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CHO 세포의 2단계 배양을 통한 Albumin-erythropoietin의 시알산 증대
임진혁(Jin-Hyuk Lim),신수아(Soo-Ah Shin),차현명(Hyun-Myoung Cha),김동일(Dong-Il Kim) 한국생물공학회 2016 KSBB Journal Vol.31 No.4
In glycoprotein, Terminal sialic acid residues of Nlinked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of CO₂ concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and 37℃ respectively and the CO₂ concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of CO₂ and 33℃ on day 5. However, the temperature and concentration of CO₂ have little effect on activity of α2,3-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.
유전자재조합 CHO 세포에서 Histone Deacetylase Inhibitor를 이용한 Albumin-erythropoietin 생산성 증진
김수진(Su-Jin Kim),서준석(Joon-Serk Seo),최성훈(Sung-Hun Choi),차현명(Hyun-Myoung Cha),임진혁(Jin-Hyuk Lim),신수아(Soo-Ah Shin),김동일(Dong-Il Kim) 한국생물공학회 2015 KSBB Journal Vol.30 No.1
Chinese hamster ovary (CHO) cells are the most widely used mammalian host for the commercial production of recombinant proteins. However, they show relatively low yields of recombinant proteins in comparison with microbial cells. Various strategies have been tried to overcome this drawback. The acetyl moieties are attached to the N-terminus of histone by histone acetyltransferase (HAT) while histone deacetylase (HDAC) removes histone-bound acetyl groups. HDAC inhibitor (HDACi), such as sodium butyrate, sodium propionate and valproic acid, can enhance specific productivity of CHO cells. Human albumin-erythropoietin (Alb-EPO) is a novel 105 kDa protein comprising recombinant human EPO fused to human albumin. In this study, we examined the effects of HDACi on the production of Alb-EPO in CHO cells with various concentrations in the range of 0-1 mM. The results showed that sodium butyrate was found to be the best HDACi for enhancing productivity. It enhanced not only the production of Alb-EPO but also the apoptosis of recombinant CHO cells.
김혜선,신수아 대한가정학회 2002 Human Ecology Research(HER) Vol.40 No.3
The purpose of this study was to develop a scale for evaluation of premarital preparation. The subjects were 412 married couples(196 males and 216 females) who had been married for 1-3 years. They were selected from Seoul, InCheon, DaeJeon, GwangJu, Pusan, and DaeGu. The questionnaire was composed of 82 items. The data was analyzed by frequencies, percentages, factor analysis, Pearson's r, and Cronbach's α. The major findings were as followings; 1. The 54 items of 82 items scale were selected through item discriminant method. The discriminant coefficients of the items(Cromer's V) ranged from .33 to .68. 2. The 7 fators that were extracted from the factor analysis explained 42% of premarital preparation. The subscales were labelled as rearing child·financial management·role division, maturity, understanding each other·love·sex, views about marriage, understanding his/her family and friends, capacity of communication and conflict resolution, and independence from parents. The criterion validity of this scale was assessed by the correlation between this premarital preparation scale and marital stability. All coefficients were significant(ranged from r=. 10 to .44, p<.01, p<.001). 3. The internal consistency of this scale was acceptable(Cronbach's α=.91).