• Journal of Microbiology and Biotechnology
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• 제27권4호
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• pp.718-724
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• 2017

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

• Journal of Pharmaceutical Investigation
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• 제34권3호
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• pp.177-183
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• 2004

Recently, studies on intranasal mucosa delivery of influenza vaccine have been actively developed because of lack of pain and ease of administration. We studied on preparation of nanoparticle delivery system using biodegradable polymer as a poly(DL-lactide-co-glycolide) (PLGA) and their binding characteristics with vaccine. Three kinds of PLGA nanoparticles were prepared by spontaneous emulsification solvent diffusion (SESD) method using sodium dodecyl sulfate and sodium laurate as an anionic surfactant and Lutrol F68 (polyethylene glycol-block-polypropylene glycol copolymer) as a nonionic surfactant. The 5-aminofluorescein labeled vaccine was coated on the surface of nanoparticles by ionic complex. The complexes between vaccine and nanoparticles were confirmed by change of the size. After vaccine coating on the surface of anionic nanoparticles, particle size was increased from 174 to 1,040 nm. However the size of nonionic nanoparticles was not more increased than size of anionic nanoparticles. The amount of coated vaccine on the surface of PLGA nanoparticles was $14.32\;{\mu}g/mg$ with sodium dodecyl sulfate, $12.41\;{\mu}g/mg$ with sodium laurate, and $9.47{\mu}g/mg$ with Lutrol F68, respectively. In conclusion, prepared nanoparticles in this study is possible to use as a virus-like nanoparticles and it could be accept in the field of influenza vaccine delivery system.

• 대한수의학회지
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• 제12권1호
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• pp.71-75
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• 1972

Due to the fact that an inactivated anthrax vaccine may show no or lower immunogenicity and stability, a number of spore vaccines were exclusively used worldwide. In these studies non or less allergic strain of anthrax bacillus was selected and made a capsulated vegetative organisms. Anthrax organisms of a virulent strain were cultivated on sodium bicarbonate medium with or without adding I-alanine in which B. anthracis grew luxuriantly without forming spores. Inactivation of the organisims was carried out at $37^{\circ}C$ water bath for 3 days after the bacterial culture was mixed with formalin in a final concentration of two per cent. Aluminum hydroxide gel was added to the mixture of anthrax and blackleg bacterin. Guinea pigs were injected with the vaccine via subcutaneous or intramuscular route and challenged after three weeks, and the possibilities of protection was tested. Throughout the studies, the vaccines possibly protected the vaccinated guinea pigs more than 80 per cent compared to that of the controls. This experimental results strongly suggest that the vaccine may possibly applicable to the prevention of bovine anthrax and blackleg.

• Journal of Pharmaceutical Investigation
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• 제31권4호
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• pp.241-256
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• 2001

Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.

• KSBB Journal
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• 제19권1호
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• pp.88-92
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• 2004

장티푸스 협막 다당체와 일본 뇌염 바이러스로 구성되어있는 혼합백신을 제조하여 마우스에서 면역성을 측정하였다. 혼합 항원 백신은 단일 항원을 투여한 경우보다 장티푸스와 일본 뇌염에 대해서 더 높은 IgG 항체 형성을 나타내었다. Aluminium hydroxide를 adjuvant로 첨가한 경우에도 첨가하지 않은 대조군에 비하며 모두 IgG 항체 형성이 증가하는 것으로 나타났다. 일본뇌염의 경우 중화항체 값 측정 시험에서도 혼합 항원백신은 일본뇌염 단독 항원 시에 비교해서 더 많은 중화항체 값을 형성하는 결과를 나타내었다. 본 실험에서 장티푸스 협막 다당체와 일본뇌염 바이러스의 혼합백신은 서로 다른 성분의 항원간에 나타날 수 있는 항체 형성에 대한 masking effect가 발생하지 않으며 오히려 synergic effect를 나타낸다는 사실을 보여주었다. 본 연구에서는 접종시기가 유사한 두 항원의 혼합 백신으로의 사용가능성을 높여주고 최근에 백신 산업에서 종류가 다른 여러 백신들을 혼합하여 다가백신을 제조 생산하는 추세의 산업화에 큰 기대효과를 제공할 수 있으리라 사려된다.

• Toxicological Research
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• 제32권4호
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• pp.269-274
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• 2016

The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.

• 수산해양교육연구
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• 제26권6호
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• pp.1366-1372
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• 2014

본 연구의 목적은 해산어류와 갑각류에 심각한 위해를 끼치고 있는 V. harveyi에 대한 백신개발을 위하여 V. harveyi 백신생산에 적합한 배양배지를 탐색하고 적정 투여량을 조사하며 또한 S. parauberis 백신과 혼합백신의 형태로 투여시 백신효능에 대해 조사하는 것이다. 이를 위하여 2-2'-dipyridyl이 첨가되거나 첨가되지 않은 TSB와 BHIB에 배양 후 FKC 백신을 제작한 후 넙치에 투여하여 응집항체가의 생산변화와 공격실험에서의 상대생존율을 비교분석하였다. 또한 V. harveyi 백신의 적정 투여량을 정하기 위하여 어체중 kg 당 10mg 또는 20mg을 투여하여 면역반응을 비교하였으며 S. parauberis 백신을 혼합한 백신을 투여한 후 면역반응을 비교하였다. 그 결과 2-2'-dipyridyl이 들어간 TSB와 BHIB에 배양된 V. harveyi 백신은 응집항체형성과 방어력에서 차이를 보이지 않았다. 또한 백신 투여량에 따른 응집항체가에 있어서 큰 차이는 없었으나 어체중 kg 당 10mg을 투여한 실험구가 조금 높은 방어력을 나타내었다. S. parauberis 백신과 혼합한 dual 백신을 투여시 V. harveyi 백신만을 단독으로 투여했을 때와 비교해 방어력이 현저히 증가하였으며 특히 어체중 kg당 두가지 백신을 각각 10mg씩 혼합하여 투여한 실험구에서는 28일동안 폐사가 전혀 일어나지 않아 백신효능이 매우 뛰어난 것으로 나타났다. 결론적으로 넙치를 위한 V. harveyi 백신을 개발할 때는 2-2'-dipyridyl가 첨가된 TSB에 배양 후 제작된 V. harveyi 백신을 어체중 kg당 10mg의 투여량으로서 S. parauberis 백신과 혼합투여하는 것이 효능과 경제적인 면에서 바람직하다고 생각된다.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.214.2-215
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• 2000

• 보건행정학회지
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• 제25권2호
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• pp.140-148
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• 2015

Vaccination is the most powerful and useful preparation against infectious diseases. However, developing vaccines costs a lot and requires extensive long-term efforts. Therefore, the government should research and develop vaccines with a national-level policy. To greatly enhance the success rate of vaccine development, the policy should be set up considering priorities such as the current status of domestic research, the importance for public health, the urgency of research. The Delphi technique was utilized to draft this survey, through a brainstorming stage, then two inquiries, and finally the final panel meeting where unresolved items were discussed, to draw the conclusion. Among the results, firstly, the highest ranked item on centralized fields for vaccine development by the Ministry of Health was 'self-sufficiency of vaccines.' Secondly, 'emerging infectious disease' was most highly ranked in prioritized fields of vaccine development and research. Thirdly, for the vaccine that needs to be improved and developed further by the government to improve its efficacy and safety, BCG (Bacille de Calmette) for tuberculosis was ranked the highest on both types (intradermal and subcutaneous injection) from National Immunization Programme (NIP) and non-NIP. As for the high risk pathogens, 'anthrax' and 'smallpox' were first and second, consecutively. Lastly, 'development and control of vaccine candidates' was ranked the highest for the area in need for technique development in order to improve domestic vaccine's research level. The results of this study will be put to good use as basic data for the national vaccine research and development (R&D) policy of the country. This study was first step and more studies should be carried out for the final decision of the national vaccine R&D priority.

• 대한의생명과학회지
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• 제12권3호
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• pp.177-183
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• 2006

To demonstrate the effect of formulation conditions and evaluations of structural integrity from ovalbumin containing poly lactide glycolide copolymer (PLGA) microspheres for Vaccine delivery, OVA microspheres were prepared by a W/O/W multiple emulsion solvent extraction technique. Dichloromethan (DCM) and Ethyl acetate (EA) were applied as an organic phase and poly vinyl alcohol (PVA) as a secondary emulsion stabilizer. Microspheres were characterized for particle size, morphology (optical microscopy and Scanning Electron Microscope (SEM)). Protein denaturation was evaluated by size exclusion chromatography (SEC), SDS-PAGE and isoelectric focusing (IEF). Residual organic solvent was estimated by gas chromatography (GC) and differential scanning calorimetry (DSC). Optical photomicrograph and SEM revealed that micro spheres were typically spherical but various morphologies were observed. Mean particle size $(d_{vs})$ of microspheres were in the range of $3{\sim}50{\mu}m$. Also, The protein stability was not affected by the fonnulation process and residual organic solvent was beyond the detection below 0.1ppm. These results demonstrated that micro spheres might be a good candidate for the parenteral vaccine delivery system.