• KSBB Journal
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• 제17권4호
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• pp.326-332
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• 2002

크로마토그래피 공정 성능 검증의 한 항목인 담체의 수명 검증을 위해 자동화된 미니 크로마토그래피 시스템(MiniValChrom)을 개발하였다. 이 시스템은 크로마토그래피공정의 자동화된 반복수행, 크로마토그래피 각 step의 작동순서 및 공정조건의 자유로운 구성, on-line 실시간 모니터링 및 제어, 여러 담체 수명 검증법의 method file 저장기능 등을 갖추었다. MiniValChrom을 사용하여 BSA와 Cibacron Blue 3G-A를 각각 모델 단백질과 담체로 담체 수명 검증실험을 사례연구로 수행하였다. 담체 수명의 감소는 크로마토그래피 공정을 반복수행하면서 5 cycle 마다 변화하는 HETP값을 측정하여 HETP 값이 1 cm 이상 될 때까지 반복 수행함으로써 담체 수명을 결정할 수 있었다. 본 연구를 통해 개발된 MiniValChrom은 다른 검증 항목이나 multi-product 생산공장 내 크로마토그래피 공정 검증에도 유용하게 쓰일 수 있을 것으로 기대된다.

• KSBB Journal
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• 제22권4호
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• pp.213-217
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• 2007

첨가제가 단백질 약물 방출 속도 및 약물 제제 제조 및 구조에 미치는 영향을 고찰하였다. 친수성 첨가제인 D-sorbitol의 경우 친유성 첨가제보다 단백질 약물 방출 속도를 감소시킬 수 있었으며 최적의 농도는 3% (w/v)로 나타났다. 또한 제제 제조시 점도를 낮게 유지할 뿐 아니라 상분리 없는 균일한 pluronic 용액상태를 유지하여 약물이 첨가될 경우에 균일한 약물제제를 만들 수 있었다. 한편 D-sorbitol은 pluronic 수용액의 CMC를 낮추고 마이셀 표면에 작용하여 구조를 강화하는 역할을 수행하는 것으로 보인다. 따라서 pluronic 제제에 D-sorbitol을 첨가하여 단백질 약물의 안정성을 향상시키고 효과적인 약물전달 시스템을 설계할 수 있었다.

• 폴리머
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• 제26권4호
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• pp.415-421
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• 2002

E-beam 전조사법을 이용하여 HPP-g-styrene 공중합체와 술폰화 반응을 통한 술폰화 HPP-g-styrene 섬유이온교환체를 합성하였다. 그라프트율은 스티렌 단량체 농도가 증가함에 따라 증가하였으며 스티렌 단량체 농도가 80%에서 그라프트율이 128%로 최대를 나타냈다. 술폰화율은 그라프트율이 증가함에 따라 증가하는 경향을 나타내었으며, 그라프트율이 100%일 때 13.4%로 최대값을 나타내었다. 술폰화 HPP-g-styrene 섬유이온교환체의 이온교환용량은 약 3.42 meq/g으로써 흡착 성능이 매우 우수한 소재임을 확인하였다. BET 분석결과 술폰화 HPP-g-styrene의 비표면적은 62.54 $m^2/g$, 기공크기는 25 $\AA$으로 반응전보다 비표면적은 감소하였고 기공크기는 약간 증가하는 경향을 보였다. 또한 Bovine Serum Albumin (BSA) 흡착 실험 결과 술폰화도가 증가함에 따라 BSA 흡착 용량이 증가하는 경향을 나타내었으며, 술폰화도 13.4%에서 BSA 흡착용량 3.8 mg/g으로 최대를 나타내었다. 따라서 본 연구에서 합성한 섬유이온교환체가 BSA 흡착.분리에 적합한 소재임을 확인하였다.

• 약학회지
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• 제47권3호
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• pp.167-175
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• 2003

Eleutherococcus senticosus is a typical oriental folk medicinal herb. It has been used clinically as a anti-rheumatic disease, anti-stress, ischemic heart disease and gastric ulcer. In the present study, we examined the adjuvant activity of the crude polysaccharides fraction from Eleutherococcus senticosus, EN-3, on the induction of humoral and cellular immune responses against bovine serum albumin (BSA) or ovalbumin (OVA). The thioglycollate-induced macrophages and silica-induced dendritic-like cells cultured with BSA and EN-3 synergistically increased the production of TNF-$\alpha$ and IL-12. When mice were subcutaneously immunized with BSA + EN-3, the antibody titer against BSA was showed significantly higher than those immunized with BSA alone. In addition, when mice were immunized with OVA + FIA + EN-3, the antibody titer was showed similar patterns with the FCA. The assay for determining subisotype of antibody revealed that EN-3 augmented OVA-specific antibody titer of IgG1 and IgG2b. The culture supernatant obtained from splenocytes of mice treated with OVA + FIA + EN-3 also showed a higher level of both OVA-specific Th1-type (IL-2, IFN-${\gamma}$ and GM-CSF) and Th2-type cytokine (IL-4, IL-6 and IL-10). In vitro analysis of T cell proliferation to OVA on 8 weeks, the splenocytes of mice treated with OVA + EN-3 showed a significantly higher proliferating activity than those treated with OVA alone. These results suggest that EN-3 may possess adjuvant activities to potentially to enhance humoral as well as cellular immune response.

• Macromolecular Research
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• 제17권2호
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• pp.128-132
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• 2009

Fucoidan, a sulfated polysaccharide derived from brown seaweed, is an important material valued for its various biological functions, including anti-coagulation, anti-aging, and immune system support. In this study, we examined the potential of fucoidan as a novel emulsifying agent in BSA (bovine serum albumin)-stabilized emulsion at a neutral pH. We measured the dispersed oil-droplet size, surface zeta-potential and creaming formation of 0.5 wt% BSA emulsion (20 wt% oil traction) in the absence and presence of fucoidan. The average particle size and zeta-potential value were 625.4 nm and -30.91 mV in only BSA-stabilized emulsion and 745.2 nm and -44.2 mV in 1.0 wt% fucoidan-added BSA emulsion, respectively. This result suggested that some positive charges of the BSA molecules interacted with the negative charges of fucoidan to inhibit the flocculation among the oil droplets. The creaming rate calculated from the backscattering data measured by Turbiscan dramatically decreased in 1.0 wt% fucoidan-added BSA emulsion during storage. Accordingly, the repulsion forces induced among the oil particles coated with 1.0 wt% fucoidan in emulsion solution resulted in significantly increased emulsion stability. The turbidity of the BSA-stabilized emulsion at 500 nm decreased during five days of storage. However, the fucoidan-added BSA emulsion exhibited a higher value of turbidity than the BSA-stabilized emulsion did. In conclusion, an anionic sulfated fucoidan lowered the surface zeta-potential of BSA-coated oil droplets via the electrostatic interaction, and subsequently inhibited the flocculation among the oil droplets, thereby clearly minimizing the creaming and phase separation of the emulsion.

• 한국막학회:학술대회논문집
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• 한국막학회 1998년도 춘계 총회 및 학술발표회
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• pp.99-101
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• 1998

1. Introduction : The conventional grafting polymerization technique requires chemically reactive groups on the surface as well as on the polymer chains. For this reason, a series of prefunctionalization steps are necessary for covalent grafting. The surface prefunctionalizational technique for grafting can be used to ionization radiation, UV, plasma, ion beam or chemical initiators. Of these techniques, radiation method is one of the useful methods because of uniform and rapid creation of active radical sites without catalytic contamination in grafted samples. If the diffusion of monomer into polymer is large enough to come to the inside of polymer substrate, a homogeneous and uniform grafting reaction can be carried out throughout the whole polymer substrate. Radiation-induced grafting method may attach specific functional moieties to a polymeric substrate, such as preirradiation and simultaneous irradiation. The former is irradiated at backbone polymer in vacuum or nitrogen gas and air, and then subsequent monomer grafting by trapped or peroxy radicals, while the latter is irradiated at backbone polymer in the presence of the monomer. Therefore, radiation-induced polymerization can be used to modification of the chemical and physical properties of the polymeric materials and has attracted considerable interest because it imparts desirable properties such as blood compatibility. membrane quality, ion excahnge, dyeability, protein adsorption, and immobilization of bioactive materials. Synthesizing biocompatible materials by radiation method such as preirradiation or simultaneous irradiation has often used $\gamma$-rays to graft hydrophilic monomers onto hydrophobic polymer substrates. In this work, in attempt to produce surfaces that show low levels of anti-fouling of bovine serum albumin(BSA) solutions, hydroxyethyl methacrylate(HEMA) was grafted polypropylene membrane surfaces by preirradiation technique. The anti-fouling effect of the polypropylene membrane after grafting was examined by permeation BSA solution.

• Journal of Pharmaceutical Investigation
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• 제38권6호
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• pp.357-363
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• 2008

Penetration rate of large molecule through skin is very low due to the barrier effect of stratum corneum. Novel microneedle treatment device with roll was designed for transdermal delivery of large molecular drugs such as vaccine and protein drugs. The permeation rates of FITC labelled bovine serum albumin (FITC-BSA) as a model protein were determined using modified Franz diffusion cell and hairless mouse skin which were treated by hydrogel or solution containing FITC-BSA. Fluorescent spectrophotometer was used to analyze the concentration of FITC-BSA. Microscope using fluorescent filter was used to capture the image and location of FITC-BSA in the skin. We confirmed that permeation rate of BSA was increased with the treatment by microneedle and was increased by the increasing frequency of treatment. Furthermore, the permeation rate observed from hydrogel treated skin was significantly higher than that from solution treated skin.

• Journal of Microbiology and Biotechnology
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• 제6권1호
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• pp.1-6
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• 1996

A bacterial strain NS70 producing an alkaline protease was isolated from soil samples taken near a hot spring and identified as Bacillus licheniformis by its morphological and physiological properties and cellular fatty acid analysis. The isolated alkaline protease was purified by ammonium sulfate fractionation, DEAE-, CM-, and Phenyl-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 32, 000 Da by sodium dodecylsulfate polyacrylamide gel electrophoresis. Its optimal pH and temperature for proteolytic activity against Hammarsten casein were 12 and $65^{\circ}C$, respectively. The enzyme was stable at alkaline pH range from 6.0 to 12.0, and fairly stable up to $65^{\circ}C$. The enzyme was inhibited by phenylmethylsulfonyl fluoride but not by EDTA and N-ethylmaleimide indicating that the enzyme is serine protease. Enzyme activity was markedly inhibited by $Hg^{2+}$ and $Cu^{2+}$. Autolytic phenomena were observed on purified protease NS70 but autolysis was reduced by the addtion of $Ca^{2+}$ ion or bovine serum albumin.

• 한국식품과학회지
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• 제22권1호
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• pp.56-60
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• 1990

역류분배 방법을 이용한 단백질의 분리정제 가능성을 높이기 위하여 4가지 모델단백질에 대하여 수용성 액체 2상계에서의 분획계수를 pH 변화에 따라 조사하였다. 또한 역류분배를 이용한 다중분배추출방식으로 모델 단백질의 혼합물을 수준의 pH(4.5-12.0)에서 분획하였다. 그 결과 pH에 따라 단백질들의 등전점 차이로 인한 분획계수(K)값이 다르게 나타서, 다중분배를 행할 때 각 단백질 분획들이 위치하는 튜브번호가 다르게 되므로 역류분배 방법을 통한 단백질의 분리 가능성을 보여주었다.

• Archives of Pharmacal Research
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• 제10권4호
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• pp.223-227
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• 1987

The effects of butylated hydroxyanisole and butylated hydroxytoluene on the immune status in normal male were evaluated. They exhibited significant decrease in the circulating leukocyte counts. Relative spleen and thymus weights were slightly decreased, but not stratistically significant. These were, however, significant liver hypertrophies in theier exposed mice. Splenic IgM PFCs per one million cells in 1/20 LD50 BHA and BHT exposed mice were significantly reduced IgM PFCs per spleen were similar tothose of control, except in 1/20 LD50 BHA exposed mice, where they were significantly suppressed. The precise nature of the inhibition is not clear. Direct cytotoxicity is not responsible for the depressed antibody response, even following relatively high doses of them, because the changes in spleen cellularity are not significant. Both substances, however, did not show any effects on the arthus reaction and delayed hypersensitivity reaction induced by heat aggreagted bovine serum albumin, and in vivo phagocytosis of colloidal carbon. In the light of the present results, in vivo antibody response as well as in vitro, may be sensitive to BHA of the present results, in vivo antibody response as well as in vitro, amy be sensitie to BHA and BHT. Further elucidation of the precise nature of antibody suppression in their exposed mice, is warranted.