• 약학회지
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• 제59권4호
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• pp.170-176
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• 2015

In vaccine development, the major points may be induction of effective and increased levels of antibody production. This is especially the case when the antigenic sources are carbohydrates. For many years, thus, we have researched various types of formulations such as liposomal and conjugate vaccines. However, the fastidious formulation process and high costs are a problem. For this reason, there is currently a focus on utilizing immunoadjuvants. In this present study, we tested if platycodin D (PLD) from Platycodon Radix have immunoadjuvant activity against the cell wall of Candida albicans (CACW). The resulting data showed that in the murine model of antibody production, CACW combined with PLD [CACW/PLD/IFA] increased the production of antibodies specific to C. albicans when compared to the antibody production by [CACW/IFA]-induction, which was used as a negative control (P<0.05). In the case of [CACW/PLD/IFA], the antibody production was 1.4 times as that of the CFA. In addition, formulations containing either had a prolonged antibody inducing activity maintaining the initial titers of antibody as compared to the CFA formula. Cytokine profiling with the antisera displayed that the PLD produced both Th1 and Th2 immunoresponses, but Th1 dominant was much greater (P<0.05). Furthermore, [CACW/PLD/IFA] formula enhanced resistance of mice against disseminated candidiasis, whereas the CFA had no such effect. In conclusion, PLD has an immunologic activity, which is protective against the disease. Thus, PLD can be a goof candidate for a new immunoadjuvant in development of the fungal vaccine.

• 한국미생물·생명공학회지
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• 제19권3호
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• pp.272-280
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• 1991

유가식 배양에서 배지첨가속도가 하이브리도마 세포의 증식과 항체생성속도에 미치는 영향을 조사하였다. 항체 비생성속도 $q_p$값이 변화는, 회분식 배양의 경우 배양 초기에 가장 높았다가 계속 감소하는 경향을 보였으나, 배양 48시간 이후부터 배지의 첨가가 시작되는 유가식 배양에서는 qp 값이 배지 첨가직후 부터 빠른 속도로 증가하기 시작해 배양이 끝날 때까지 높은 값으로 유지되었다. 그리고 배지의 비첨 가속도 $q_f$가 증가되면서 항체의 비생성속도 $q_p$도 민감하게 증가하는 경향을 보였다. 생성된 최종 항체농도 $C_{final}$은 배지 첨가속도에 비해 큰 영향을 받지 않고 거의 비슷한 값을 보였지만 평균적으로 회분식 배양보다는 그 값이 3배 이상 증가되어 최고 220$\mu g$/ml까지 향상되었다.

• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.712-720
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• 2007

To investigate clonal variations of recombinant Chinese hamster ovary(rCHO) clones in response to culture pH and temperature, serum-free suspension cultures of two antibody-producing CHO clones(clones A and B), which were isolated from the same parental clone by the limiting dilution method, were performed in a bioreactor at pH values in the range of 6.8-7.6, and two different temperatures, $33^{\circ}C\;and\;37^{\circ}C$. In regard to cell growth, clone A and clone B displayed similar responses to temperature, although their degree of response differed. In contrast, clones A and B displayed different responses to temperature in regard to antibody production. In the case of clone A, no significant increase in maximum antibody concentration was achieved by lowering the culture temperature. The maximum antibody concentration obtained at $33^{\circ}C$(pH 7.4) and $37^{\circ}C$(pH 7.0) were $82.0{\pm}2.6$ and $73.2{\pm}4.1{\mu}g/ml$, respectively. On the other hand, in the case of clone B, an approximately 2.5-fold increase in maximum antibody concentration was achieved by lowering the culture temperature. The enhanced maximum antibody concentration of clone B at $33^{\circ}C$($132.6{\pm}14.9{\mu}g/ml$ at pH 7.2) was due to not only enhanced specific antibody productivity but also to prolonged culture longevity. At $33^{\circ}C$, the culture longevity of clone A also improved, but not as much as that of clone B. Taken together, CHO clones derived from the same parental clone displayed quite different responses to culture temperature and pH with regards antibody production, suggesting that environmental parameters such as temperature and pH should be optimized for each CHO clone.

• Journal of Applied Biological Chemistry
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• 제57권1호
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• pp.61-63
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• 2014

Aged black garlic (ABG) was extracted with 20% ethanol and water (crude extracts) and fractionated into three categories (>10, 3-10, and <3 kDa). The effect of crude extract supplements on anti-My-10 hybridoma cell growth and IgG1 antibody production was investigated in suspension culture with a chemically defined protein-free medium. We observed that supplementation of ABG to the cell culture medium stimulated anti-My-10 hybridoma cell growth and production of IgG1 antibody, particularly with fractionated ABG of low molecular weight. The stimulation depended upon the concentration and the size of the fractionated ABG. We also found that the growth-promoting activity was not correlated with high antibody production. These results suggest that fractionated ABG is a novel and promising alternative as an animal cell culture supplement.

• 약학회지
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• 제57권1호
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• pp.1-7
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• 2013

Induction of effective and increased levels of antibody production may be major points in vaccine development. This is especially the case when the antigenic sources are carbohydrates. Thus, in our Lab various types of formulations such as liposomal and conjugate vaccines have been researched. However, the fastidious formulation process and high costs are a problem. For this reason, there is currently a focus on utilizing immunoadjuvants. In this present study, we tested whether ginsenosides Re (a panaxdiol) and Rg1 (a panaxtriol) from Panax ginseng have immunoadjuvant activity against the cell wall of Candida albicans (CACW). The resulting data showed that Rd and Rg1 caused LPS-treated B lymphocytes to be proliferative. Rd had greater proliferation activity than that of Rg1. In the murine model of antibody production, CACW combined with Rd [CACW/Rd/IFA] or Rg1 [CACW/Rg1/IFA] increased the production of antibodies specific to C. albicans when compared to the antibody production by [CACW/IFA]-induction, which was used as a negative control (P<0.05). In the case of [CFA/Rd/IFA], the antibody production was almost twice as that of the CFA. In addition, formulations containing either had a prolonged antibody inducing activity as compared to the CFA formula. In conclusion, Rd and Rg1 have an immunologic activity, and yet Rd can be a better candidate than Rg1 for a new immunoadjuvant.

• 한국동물위생학회지
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• 제26권1호
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• pp.51-56
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• 2003

Avian pneumoviros(APV), also known as avian rhinotracheitis virus(ARTV), affects both turkeys and chickens and is known to be the primary causative agent of turkey rhinotracheitis (TRT). The aim of this study was to establish the presence or absence of antibodies to avian pneumovirus in the commercial poultry population of Korea. For this purpose, chicken serum samples were obtained and tested by an enzyme-linked immunosorbent assay (ELISA). The tested serum was collected in laying hens with reduction of egg production or normal in Gyeongbuk province. A total of 184 sera representing 42 different poultry farms of the Gyeongbuk region of Korea were included in this study. Laying hens of 16 different farms with reduction of egg production and laying hens of 26 different farms with clinically healthy at the time of serum sampling were considered positive to antibody against APV. In the farms with reduction of egg production, positive farm to antibody against avian pneumovirus were 11 of 16 different farms(68.8%) and positive sera were 47(58.8%) of 80 different serum. In the farms with clinically healthy flock, positive farm to antibody against avian pneumovirus were 12(46.2%) of 26 different farms and positive serum sample were 39(37.5%) of 104 different sera. According to the results tested to 42 different farms in 14 city, 8 of 14 city have flocks with antibody positive laying hens against APV, 1 of 14 city have antibody suspicious and 5 of 14 city shown antibody negative, respectively.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.293-296
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• 2002

Hyperosmotic pressure increased specific antibody productivity ($q_{Ab}$) of recombinant CHO cells (SH2-0.32) while it depressed cell growth. Thus, the use of hyperosmolar medium did not increase the maximum antibody concentration substantially. To overcome this drawback, the feasibility of biphasic culture strategy was investigated. In the biphasic culture, cells were first cultivated in the standard medium with physiological osmolality(294 mOsm/kg) for cell growth. When cells reached the late exponential phase of growth, the spent standard medium was replaced with the fresh hyperosmolar medium (522 mOsm/kg) for antibody production. The ($q_{Ab}$) in growth phase with the standard medium was 2.1 ${\mu}g/10^6cell/day$ while the ($q_{Ab}$) in antibody production phase with the hyperosmolar medium (522 mOsm/kg) was 11.1 ${\mu}g/10^6cell/day$. Northern blot analysis showed a positive relationship between the relative contenet of Ig mRNA and ($q_{Ab}$), indicating that transcriptional regulation was involved in the response of rCHO cells to hyperosmotic pressure. Due to the enhanced ($q_{Ab}$) and increased cell concentration in biphasic culture, the maximum antibody concentration obtained in biphasic culture with 522 mOsm/kg medium exchange was 161% higher than that obtained in batch culture with the standard medium. Taken together, simple biphasic culture strategy based on hyperosmotic culture for improved foreign protein production from rCHO cells is effective in improving antibody production of rCHO cells.

• 대한본초학회지
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• 제24권4호
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• pp.107-113
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• 2009

Objectives : This study was designed to investigate the effects of alcoholic fermentation beverage using pear, Bae Ro Mi In (BRMI) on airway hyperresponsiveness and immunoglobulin production in asthmatic mice Methods : We investigated the effects of BRMI on airway hyperresponsiveness by measurement of enhanced pause (Penh), and also investigated the effects on production levels of antigen specific antibody and subclasses such as IgG1, IgG2a and IgE by using ELISA methods. Prednisolone (PD, 5 mg/kg) was used as positive control. Results : Treatment with BRMI did not lowered airway hyperresponsiveness, but PD lowered significantly. Oral administration of BRMI lowered production level of ovalbumin (OVA) specific total antibody significantly. Especially, BRMI decreased IgE levels compared to non-treated control effectively. Treatment with PD lowered production levels of total antibody, IgG1 and IgE. Conclusions : These result suggest that BRMI can lower production levels of antigen specific total antibody and IgE in asthmatic mice. We also suggest that BRMI has the possibility to prevent or cure asthma through regulation of antigen specific antibody production.

• BMB Reports
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• 제42권7호
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• pp.418-420
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• 2009

We describe a method for producing polyclonal antibodies against peptide antigen cytochrome P450 1A2 and 3A4 using a tandem repeat of the epitope region and incorporation of proline residue between the repeated sequences. An ELISA assay revealed more efficient generation of polyclonal antibodies to tandem repeat peptide antigens than mono-epitope peptides. The incorporation of proline residues further stimulated antibody production.

• KSBB Journal
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• 제15권5호
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• pp.482-488
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• 2000

유전자 재조합 대장균을 이용하여 pyruvate dehydrogenase complex-E2 특이성 인간 모노클론 항체의 Fab부분이 생산되 었다. 재조합 대장균의 고밀도 배양과 이를 통한 재조합 인 간 항체의 고생산성을 확보하기 위한 최적 전략을 개발하기 위해 포도당과 초산의 영향에 대해 조사하였다. 포도당의 농 도가 높을수록 세포의 성장속도는 빨라지고 포도당의 소모속 도도 빨라짐을 알 수 있었다, 이때 포도당의 소모속도가 빨 라질수록 초산의 대사 부산물로서의 생성속도도 빨라졌다. 그러나 포도당이 고갈되면 일단 축적된 초산은 에너지원으로 소모되기 시작하고 이때의 소모속도는 배지내의 포도당의 농 도에 의존함을 알 수 있었다. 즉 초기 포도당의 농도가 어느 정도 높아 잔존 포도당의 농도가 높은 경우, 초산의 생성 속 도는 높고 에너지원으로 소모되는 속도는 느려져서 초산의 축적현상이 기하급수적으로 증가한다. 이렇게 높은 포도당 농도에 따라 축적된 초산은 항체 생산을 저해하였고, 저해를 위한 임계 초산 농도는$0.6g/\ell$이었다. 발현에 의해 항체를 생산하는 시기에서는, 포도당 농도가 높을수록 세포성장이 증대되나 초산에 의 한 저해 현상 및 catabolic repression의 영 향으로 항체 생산이 감소하였다. 따라서 발현 후 항체의 생산을 증진시키기 위해서는 포도당과 초산의 농도를 가능한 한 낮게 유지하는 것이 중요하다. 그러므로 고말도 배양과 재조합 인간 항체의 높은 생산성을 확보하기 위해서는 배양 액내의 포도당과 초산의 농도를 정밀하게 조절하는 것이 절실히 요구된다.