• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.615-618
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• 2000

The purpose of present study was to examine the efficacy of PAB (para-amino benzamidine) affinity column chromatography, pasteurization ($60^{\circ}C$ heat treatment for 10 h), and lyophilization steps, employed in the manufacture of urokinase from human urine, in the removal and/or inactivation of urine-born viruses. Bovine herpes virus (BHV) and Murine encephalomyocarditis virus (EMCV) were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses and the amount of virus in each fraction was quantified by 50% tissue culture infectious dose ($TCID_{50}$). BHV and EMCV were effectively partitioned from urokinase during PAB chromatography with the log reduction factors of 6.71 and 5.27, respectively. Pasteurization was a robust and effective step in inactivating BHV and EMCV, of which titers were reduced from initial titers of $8.65\;log_{10}\;TCID_{50}$ and $7.81\;log_{10}\;TCID_{50}$, respectively, to undetectable levels within 1 hour of treatment. The log reduction factors achieved during lyophilization were 2.06 for BHV and 4.54 for EMCV. These results indicate that the production process for urokinase has sufficient virus reducing capacity to achieve a high margin of virus safety.

• Journal of the Korean Magnetic Resonance Society
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• v.15 no.2
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• pp.104-114
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• 2011

Rapid exchange and hydrolysis of the tubulin-bound guanine nucleotides have been known to govern the dynamics of microtubules. However, the instability and low concentration have made it difficult for the microtubule-bound GTP to be observed directly. In this study, we circumvent these problems by lyophilization and using cross-polarization techniques. $^{31}P$ NMR signals were detected from the tubulin-bound GTP in microtubules for the first time. Analysis of the $^{31}P$ CPMAS NMR spectrum indicates that GTP hydrolysis was delayed by the presence of taxol.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.18 no.3
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• pp.371-377
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• 1996

Bone graft has been used to repair one defect caused by disease and trauma, congenital and acquired deformities. Graft materials are autogenous bone, allogenic bone, xenogenic bone, synthetics. Autogenous bone graft is the most superior to other materials for immunologic reaction, compatibility to host tissue, and revascularization. However, autogenous bone graft is required for additional operation and the amount of taking is limited. Autografts are obtained at own expense and also limited in size, shape. In order to compensate these problems, allogenic bone graft has been used increasingly. But allogenic bone graft encounters immunologic complications. Therefore, it has been used after freezing, lyophilization, or demineralization. Allogenic bone processed by only lyophilization includes potential antigenic properties on its surface, therefore it is demineralized to deplete immunologic reaction. Demineralized bone releases BMP and helps the mesenchymal cells transform to the chondroblast to produce cartilage and bone. This reaction is called osteoinducation. Many authors have reported that mineralized lyophilized bone had less antigenicity clinically and favorable bony consideration with host bone. In our department from 1995 to now, we have used banked allogenic bone graft that has been prepared from Wonkwang Bone Bank in 5 cases and mineralized lyophilized bone graft in 2 cases to reconstruct the maxillofacial bone defect after tumor resection and cyst enucleation and cleft alveolus. We will report with literature review that the result is favorable functionally and esthetically.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.19-25
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• 2006

With particular regards to the hepatitis A virus (HAV), a terminal dry-heat treatment ($100^{\circ}C$ for 30 min) process, following lyophilization, was developed to improve the virus safety of a solvent/detergent-treated antihemophilic factor IX concentrate. The loss of factor IX activity during dry-heat treatment was of about 3%, as estimated by a clotting assay. No substantial changes were observed in the physical and biochemical characteristics of the dry-heat-treated factor IX compared with those of the factor IX before dry-heat treatment. The dry-heat-treated factor IX was stable for up to 24 months at $4^{\circ}C$, The dry-heat treatment after lyophilization was an effective process for inactivating viruses. The HAV and murine encephalomyocarditis virus (EMCV) were completely inactivated to below detectable levels within 10 min of the dry-heat treatment. Porcine parvovirus (PPV) and bovine herpes virus (BHV) were potentially sensitive to the treatment. The log reduction factors achieved during lyophilization and dry-heat treatment were ${\ge}5.60$ for HAV, ${\ge}6.08$ for EMCV, 2.64 for PPV, and 3.59 for BHV. These results indicate that dry-heat treatment improves the virus safety of factor IX concentrates, without destroying the activity. Moreover, the treatment represents an effective measure for the inactivation of non-lipid enveloped viruses, in particular HAV, which is resistant to solvent/detergent treatment.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.997-1003
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• 2008

Viral safety is a prerequisite for manufacturing clinical antihemophilic factor VIII concentrates from human plasma. With particular regard to the hepatitis A virus (HAV), a terminal dry-heat treatment ($100^{\circ}C$ for 30 min) process, following lyophilization, was developed to improve the virus safety of a solvent/detergent-treated antihemophilic factor VIII concentrate. The loss of factor VIII activity during dry-heat treatment was of about 5%. No substantial changes were observed in the physical and biochemical characteristics of the dry-heat-treated factor VIII compared with those of the factor VIII before dry-heat treatment. The dry-heat-treated factor VIII was stable for up to 24 months at $4^{\circ}C$. The dry-heat treatment after lyophilization was an effective process for inactivating viruses. The HAV, murine encephalomyocarditis virus (EMCV), and human immunodeficiency virus (HIV) were completely inactivated to below detectable levels within 10 min of the dry-heat treatment. Bovine herpes virus (BHV) and bovine viral diarrhea virus (BVDV) were potentially sensitive to the treatment. However porcine parvovirus (PPV) was slightly resistant to the treatment. The log reduction factors achieved during lyophilization and dry-heat treatment were ${\geq}5.55$ for HAV, ${\geq}5.87$ for EMCV, ${\geq}5.15$ for HIV, 6.13 for BHV, 4.46 for BVDV, and 1.90 for PPV. These results indicate that dry-heat treatment improves the virus safety of factor VIII concentrates, without destroying the activity. Moreover, the treatment represents an effective measure for the inactivation of non-lipid-enveloped viruses, in particular HAV, which is resistant to solvent/detergent treatment.

• Proceedings of the PSK Conference
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• 2000.10a
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• pp.260.2-261
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• 2000

• Journal of Pharmaceutical Investigation
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• v.31 no.3
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• pp.167-172
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• 2001

All-trans retinoic acid (ATRA), vitamin A acid, has been shown to exert anticancer activity in a number of types of cancers, particularly in acute promyelocytic leukaemia (APL). Due to its highly variable bioavailability and induction of its own metabolism after oral treatment, development of parenteral dosage forms are required. However, its poor aqueous solubility and chemical unstability give major drawbacks in parenteral administration. This study was undertaken to investigate a possibility to develop a parenteral formulation of ATRA by employing solid lipid nanoparticle (SLN) as a carrier. By optimizing the production parameters and the composition of SLNs, SLNs with desired mean particle size (<100 nm) as a parenteral dosage form could be produced from trimyristin (as solid lipid), Egg phosphatidylcholine and Tween 80 (as SLN stabilizer). The mean particle size of SLN formulation of ATRA was not changed during storage, suggesting its physical stability. Thermal analysis confirmed that the inner lipid core of SLNs exist at solid state. The mean particle size of ATRA-loaded SLNs was not significantly changed by the lyophilization process. ATRA could be efficiently loaded in SLNs, while maintaining its anticancer activity against HL-60, a well-known APL cell line. Furthermore, by lyophilization, ATRA loaded in SLN could be retained chemically stable during storage. Taken together, our present study demonstrates that physically and chemically stable ATRA formulation adequate for parenteral administration could be obtained by employing SLN technology.

• Macromolecular Research
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• v.14 no.1
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• pp.94-100
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• 2006

Although the peripheral nerve system has a relatively good regenerating capacity compared to the central nerve system, peripheral nerve repair remains a clinical challenge as restoration of normal nerve function is highly variable. Synthetic tubular nerve conduits were designed as an alternative repair method in order to replace the need for an isograft. These nerve conduits guide regenerating axons from the proximal toward the distal end, maintain within growth-promoting molecules released by the nerve stumps, and protect regenerating axons from infiltrating scar tissue. In this work, we prepared cinnamoylated alginate (CA)-collagen-chitosan nerve conduit using the lyophilization method to generate a controllable parallel channel in the center and then investigated its influence on peripheral nerve regeneration in an animal study. At 12 weeks after implantation, histological study showed that tissue cable was continuously bridging the gap of the sciatic nerve in all rats. Our newly developed nerve conduit is a promising tool for use in peripheral nerve regeneration and provides a suitable experimental model for future clinical application.

• Archives of Pharmacal Research
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• v.26 no.6
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• pp.504-510
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• 2003

The aim of this study is to prepare biodegradable microspheres without the use of surfactants or emulsifiers for a novel sustained delivery carriers of protein drugs. A poly($\varepsilon$-caprolactone)/poly(ethylene glycol)/poly($\varepsilon$-caprolactone) (CEC) triblock copolymer was synthesized by the ring-opening of $\varepsilon$-caprolactone with dihydroxy poly (ethylene glycol) to prepare surfactant-free microspheres. When dichloromethane (DCM) or ethyl formate (EF) was used as a solvent, the formation of microspheres did not occur. Although the microspheres could be formed prior to lyophilization under certain conditions, the morphology of microspheres was not maintained during the filtration and lyophilization process. Surfactant-free microspheres were only formed when ethyl acetate (EA) was used as the organic solvent and showed good spherical micro-spheres although the surfaces appeared irregular. The content of the protein in the micro-sphere was lower than expected, probably because of the presence of water channels and pores. The protein release kinetics showed a burst release until 2 days and after that sustained release pattern was showed. Therefore, these observations indicated that the formation of microsphere without the use of surfactant is feasible, and, this the improved process, the protein is readily incorporated in the microsphere.

• YAKHAK HOEJI
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• v.43 no.6
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• pp.793-797
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• 1999

Salmonella typhi Ty21a is an attenuated strain of S. typhimurium and used for oral typhoid vaccine. In an attempt to increase the stability of Ty21a manufacturing typhoid vaccine, we studied about the stability of freeze-dried Th21a including additives at various temperature conditions. In order to investigate the freeze-drying rate of Ty21a according to various absorbance, we lyophilized Ty21a by using 8% sucrose as a stabilizer. The optimal freeze-drying rate of Ty21a was appeared when OD (optical density) value of the growth was between 2.5 and 3.0. To investigate the stability of Ty21a at various temperature, the viability was measured after storaging the freeze-dried Ty21a at the room temperature, cold and freezing condition for 1 week. The viability of Ty21a in cold and freezing storage condition was 5 times more stable than in room temperature. To search the most stable additives for the freeze-dried Ty21a, the viability of Ty21a including additives at the various storage condition was estimated. Mannitol and loctose were the most stable additives. Theses results suggest that the OD value of Ty21a growth, low temperature, mannitol and lactose are important factors for the optimal freeze-drying rate, the stable storage and the most stable additives, respectively.