서지주요정보
하이브리도마 세포의 고농도 연속배양을 통한 단일 클론 항체의 생산 = Continuous cultures of high density hybridoma cells for the production of monoclonal antibodies
서명 / 저자 하이브리도마 세포의 고농도 연속배양을 통한 단일 클론 항체의 생산 = Continuous cultures of high density hybridoma cells for the production of monoclonal antibodies / 오덕재.
저자명 오덕재 ; Oh, Duk-Jae
발행사항 [대전 : 한국과학기술원, 1992].
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등록번호

8002472

소장위치/청구기호

학술문화관(문화관) 보존서고

DCE 92015

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초록정보

Monoclonal antibodies (Mabs) have been widely used in the diagnostics, separation and purification of proteins since their discovery in 1975 and recent application has been extended to therapeutic areas. Mabs are commercially produced by using homogeneous suspension cultures or immobilized cell cultures. But commercial culture systems may cause some problems in large scale applications. In this study, continuous cultures of high density hybridoma cells secreting Mabs against hCG were investigated for the production of Mabs. In suspension cultures, culture characteristics such as specific rates of cell growth, Mab production, and glucose consumption were studied. In the case of hybridoma Alps 25-3, 0.065 $h^{-1}$, 1.0μg Mab/$10^6$ cells·h, and 0.1 mg glucose/$10^6$ cells·h were obtained, respectively. In comparison with batch or continuous suspension cultures, two or three times higher cell densities and Mab concentrations were obtained by using cell settler. But long term continuous culture failed, because the media conditions within cell settler got out of control. In membrane cell recycle system, fragility of hybridoma led to cell death by disruption of cellular membrane, when they were recirculated by pumping systems. For successful operations, mild pumping system should be developed to recirculate suspended cells. In immobilized cell culture systems, cells were immobilized and cultivated within dual hollow fiber, Ca-alginate capsule and cylindrical depth filter. In dual hollow fiber bioreactor (DHFBR), hybridomas could be successfully cultured in high density of ca. $1.87\times 10^8$ cells/mL for two months. The Mab productivity reached to 205 mg/L day. The mathematical analysis of oxygen transfer in DHFBR showed that oxygen transfer to immobilized cell matrix was improved much compared with conventional hollow fiber bioreactor. Encapsulation of hybridomas using only Ca-alginate gel seemed to be simpler and more economical than the Lim's encapsulation process. High density cells of $1.5\times 10^8$ cells/mL in capsules resulted in high Mab productivity of 652.8 mg/L day. But long term continuous culture could not be carried out in Ca-alginate capsule system because supply of nutrients and removal of wastes were limited as grown cells packed the entire intra-capsule space, that was common to encapsulation systems. The analysis of oxygen transfer in the Ca-alginate capsules showed that oxygen transfer was seriously limited. A newly developed depth filter perfusion system (DFPS) based on the immobilization of hybridoma cells within cylindrical depth filter matrix yielded a high Mab productivity of 774 mg/L day. The cells could be easily immobilized, and high density continuous culture could be successfully carried out in DFPS. In the study of growth regulation of hybridomas, the cell growth was retarded by using growth inhibitors (hydroxy urea and thymidine), but culture time of hybridomas would not be elongated in batch stationary cultures. Inoculum cell density was found to be an important parameter in determining the viability of hybridomas cultivated in serum-deficient medium.

서지기타정보

서지기타정보
청구기호 {DCE 92015
형태사항 xiv, 163 p. : 삽도 ; 26 cm
언어 한국어
일반주기 저자명의 영문표기 : Duk-Jae Oh
지도교수의 한글표기 : 장호남
지도교수의 영문표기 : Ho-Nam Chang
학위논문 학위논문(박사) - 한국과학기술원 : 화학공학과,
서지주기 참고문헌 : p. 153-163
주제 Cell culture.
Membrane reactors.
단일클론성 항체. --과학기술용어시소러스
연속 배양. --과학기술용어시소러스
Monoclonal antibodies.
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