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The specificity of enzymes promises great improvements in a variety of their potential applications. However, the short lifetimes of enzymes frequently limit their usefulness. Recent breakthroughs in nanotechnology have made various nanostructured materials more affordable for a broader range of applications, including enzyme immobilization and stabilization [1-3]. This presentation will discuss recent developments in nanobiocatalysis to improve the enzyme stability using various nanostructures such as single enzyme nanoparticles (SENs), mesoporous materials, nanofibers, nanoparticles, and carbon nanotubes. In the form of SENs, each enzyme molecule is surrounded with a nanometer scale network, resulting in stabilization of enzyme activity without any serious limitation for the substrate transfer from solution to the active site [4]. The approach of nanometer-scale enzyme reactors (NERs) stabilized enzymes in mesoporous media via a ship-in-a-bottle effect, which employs adsorption of enzymes followed by enzyme crosslinking [5]. A similar approach resulted in the enzyme coating on the surface of electrospun nanofibers, which also stabilized the enzyme activity in a vivid way [6-7]. Stabilized enzyme systems in various nanostructures will make an ideal system for many applications including bioremediation, antifouling, biosensors, bioreactors, microfluidic devices, trypsin digestion, and biofuel cells. Several successful examples of nanobiocatalytic applications will be given in this presentation.

The living organisms detect various stimuli from the environment, process these environmental information to recognize their circumferences, and make behavioral decision. The chemosensory information including gustatory and olfactory information is most important for the living organisms to survive the harsh environment. The models describing the highly efficient environmental responding mechanisms of living organisms can be used as control algorithms in the field of systems engineering. To realize this biomimetic intelligent control, we require a stripped-down model that expresses a series of information processing tasks from stimulation input to movement. We selected Paramecium caudatun as a target organism because it has a relatively simple organizational structure, which can be characterized using biochemical, behavioral and electrophysiological analyses. We particularly focused on a motility response known as chemotaxis and developed a computer model that includes not only lectrophysiological information processing but also cilia control. After confirming the effectiveness and validity of the proposed model by a series of computer simulations, we applied it to a mobile robot control problem. Furthermore, we analyzed the odor discrimination of the mice by Y-maze behavioral assay to obtain a new knowledge on the olfactory information process system. The mice were able to discriminate the odor of learned red wine from the odors of other liquor except for the different brand red wine with the similar odorant component like the sommelier. In the case to discriminate learned red wine from the different brand red wine, the responses were divided into 3 groups, discriminate correctly, discriminate incorrectly, cannot discriminate. These individualities were also observed in the behavior analysis using known pure substance. These results show that the mice discriminate the odors by attending to the part of odor information which received the mouse, and that the selection of the information called “selective attention” which recognized in visual sense and auditory system take place in the olfactory system. Olfactory stimulus is composed of a combination of 400 thousands kinds of odorant molecules. Processing all olfactory information is usually a time-consuming way. When the source of the odor is associated with natural enemies or harmful substances, this delay may become fatal to living organisms. Selective attention would be able to reduce the dimension of olfactory information and information processing time. Furthermore, selective attention in olfactory system could be modified through the learning experiences. Elucidating the algorithm of olfactory selective attention would be effective for an application to the control algorithm for the systems with a large number of observable variables.

Chromosome- and genome-engineering technologies are becoming increasingly important in biotechnology as well as genome science. Since they have provided powerful approaches to analyze functions of large chromosomal regions or many genes simultaneously and are expected to contribute to breeding of novel microorganisms for a new phase of efficient production of valuable biomaterials, development of a simple and efficient technology for manipulating chromosome and genome is greatly demanded for promoting both basic and applied biosciences. As one of the key technology for manipulating chromosome and genome, we have developed a novel chromosome-engineering technology which we called chromosome-splitting technology. Chromosome-splitting technology is a procedure to cut a chromosome, at any chosen site, into two pieces, but to make them behave as functional chromosomes. Chromosomes must have three elements for their maintenance during mitotic growth and meiotic development, i.e., centromere, telomere, and replication origin. Therefore, if chromosomes are newly generated, whether derived from natural or artificially constructed, they must have these three elements for faithful transmission to daughter cells. Because the Saccharomyces cerevisiae has served as a valuable model eukaryote and also as an extremely useful industrial microorganism, we initiated work in this organism. We have developed the chromosome-splitting method that enables very simple, efficient, and repeatable splitting of chromosome1). This new method was designated as PCS (PCR-mediated Chromosome Splitting) method. The PCS method combines a streamlined procedure (two-step PCR and one transformation per splitting event) with the Cre/loxP site-specific recombination system for marker rescue. The PCS method can be used for a wide variety of purposes such as: (1) chromosome shuffling to swap a particular chromosomal region from one strain with the corresponding region from another for exploring genotypephenotype relationships at the sub-chromosomal level, (2) genome reconstruction to create novel strains with a variety of genomic constitutions, (3) one-step deletion of a designed internal or terminal chromosomal region to survey functions of genes in defined chromosomal regions, and (4) application for the manipulation and functional analysis of chromosome fragments of higher eukaryotes cloned into yeast artificial chromosomes2-12). These advances to manipulate chromosome and genome in a large scale are expected to accelerate the breeding of novel strains for biotechnological purposes, and also to reveal functions of presently uncharacterized genes and chromosomal regions in S. cerevisiae.

“Shochu kasu”, a by-product of shochu production, is discharged during the distillation process. In recent years, the approximate amount of shochu kasu formed in the Japanese shochu industry has exceeded 850,000 tons per year. A large amount of shochu kasu is still treated as an industrial waste. Therefore, an effective process for utilizing shochu kasu is desired. Our aim in this study was to develop novel conversion processes employing lactic acid bacteria from shochu kasu into valuable materials in order to promote its recycling. Fermented barley extract (FBE) was obtained from shochu kasu by physical isolation (vibrating sieve, screw press, ceramic filtration). FBE was ineffective as a carbon source because most of the digestible carbon sources derived from barley were already utilized by yeast and fungi (koji) during shochu production. However, when FBE was evaluated as a nitrogen source by comparing it with a nutritionally rich laboratory-use nitrogen source (yeast extract etc.) in the basal medium, the FBE medium yielded higher rates and maxima of cell growth of Lactobacillus fermentum NBRC 3071 and Bifidobacterium longum JCM 1217T than the basal medium. Despite the inhibitory effect in the initial phase, the final cell growth level of Lactococcus lactis subsp.lactis ATCC 11454 and Enterococcus faecalis NCIMB 8275 in the FBE medium reached to the same level as that in the basal medium. Ethanol insoluble (EI) fraction of FBE had a growth-stimulating effect on lactic acid bacteri a and bifidobacteria. EI fraction was subjected to reversed phase and gel-filtration chromatography to get two active fractions, of which the main component was oligosaccharide consisted of glucose, xylose, and arabinose and the minor one was peptide. As valuable substances produced by lactic acid bacteria, we focused on “nisin A” (the only bacteriocin widely used as a food preservative) and “　-amino butyric acid (GABA)” (a nonprotein amino acid having several physiological functions such as hypotensive, diuretic, tranquilizing and so on). L. lactis subsp. lactis ATCC 11454 and Enterococcus sp. FC 301 isolated from the French dairy products were used as nisin A producer and GABA producer, respectively. The FBE medium supplemented with glucose provided a high level of nisin A and GABA production with yields comparable to those from a nutritionally rich laboratory medium (the basal medium). By adding the EI fraction to the basal medium, nisin A production was markedly enhanced. A large scale production test of nisin A and GABA performed in the pilot plant (3-t fermentor) gave the same productivity as that in lab scale. Based on the procedure developed in this study, we have already succeeded in commercializing “Fermented barley GABA” (containing high concentration of GABA) and “Barley-X” (inexpensive practical culture medium)

Genomics-guided technologies including cDNA microarrays have been successfully applied for the identification of gene expression alterations in various Streptomyces strains. To detect global changes in mRNA abundance associated with overproduction of anti-cancer doxorubicin, reverse engineering strategy via comparative transcriptome analyses in cultures of the wild-type and industrial mutant strains of S. peucetius were conducted using interspecies S. coelicolor cDNA microarrays. Among 160 S. coelicolor potential candidate genes showing at least a two-fold change in transcription profile, a SCO3579 (whiB-like gene named wblA) inhibited the biosynthesis of doxorubicin in S. peucetius as well as the production of antibiotics in S. coelicolor, suggesting that wblA and its homologs act globally among streptomycetes as down-regulators of antibiotic biosynthesis. In addition, one of tetR-family transcriptional regulatory genes was also identified to encode a global antibiotic regulator. Gene disruption of S. coelicolor revealed that this tetR-family regulatory gene down-regulates antibiotic biosynthesis through pathway-specific regulators even in the presence of the wblA transcript, implying that it might encode a wblA-bypassing antibiotic down-regulator. To isolate and manipulate a wblA homolog in a doxorubicin-overproducing S. peucetius industrial stain, a total genomic DNA library from a S. peucetius industrial mutant was constructed and screened using a S. coelicolor wblA as a probe, resulting in isolation of a wblA ortholog (wblA-spe) showing 95% amino acid identity to a S. coelicolor wblA. Gene disruption of wblA-spe from the S. peucetius industrial mutant resulted in additional increase of doxorubicin production, implying that doxorubicin productivity in the S. peucetius industrial mutant strain could be further improved via rational comparative transcriptomics-guided target gene manipulation.

High cell density in microbial cell cultivation is about 10 times to a normal density that can be obtained without extraordinary efforts in a batch culture. For instance, if a 5~10g/L of E.coli cell density is considered a normal cell density, 50~100g/L would be called a high cell density. In animal cell culture 106~107/ml cell density would be a high cell density if 105~106 cells/ml. An immobilized cell matrix can have a cell density of 108cells/ml, but the cell density in the reactor can be lower or much lower than that in the matrix. Extra efforts mean fed-batch, continuous culture, membrane cell recycle for high cell density culture and use of pure oxygen. Several HCDC systems developed at biochemical engineering laboratory of KAIST during the last 30 years will be introduced with a theoretical simulation showing the equivalence of MSC-HCDC with similar titer to fed-batch but with much higher productivities in lactic acid and monoclonal antibody production. Especially depth filter perfusion system (DFPS) for monoclonal antibody production appears to be suitable for MSCHCDC pilot test because of its simplicity in immobilization and no need in separation of cells from the product stream. MSC-HCDC: multi-stage continuous high cell density culture

21세기가 생명공학기술의 세기가 될 것이라고 예언했던 1999년도 타임지의 보도이후 만 10년이 지난 2009년 말 오늘의 세계경제는 예상보다도 더 빠르게 생명공학 기술의 발전과 산업의 성장이 이루어지고 있다. 주지하는 바와 같이 세계는 인구의 고령화, 웰빙 풍조의 확산, 자원 고갈 및 기후변화 등의 글로벌 추세에 맞추어 어느 때보다도 바이오산업의 성장 가능성을 높이 평가하고 있다. 그러나 최근의 세계적 금융위기에서 목도하는 바와 같이 전혀 예상하지 못했던 외적 경제변수의 등장으로 신산업으로서 바이오산업의 위상도 투자가 격감하는 등 더불어 위축되는 현상도 감지되고 있다. 이러한 세계적 추세 속에서 실제로 어떠한 생명공학 기술, 어떠한 바이오 사업이 새로운 투자를 끌어들이는 역할을 할 수 있을지에 대해 많은 관심이 기울여지고 있다. 본 강의에서는 글로벌 추세와 바이오기술의 진보 및 발전 동향에 대해 다시 한번 짚어보고 이를 기반으로 한 바이오산업의 기회와 도전에 대해 논의해보고자 한다.

□ 경제분야별 우리나라의 세계 시장에서 차지하는 위상을 진단(순위, 점유율 등)하고, 특히 지식재산분야의 상대적으로 높은 순위와 점유율을 소개 □ 미국발 세계적 경제위기를 설명하고, 위기 속에서도 우리나라 경제의 희망적 측면을 설명 □ 경제위기 극복을 위한 녹색성장의 미래준비 o 녹색성장의 필요성을 사례를 통해 설명하고, 세계 주요국의 녹색성장 노력을 설명 o 녹색성장을 위한 국가간 경쟁에서 우위를 점하기 위한 수단으로서의 지식재산의 중요성을 설명하고, 각국의 지식재산 강화 정책을 소개 o 우리나라가 지식재산 전쟁에서 승리하기위한 조건들을 제시하는 동시에 현재 노력들을 소개

Deep eutectic solvents (DES’s) are mixtures of ammonium salts and hydrogen bond donors such as 1:2 choline chloride/urea or choline chloride/glycerol. DES’s are potential green solvents because they are nonvolatile, non-flammable, non-toxic and inexpensive. Many hydrolases are active in DES’s with rates comparable to those in toluene and up to five-fold higher than rates in typical ionic liquids. The glycerol component of the DES did not compete in the transesterification, indicating that its reactivity in ChCl:Gly is >200- fold lower than expected. Lipase from Candida antarctica was at least 20 times more stable in ChCl:U than in solutions containing the same concentrations of choline chloride and urea. We hypothesize that the hydrogen-bond network that forms between DES components decreases their reactivity. Addition of 10-25% ChCl:Gly to aqueous solutions enhanced the activity of esterases up to three-fold and of epoxide hydrolase up to 20-fold. Thus, these polar solvents are suitable candidate solvents or cosolvents for biocatalysis.

The olfactory system plays an important role in recognizing environmental conditions. Conventional electronic noses have been developed; however, they have limitations in sensitivity and specificity. On the contrary, the biological olfactory system has extremely high selectivity and sensitivity. Since olfactory receptor genes were identified and cloned, various researches on olfactory systems have been carried out, and the interest in olfaction research has been increasing due to its potential industrial applications. In the smelling process, the binding of specific odorants to the olfactory receptor proteins is the initiation step in odor recognition and triggers signal transduction in a cell. Functional expression of the olfactory receptors on the surface of culturable cells is very useful for application to an olfactory sensor. In this presentation, I am going to report a successful demonstration of human olfactory receptor-functionalized carbon nanotube- and conducting polymer nanotube-based FET sensors. We achieved highly-specific detection of odorant molecules down to femtomolar range sensitivity. The selectivity of this device is also extremely high. This device responds to the femtomolar concentrations of amylbutyrate; however, it does not produce any response to the related esters such as butyland hexyl butyrates that differ from the target compound by a single carbon atom each.

As our concerns on environmental problems are increasing, there has been much interest in developing sustainable system for the production of chemicals, fuels, and materials from renewable resources. Many microorganisms are capable of producing polyesters known as polyhydroxyalkanoates. However, microorganisms isolated from nature are often inefficient in performing the desired task. Thus, metabolic engineering has been employed for the improvement of microbial performance. In this lecture, I will describe the past, present, and the future of microbial polyesters, which will also cover design and production of natural polyesters to unnatural polyesters. Professor Ho Nam Chang’s great contribution to the field will also be described. [This work was supported by the Korean Systems Biology Research Grant from the MEST. Further supports by LG Chem Chair Professorship, BK21 program, and WCU program of the MEST are appreciated.]

Academia and research institutes have contributed to the development of Korean scientific technologies by innovative R&D performances. However, to create industrial value of these innovative R&D performances, there must be a commercialization process through technology transfer. In fact, due to inactive technology transaction market in Korea, these potent technologies are not even being subjected and just become hoarded or transferred with underestimated value. Thus, variety of strategies is needed to improve the effectiveness of technology transfer and to secure the true value. In order to create licensing, different strategies must be planned for each and every step of R&D performances, and according to the licensee’s capability. This presentation introduces domestic technology transfer consulting organs, and also it includes suggestions of effective technology strategies derived from the analysis of past successful technology transaction.

Molecular imaging technology for the development of new medicine is an important research field. Recently, as the cell-based assay technology is spreading quickly as a core technology for the discovery of new medicine, the development of high contents screening system, which has wide range of application, has been successfully acquired through medicinal research using cell-image analysis technology, nervous path analysis and etc. We have carried out the project of ‘Development of high-resolution HCS system equipped with micro optics and fluidic module’ as a subproject of ‘Development of High Content Screening Technology and System’ supported by the Ministry of Knowledge Economy. NEXMAN NH-600S from Nexstar Technology Co., Ltd. is true confocal fluorescent analysis system for high throughput cellbased imaging and screening. The system features 4 lasers (405nm, 488nm, 532nm and 635 nm), 4 PMTs for fluorescent detecting, a resonant scanner for X scanning and a galvanometer for Y scanning.

Drug delivery technologies have developed into innovative platforms that will make impacts for the pharmaceutical industry in every therapeutic category. Recently, the applications of drug delivery technologies are enormous, offering big opportunities to cure diseases more effectively than ever before. Drug delivery systems can advanced a product’s medicinal value by enhancing pharmacological improvement over conventional products on the market. And also, the right drug delivery system can raise a product’s market value by improving side-effects, tolerability and compliance issues. Drug delivery technologies have also used as a tool extending a product’s life cycle management with high profitability. Developing new and improved pharmaceuticals with drug delivery technologies has giving opportunity to pharmaceutical company’s key products which face patent expiration, for expending product life and sustaining value- added products. The advanced drug delivery technologies are including extended- release oral formulations, organ specific target, ,transdermal patches, topical creams, nasal spray, etc. For example, almost all therapeutic proteins are delivered by intravenous, subcutaneous, or intramuscular injection. A major reason for better protein delivery is needed is that patients do not like needles, especially when frequent injections are needed for treating chronic diseases. Fear and dislike of syringes is freighted with implications for compliance and health costs. Oral delivery is unquestionably the most popular form of drug delivery administration. Injection are probably the least. Today, bioavailability rarely reaches 1% when therapeutic peptides and proteins are taken orally. .If one can develop a new technology to have with over 10% bioavailability if oral protein delivery, it will be a great success for the market. The contents of this presentation will be reported the progress of “Development of controlled drug delivery system” supported by the Ministry of Knowledge and Economy (MKE) and 5 different companies for 5 years project. The program is composed by 5 subprojects as (1) development of novel osmotic pump, (2) liposome delivery system for anti tumor , (3) drug delivery system by bile acid transporter for oral delivery of antibiotics, (4)protein delivery of system using thermo-sensitive hydrogel, and (5) novel transdermal drug delivery system for anti-inflamation.

o 목표 - 천식치료제 톨로부테롤, 소염진통제 록소프로펜 등, 1 종 이상의 새로운 경피 투과 약물전달시스템 개발 - 유효 약물을 함유하는 다양한 약물전달 경피 투과시스템 개발에 의한 전임상, 임상시험 및 생산 적용 o 내용 - 소염진통제 록소프로펜의 피부투과 평가 - 록소프로펜의 최적 공정화 테스트 및 최적화 - 소염진통제 경피제제의 대량생산에 따른 재현성 및 함량균일성 달성 - 피부부착 후의 피부자극 등의 독성 평가 - 수동수송과 능동수송 병용가능한 담제 기술 축척 - GMP 공정에 따른 제조공정 확립

Molecular imaging technology is becoming more important in biotechnology research, articularly in the development of new drug compounds. High-content screening (HCS) is a drug screening and discovery technique that utilized high resolution fluorescence imaging of live cells. Protein targets of interest inside the cells are detected using fluorescent tags such as the green fluorescent protein or synthetic fluorescence dyes. As such, HCS can be defined as a ‘cell-based assay’ based on molecular imaging to evaluate the various biological functions of a drug candidate. For a successful development of a HCS system, various expertise such as molecular and cell biology, cellular and fluorescence imaging, automation and robotics, and image analysis software should be systematically integrated. From January of 2004, we have carried out the project of ‘Development of High Content Screening Technology and System’ supported by the Ministry of Knowledge Economy. The project consists of three (3) subprojects: (1) Development of high-resolution HCS system equipped with microoptics and fluidic module, (2) Development of image analysis software and system validation technologies, and (3) Development of HCS biocontents commercialization technology. Now, we have successfully developed a prototype of an HCS system, NEXMAN NH-600S. This system can be used for various applications including but are not limited to: drug candidate screening, toxicology testing, environmental risk assessment, etc. In this presentation, the fundamentals and results of the HCS development activities are discussed.

We previously demonstrated that the growth of Her-2/neu-overexpressing tumors could be effectively suppressed by the administration of plasmid DNA encoding a part of Her-2/neu (HM) and GM-CSF (pCK-HM-GMCSF) (Int. J. Cancer (2004) 111:86-95). The follow-up studies showed that the anti-Her-2/neu immune responses could be enhanced by combining adenovirus expressing HM (Ad-HM) with pCK-HM-GMCSF. To investigate the feasibility of this combined treatment in humans, its potency and safety were evaluated in large animals such as beagle dogs and cynomolgus monkeys. Beagle dogs were intramuscularly sequentially administered with pCK-HM-GMCSF and Ad-HM. The anti-Her-2/neu antibody titer was rapidly increased just after the last injection and reached the peak value 2 or 3 weeks post-injection. Repeated plasmid DNA injection could successfully boost humoral immune responses and maintain the high titer for more than one year. Furthermore, Her-2/neu-specific IFN-gamma production was detected in both peripheral blood mononuclear cells and splenocytes of immunized beagle dogs. In order to investigate which the combined treatment could break the immune tolerance, cynomolgus monkeys were administered as described above. The production of anti-Her2/neu antibodies, Herceptin-inhibition ability of the antibodies, and ADCC activities were demonstrated. In addition to humoral immune responses, the cellular immune responses were also analyzed using T cell proliferation assay, cytotoxic T lymphocyte assay, and IFN-gamma ELISA. All assays revealed that this gene-based cancer vaccine induced Her-2/neu-specific potent cellular immune activities (Gene Ther (2008) 15:1351-1360). In order to determine the safety of the combined treatment, we did a range of studies such as single-administration toxicity, repeated-administration toxicity, genotoxicity, and immunotoxicity. These studies did not show any material-related toxicity including autoimmune responses against host DNA, changes of lymphocytes population, hepatotoxicities, and cardiac toxicities. These results demonstrated both the efficacy and the safety of this combined treatment against Her-2/neu and strongly supported the feasibility of its use in humans.

Glycans are important components of many biotherapeutic agents, ranging from natural products to molecules based on rational design to recombinant glycoproteins1,2. The glycan components of these agents determine their biological activity and therapeutic efficacy. There are increasingly strong drivers for the development of improved approaches for production of therapeutics with enhanced efficacy and stability. The ultimate goal of our research project, supported by Korean Ministry of Knowledge Economy, is to develop next generation therapeutics through glycan remodeling based on glycomics analysis3, the systematic elucidation of the structure and function of “glycome (complete sets of glycans)” on bio-pharmaceuticals. The project was initially composed of five subprojects; (1) Subproject I: Development of anti-lymphoma therapeutic antibodies by glycoengineering (Mogam/Greencross), (2) Subproject II: Development of HBV therapeutic vaccines by glycan remodeling (Dobeel/Neurotech), (3) Subproject III: Development of therapeutic enzymes for fabry disease by glycan addition (ISU Abxis), (4) Subproject IV: Development of hyaluron derivatives by glycan modification (LG Life Sciences), and (5) Subproject V: Development of glycoexpression systems for glycan remodeling (KRIBB). The successful achievements of the project will surely upgrade the usage potential of these molecules, providing routes to better-targeted, more bioactive drugs with reduced off-target side effects and expanded applications.

Hepatitis B virus is a major cause of liver disease worldwide. Although currently available HBV vaccines are highly effective, HBV infection is still a major problem in Asia, Latin America, Africa and Eastern Europe. The WHO estimates that two billion people have been infected with HBV and about 350 million people have chronic infections, which can ultimately cause liver cirrhosis and cancer. When adults are infected with HBV, about 5-10% of them become chronically infected. However, vertical infections from mother and infections to neonates lead to chronic infections in almost 100% of cases. Additionally, greater than 90% of HBV infections in babies younger than 10 months result in chronic infection. Therefore, an improved HBV vaccine that can elicit protective immunity within 1-2 months would be beneficial, since currently available vaccines take 7-10 months to produce protective immunity. We have developed a CHO cell line that produces HBV surface antigens, including the L-protein, M-protein, and S-protein. The purified HBV surface antigens in the form of virus-like particles are highly immunogenic in mice, inducing more HBV-specific antibodies than any other vaccines on the market. This vaccine has been further improved by using an adjuvant that we have also developed, inducing strong immune responses in HBV transgenic mice, which then efficiently cleared HBV antigens in sera.

The aim of immunotherapy is to treat obstinate diseases, such as cancer, infectious diseases or autoimmune diseases by stimulating immune responses or suppressing harmful immune responses. Due to their selectivity and low toxicity, the immunotherapeutic agents become prominent as the next generation drugs. Furthermore, the therapeutic effect of chemotherapy or surgery is expected to be improved by combination with them. Here, we have developed 2 antibodies, 3 therapeutic vaccines and 2 cytokine/cytokine receptor. We produced HBV-neutralizing human monoclonal antibody and RSV-neutralizing human monoclonal antibody to prevent viral infection. Therapeutic vaccines for treatment of Her-2/neu overexpressing cancer, chronic HCV-infection or CEA-expressing solid cancer have developed and produced in accordance with good manufacturing practice. A new TNFR-Fc fusion protein for treatment of rheumatoid arthritis and a new G-CSF conjugate for treatment of neutrophenia were developed to improve in vivo stability and bioactivity.

MRC-01 (common name: methiozolin), 5-(2,6-difluoro-benzyloxymethyl)-5-methyl-3-(3-methyl- thiophen-2-yl)-4,5-dihydro-isoxazole, is a new herbicide candidate. This compound belongs to isoxazoline family, but not related to any other commercialized pesticides. MRC-01 has safety to various crops including soybean, wheat, rice, cotton, and turfgrass, and controls certain grass weeds such as barnyardgrass (Echinochloa crusgalli), crabgrass (Digitaria sp.), annual bluegrass (Poa annua), blackgrass (Alopecurus myosuroides) etc by pre or post-emergence application at 250 to 1,000 g/ha. In turf, MRC-01 has a high safety to various cool season turf species including bentgrass and Kentucky bluegrass, and controls the major grass weeds (annual bluegrass and crabgrass). MRC-01 is superior to the commercial standards in post emergence activity. MRC-01 mode of action was shown, by [14C]-glucose incorporation study, to be a strong inhibitory activity in cell wall biosynthesis in maize. Toxicology studies have proven that MRC-01 has a high safety to mammals, fishes, bees, earthworm, and birds etc. The molecule was non-genotoxic and nonteratogenic. Animal metabolism study showed that MRC-01 was readily absorbed in the gastrointestinal tract, rapidly metabolized in the liver, and excreted through bile, faces, and urine. Greater than 90% of the administered was excreted out within 96 h. In rats, numerous minor metabolites were formed and one of the metabolites was identified to be 2-(5-((2,6- difluorobenzyloxy)methyl)-4,5-dihydro-5- methylisoxazol-3-yl)thiophen-3-yl)methanol. In soil, MRC-01 was readily degraded in a non-sterile aerobic condition. In field dissipation studies, DT50 of MRC-01 was about 10 days. Collectively, MRC-01 has a unique fitness as a turf herbicide and favorable toxicology and environmental fate. MRC-01 was applied for registration for turf uses in September 2009 to KRDA, and is in registration trials in Japan.

Bone marrow stromal cells or MSCs exists as a population of cells having the capacity to differentiate into osteogenic, chondrogenic or adipogenic lineages in vitro and in vivo. To utilize the enormous MSCs potential for cartilage injury repair, numerous attempts have been made, including MSC-seeded scaffold and cultured MSCs implantation in vivo. Formation of biological substitute using the concept of tissue engineering generally requires an artificial scaffold. The three dimensional environment plays an important role in promoting cell-matrix interactions during chondrogenesis and the phenotypic stability of chondrocytes are closely related to their surrounding 3D matrix. The extracellular matrix (ECM) has received significant attention as a functional scaffold in the tissue engineering field, because it can serve not only a physical support but also provide favorable environments for cells. However, the ECM scaffold still requires many technical improvements to construct artificial tissue suitable for clinical use. So, we developed a novel ECM scaffold fabricated by our proprietary procedure using cultured chondrocyte and focus on ECM application to clinical use. Until now, we apply that novel ECM scaffold and membrane were developed. And, we demonstrated that ECM would confer benefit as a result of cartilage defect after ECM scaffold or membrane implantation.

o 목표 - 세포치료제 생산의 효능 증대를 최대화 할 수 있는 세포치료제 특이 자동 배양 시스템 개발 및 제품화 o 내용 - 세포치료용 성체줄기세포 제조 공정 표준화 - 동물 모델을 이용한 안정성 및 유효성 검증 - GMP 시설 내 임상적용 줄기세포의 안정적 생산 - 신경손상질환 세포치료제 IND 신청 - 무혈청 배지 적응 및 배양 공정법 개발

본 과제는 5개의 세부과제로 구성되어 있으며, 최종목표는 새로운 작용점 1개 이상 발굴 및 신규 작물보호 생리활성 상업화 물질2개 이상을 선발하는 것이다 제노마인(주)과 화학연구원이 공동으로 연구중인 작용점 발굴연구는 현재 신규 제초제 작용점으로 AtKAPAS, AtCAD, AtPDX4, AtPDX5, AtSNO와 AtMSG, 신규 살충제 작용점으로 trehalase, 그리고 신규 살균제작용점으로 melanin 생합성 효소 등 총 8종의 신규작용점 후보를 발굴하였으며, 신규 작용점 발굴부터 제초제 후보물질 탐색 및 개발에 대한 시스템을 확립(TargetScrTM)하였다. 특히, KAPAS는 본 시스템을 적용하여 제초활성을 지닌 선도물질 KHG23844(특허 출원 10-2008-0046112)를 선발하였다. 신규제초제 개발 연구는 (주)목우연 구소에서 신물질 밭제초제 methiozoline(MRC-01)을 선발하였는데, 신규 작용기작으로 잔디에 적용성이 탁월하여 상업화 개발이 진행 중이다. 원제이화학성, 제제 이화학성, 인축 독성, 환경 독성, 토양 잔류, 동물 대사, 약효. 약해 시험 등 국내등록자료의 확보 및 등록 신청을 완료하였고 일본에서 개발 중이며 골프장 그린에 발생하는 새포아풀에 탁월한 효과를 선택적으로 보이며 바랭이에도 효과가 있다. 한편, (주)동부하이텍에서는 고활성이며 친환경적인 비선택성 제초제 DCC-3825 를 상업화 물질로 선발하였다. Paraquat 대체와 Glyphosate 저항성 잡초 방제를 목표로 하고 있으며, 특히 화본과 식물 및 클로버에 탁월한 효과를 갖고 있다. 예비 안전성시험을 완료하였고 등록을 위한 독성시험 등을 수행할 예정이다. 신규살균제 개발 연구는 (주)경농에서 Broad Spectrum을 갖는 살균제 KNF05234 선발하여 기존 strobilurin계통의 약점인 약해를 줄이고 저항성 병균에 탁월한 효과가 있음을 확인하여 현재 글로벌 원제 등록을 위한 CRO 계약을 완료하였고, 현재 38항목에 대한 인축독성 시험을 국내외에서 진행중이다. 동부정밀화학(주)는 살충제 후보물질 도출을 목표로 하여 1단계에 octopamine receptor 저해제로서 응애류를 방제할 수 있는 신규살충제 후보물질 K16776을 발굴하고 포장실험을 진행하였다. 2단계에서는 나방류의 유충에 특히 활성이 높은 ryanodine receptor agonist에 관한 연구를 진행하여 대조 화합물과 유사한 정도의 살충활성을 지닌K17704, K17710등의 선도화합물을 확보하고, 문제점 개선을 위한 구조적 최적화가 진행중이다. 본 과제의 성과 달성도는 100%이며, 과제 성과물을 통한 경제적 효과는 작용점 1개의 매출 총액 11.6억불/년, 잔디용 제초제 (국내 150억원, 국외 5.2억불), 광범위 살균제 16.6억 불, 비선택성 제초제(국내 250억원, 국외 2.25천만불)를 포함하여 총 약 3조 4,000억원에 이를 것으로 예상된다.

Asthma is one of the major public health problems worldwide and the morbidity and mortality of asthma has increased in the past two decades. To develop the botanical new drug for asthma, we discovered natural product that activated beta2-adrenergic receptor(　2AR) in the reporter assay. It also inhibited nuclear factor (NF)-　B activation. Administration of the NDR10113 to ovalbumin (OVA)-induced asthmatic mice improved the airway hyperresponsiveness (AHR). Moreover, NDR10113 exhibited a decrease of serum IgE level and bronchial inflammation. Results from flow cytometric analysis (FACS) showed the absolute number of total cells in the lung-draining mediastinal lymph nodes (MLN) were significantly suppressed by NDR10113 administration, and also that absolute numbers of CD3+ T cells, CD4+ Th cells, and B220+ B cells reduced. Furthermore, the lung tissue of NDR10113 treatment group showed much less eosinophils, bloods, and collagen accumulating compared with that of OVA-induced control group. Taken together, these results suggest NDR10113 might be beneficial for the treatment of asthma.

The pharmaceutical industry has traditionally focused on developing therapies using synthetic small molecules. However many environments such as a generic threat and an R&D productivity crisis have changed the dynamics of the pharmaceutical industry. The major players in the industry have recognized the high potential of biological therapies. In near future, it is expected to drive a significant change in their R&D portfolios. Biologics are expected to account for the majority of the total growth in the pharmaceutical sector. In 2014, seven among the top 10 products by sales will be the Biologics. One of the major fields for application of biological products is the autoimmune disease such as a Rheumatoid arthritis. The market size of the TNF blockade will be expended to 20 billion dollars. The final goal of our program is to commercialize the biologics as a best in class drug and/or as a first in class drug for the treatment of autoimmune diseases.

o 목표 : 유용 리간드 발굴 및 응용을 통한 의약 후보 물질 발굴 o 내용 리간드를 이용한 난치성 질환에 대한 효과적 치료제가 급증하고 있는 시점에서, 생체 활성 리간드의 범위를 확장하고, 치료제 개발을 위한 유용 리간드의 발굴을 융합 기술 기반을 구축하는 것은 학술적으로나 경제적으로 그 가치가 크다고 할 수 있다. 이를 위해 과제 1단계에서는 유용 리간드 발굴을 위한 융합 기술 및 인프라를 구축하여 2단계에서 유용 리간드의 약효 검증 및 효능 평가를 통한 전임상 후보 물질을 개발해 왔고, 3단계에서는 최종적으로 임상 후보 물질 발굴을 목표로 하고 있다. 각 세부 과제별로 1세부 과제에서는 줄기세포의 골 분화를 유도하는 신규 기전의 골다공증 치료제 개발, 2세부 과제에서는 신개념 항균 소재를 개발, 3세부 과제에서는 천연물 유래 천식 치료제 개발을 목적으로 하고 있다. 이러한 과제를 수행함으로써 기술적 측면에서 핵심 기반기술 및 연구 인력 확보를 통해 세계 경쟁력을 증진시키고, 지속적인 유용 리간드 발굴이 가능하며, 뿐만 아니라 경제·산업적 측면에서 지적재산권, 기술 이전 및 로열티 확보, 치료제의 산업화를 통한 수익 창출을 기대할 수 있다.

A p40-hyFc is a Fc fusion protein consisting of human p40 (a member of IL-12 family) and hybrid Fc (hyFc) derived from human IgD and IgG4 Fc regions. The p40 has been well known as a natural antagonist of IL-12 and IL-23 (1). Recently, it was reported that Th17 cell is a main regulator for autoimmune diseases and expanded by IL-23 (2), suggesting that the blockage of IL-23 signaling by long-acting p40 protein could be new approach to treat autoimmune diseases. The human p40-hyFc was demonstrated to specifically block IL-23 signaling in PBMCs from rheumatoid arthritis patients and mouse p40-hyFc (derivative of human p40-hyFc) also inhibited IL-23-mediated signaling in vitro and could decrease arthritis index score in mice collagen-induced arthritis (CIA) model In PK analysis using three cynomolgus monkeys, single subcutaneous injection with human p40-hyFc showed long half-life in monkeys. There was no significant increase in CBC and lymphocytes and no detectable Ab responses against human p40-hyFc. In cross-reactivity test, the human p40-hyFc did not show cross-reactivity with rodent, but did with monkeys. Therefore, the rhesus monkey model of CIA is the only available option to evaluate the efficacy of IL-12p40-hyFc in vivo. At present, the efficacy of human p40-hyFc was being evaluated in monkey CIA model in Nertherland primate center. To perform preclinical study, a stable cell line expressing human p40-hyFc was established, then MCB/WCB manufacturing and their characterizations are being conducted in Apptec company in USA. The process development for cultivation, purification, and analysis for p40-hyFc has been performed during this study. Since there are non-responders to commercially available drugs such as Enbrel (TNFR-Fc) and Orencia (CTLA-4-Fc) targeting to soluble TNF- and CTLA4, respectively, human p40-hyFc could be a novel drug with different action mechanism and be used in combination with these therapeutics for patients.

A new strategy for highly sensitive and rapid protease assay is developed by mediating proteolytic formation of oligonucleotide duplexes and using the duplexes for signal amplification. In the presence of matrix metalloprotease-2 (MMP-2), fragmentation of the intact DNA-peptide on gold nanoparticles (GNP) by hydrolytic cleavage of a peptide bond within the substrate allows diffusion of DNA away from GNP and the formation of a DNA/RNA heteroduplex, leading to digestion of RNA by RNase H. Because of the high quenching efficacy of GNP to the fluorophore in RNA and multiple digestions of the RNA, the fluorescence signal recovery is amplified. This method permits the assessment of the activity of MMP-2 at concentrations as low as 10 pM within 4 hours. Compared with the reported protease nanosensors using quantum dots, GNP, and magnetic nanoparticles with the same peptide sequence, the assay time of this method is 6-fold faster and the limit of detection is 100-fold more sensitive. We believe that the formulations for proteolytic formations of oligonucleotides duplexes for signal amplification on GNP will lead to the development of more sensitive and rapid protease assay techniques, thus extend the role of proteases as therapeutic targets and disease indicators

Many clinical conditions in neurosurgery, orthopedics, and dental surgery require bone regeneration. The indications include bone fracture, trauma, spinal fusion, and dental implantation. Therapies for bone regeneration are various. Some of them are being used clinically and some are currently under preclinical or clinical studies. The therapies include bone grafts, bone morphogenetic protein-2 (BMP-2) protein delivery, stem cell transplantation, and gene delivery. Calcium phosphatebased ceramics, such as hydroxyapatite (HA) and tricalcium phosphate, have been used as bone substitutes, but these materials have poor mechanical performance. Recently, increasing interest has focused on using polymer/ceramic composite materials as bone substitutes to improve mechanical properties and control the biodegradation rate easily [1]. Polymer/ceramic composites have a non-uniform and limited exposure of the ceramics on the scaffolding surface. This is the result of the polymer solution enveloping the ceramic particles during the fabrication process. This problem can be overcome by two methods. One is fabricating polymer/ceramic composites using the gas-foaming method [1]. The other is exposing nano HA particles on the polymer/HA scaffolds acting as nucleation sites for apatite deposition and accelerating the apatite growth rate in a biomimetic coating approach using a simulated body fluid [2]. BMP-2 is one of the most potent growth factors for the induction of bone formation in vivo. The bone formation efficacy of BMP-2 depends on the protein delivery system, because BMPs administered in solution may lose their bioactivity over a short time and do not always exhibit the required efficacy in bone regeneration in vivo. We have developed BMP-2 delivery systems that allow for sustained and localized release of BMP-2 protein by conjugating heparin, a polysaccharide macromolecule that binds directly with BMP-2 and helps to retain BMP-2 activity, to polymer scaffolds [3], gels, or nanoparticles [4]. Stem cell transplantation can also induce bone regeneration. Generally, bone formation by stem cell transplantation requires osteogenic differentiation of the cells prior to transplantation. However, the in vitro osteogenic differentiation requires additional culture periods of 3-4 weeks, during which patients should wait. Recently, we have developed a technology in which stem cells that are not differentiated osteogenically in vitro prior to transplantation extensively regenerate bone in vivo when exogenous BMP-2 is delivered to the transplantation site [5,6]. This method does not require an additional culture period for in vitro osteogenic differentiation and would be valuable for bone regeneration.