = The production and characterization of hepatitis B surface antigen from mouse cell lines
서명 / 저자 Mouse cell lines 에서의 B 형 간염 바이러스 표면항원 생산과 특성에 관한 연구 = The production and characterization of hepatitis B surface antigen from mouse cell lines / 김학주.
저자명 김학주 ; Kim, Hack-Joo
발행사항 [서울 : 한국과학기술원, 1988].
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소장정보

등록번호

4105305

소장위치/청구기호

학술문화관(문화관) 보존서고

DBE 8811

SMS전송

도서상태

이용가능

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반납예정일

초록정보

For the production of hepatitis B surface antigen using the mouse cell lines, the cotransformation plasmid pKHBV-3 containing pre-S and S gene was constructed by using the hepatitis B virus DNA isolated from plasma of chronic hepatitis B carriers. As the transformants secreting hepatitis B surface antigen from mouse LTK-cell lines were isolated by hypoxanthine, aminopterin, thymidine(HAT) selection and hepatitis B surface antigen assay, hepatitis B surface antigen secreting clone(MLB5) was selected and it was confirmed secreting the 1 μg of hepatitis B surface antigen per $10^6$ cells per day. Hybridomas which are morphologically myeloma type and secrete hepatitis B surface antigen were selected by fusion between MLB5 and myeloma cell lines. KBH3 clone among the hybridomas was able to grow in mouse ascites and secreted hepatitis B surface antigen in the ascitic fluids. Also, KBH3 clone successfully secreted the hepatitis B surface antigen, while the MLB5 clone failed to produced them. The growth of KBH3 showed much faster growth kinetics than that of MLB5. Mouse monoclonal $IgG_1$ antibody to human chorionic gonadotropin from ascites fluid was purified by one-step hydroxylapatite chromatography. The yield of the purified monoclonal antibody was over 90% and essencially free of contaminating mouse IgG found in ascites fluid. Hepatitis B surface antigen in the culture media of MLB5 clone was purified by immunoaffinity chromatography using the monoclonal antibody to hepatitis B surface antigen (anti-HBs) and characterized with techniques of electron microscopy, UV-scanning spectrum and sodium dodesyl sulfate-polyacrylamide gel electrophoresis analysis. Hepatitis B surface antigen of MLB5 showed similar size, shape and same pattern of UV-spectrum such as hepatitis B surface antigen purified from hepatitis B carrier blood was shown. The polypeptides of hepatitis B surface antigen as the component of 22 nm particles were chromatographed by using the Ultrogel AcA 44 gel filtration. They showed the hepatitis B surface antigen activities and intensified the antigenicity in egg lecithin solution. A new format for agarose solid-phase enzyme immunoassay(ASEIA) has been developed in which a monoclonal anti-HBs-coupled agarose, incorporated into a 1 ml disposable plastic syringe, regulates the sample and the enzyme reaction. The sensitivity of the technique was equivalent to a commercially available enzyme immunoassay kit and was able to detect 1 μg/1 of hepatitis B surface antigen.

서지기타정보

서지기타정보
청구기호 {DBE 8811
형태사항 xviii, 128 p. : 삽도, 사진 ; 26 cm
언어 한국어
일반주기 저자명의 영문표기 : Hack-Joo Kim
지도교수의 한글표기 : 변시명
지도교수의 영문표기 : Si-Myung Byun
학위논문 학위논문(박사) - 한국과학기술원 : 생물공학과,
서지주기 참고문헌 : p. 118-125
주제 Hepatitis b virus.
Enzyme --Linked immunosorbant assay.
Affinity chromatography.
Gene expression.
유전자 발현. --과학기술용어시소러스
항원. --과학기술용어시소러스
간염 바이러스. --과학기술용어시소러스
친화성 크로마토그래피. --과학기술용어시소러스
Antigens.
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