• 대한의생명과학회지
• /
• 제10권3호
• /
• pp.211-217
• /
• 2004

For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

• 사물인터넷융복합논문지
• /
• 제4권2호
• /
• pp.13-20
• /
• 2018

본 논문은 최근 대두된 신기술인 '블록체인' 기술을 기반으로 'Single-Sign-On'과 'Token 기반 인증 방식'을 접목한 인증 시스템을 제안하였다. Single-Sign-On 기반 인증 방식에 블록체인 기술을 접목하여 '접근제어' 기능과 '무결성'을 제공하였으며, Token 기반 인증 방식을 사용하여 Stateless한 Self-Contained 인증 기능을 제공하였다. 암호화 기반 Token 발급 및 인증 과정을 수행하여 보안성을 높일 수 있었으며, Web Server에 대한 인증 편리성을 제공하였다. 또한 SSO과 Token 기반 인증을 통해 번거로운 인증 과정을 보다 편리하게 개선할 수 있는 방법을 제시하였다.

• 품질경영학회지
• /
• 제28권4호
• /
• pp.204-222
• /
• 2000

Supply Chain Management is a process-oriented concept, integrated approach to procuring, producing, and delivering products and services to customers. Supply Chain Management covers the management of material, information, and funds flows. A strategic supply chain management produces for value maker and the ultimate customer. One aim of this paper is to determine the effect of contract parameters on the quality of the end product of the supplier-producer chain. The supply chain management is viewed as a single entity rather fragmented. This research constructed model the relationship of supply chain management to quality management. The questionnaire were tested with survey of supplier's employee and the result were analyzed using SEM(Structural Equation Model). As a result, Quality performance positively depends on the communication, coordination, and process management between the supplier and the producer.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권4호
• /
• pp.498-503
• /
• 2003

Luteinizing hormone as other glycoprotein hormones is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a specific $\beta$-subunit. The correct conformation of the heterodimer is important for efficient secretion, hormonal-specific post-translational modifications, receptor binding and signal transduction. To determine whether $\alpha$- and $\beta$- subunits can be synthesized as a single polypeptide chain (tethered-bLH and -bFSH) and also display biological activities, the tetheredbLH and -bFSH molecules were constructed and transfected into chinese hamster ovary (CHO-K1) cells. LH and FSH activities were assayed by using the human embryonic kidney (HEK) 293 cells expressing rat LH and FSH receptor genes. The tethered-bLH and - bFSH proteins were efficiently secreted and showed a similar activity to the dimeric bovine LH and FSH $\alpha$/$\beta$ wild type and native purified from bovine pituitary. The tethered-molecules can be permit development of potent new analogues that stimulate ovarian development. Taken together, a single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds. These data indicate the potentiality of the single chain approach to further investigate structurefunction relationships of LH and FSH.

• 대한교통학회지
• /
• 제29권3호
• /
• pp.7-19
• /
• 2011

본 연구에서는 통행사슬(Trip Chain)개념을 이용하여 개인이 통행시 통행목적과 통행연계수를 고려하는 행태에 대하여 분석하였다. 통행사슬을 먼저 통근통행을 포함하는 통행과 통근통행을 포함하지 않는 기타통행으로 구분하고 각각의 통행사슬에서 한 곳의 경유지만을 이동하는 단순연계통행과 2개 이상을 경유하는 복합연계통행으로 구분하였다. 방법론으로는 2단계 네스티드 로짓모델(Nested logit model)을 이용하여 분석하였으며 그 결과, 출생년도가 높을수록 남성일수록, 고정사무실기반 직장일수록 단순연계통행보다는 복합연계통행을 할 경향이 높게 나타났다. 반대로 차량을 보유할수록, 월수입이 300만원미만일수록, 평균통행시간과 평균체류시간이 오래 걸린 응답자일수록 단순연계통행일 경향이 높은 것으로 분석되었다.(p<0.001) 본 연구에서는 2006년 수도권 가구통행실태 조사자료를 이용하였다.

• IMMUNE NETWORK
• /
• 제12권1호
• /
• pp.33-39
• /
• 2012

Background: Therapeutic approaches using monoclonal antibodies (mAbs) against complement regulatory proteins (CRPs:i.e.,CD46,CD55 and CD59) have been reported for adjuvant cancer therapy. In this study, we generated a recombinant 1E8 single-chain anti-CD59 antibody (scFv-Fc) and tested anti-cancer effect.by using complement dependent cytotoxicity (CDC). Methods: We isolated mRNA from 1E8 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reversetranscriptase polymerase chain reaction (RT-PCR). Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. The VL and VH fragments were cloned into the pOptiVEC-TOPO vector that contained the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1E8 single-chain Fv-Fc (scFv-Fc) constructs. CD59 expression was evaluated in the prostate cancer cell lines using flow cytometry. The enhancement of CDC effect by mouse 1E8 and 1E8 scFv-Fc were evaluated using a cytotoxicity assay. Results: The scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was similar to that of the mouse 1E8 for CCRF-CEM cells. The molecular masses of 1E8 scFv-Fc were about 120 kDa and 55 kDa under reducing and non-reducing conditions, respectively. The DNA sequence of 1E8 scFv-Fc was obtained and presented. CD59 was highly expressed by the prostate cancer cell line. The recombinant 1E8 scFv-Fc mAb revealed significantly enhanced CDC effect similar with mouse 1E8 for prostate cancer cells. Conclusion: A 1E8 scFv-Fc construct for adjuvant cancer therapy was developed.

• 멤브레인
• /
• 제12권4호
• /
• pp.207-215
• /
• 2002

한정된 미세공간에서의 제한확산(hindered diffusion)은 멤브레인 기공(pore)에서 입자들의 운동에 의해 결정되는 여과 메카니즘을 매우 미세한 수준에서 이해하는데 중요한 현상이다. 구형(spherical) 콜로이드 입자에 비해 보다 복잡한 형태(conformation)인 고분자사슬 구조를 갖는 다가전해질(polyelectrolyte)의 제한확산 거동에는 다양한 인자들이 관련되어 있기 때문에, 이론 접근은 물론 실험적 접근도 한층 어려운 것이 사실이다. 본 연구에서는, 슬릿형 미세기공에 한정되어 있는 단일한 다가전해질(single polyelectrolyte)에 coarse-grained bead spring model과 먼거리(long-range) 정전상호작용(electrostatic interaction)인 Debye-Huckel potential을 적용하여 분자시뮬레이션 기법인 브라운 동력학 모사를 수행하였다. 기공과 다가전해질 사슬(Polyelectrolyte chain)의 주어진 크기에서, 용액의 전해질 이온농도가 감소함에 따른 사슬의 신장(extension)효과는 제한확산계수를 감소시켰고, 기공 벽면의 하전성은 제한확산계수를 더욱 감소시켰다. 이는, 다가전해질 사슬(polyelectrolyte chain)의 입체적 장애(steric hindrance)와 함께 정전반발력이 미세기공에서의 확산이동을 억제함을 의미한다.

• Journal of Microbiology and Biotechnology
• /
• 제26권12호
• /
• pp.2184-2191
• /
• 2016

The time-consuming and high-cost preparation of soluble peptide-major histocompatibility complexes (pMHC) currently limits their wide uses in monitoring antigen-specific T cells. The single-chain trimer (SCT) of peptide-${\beta}2m$-MHC class I heavy chain was developed as an alternative strategy, but its gene fusion is hindered in many cases owing to the incompatibility between the multiple restriction enzymes and the restriction endonuclease sites of plasmid vectors. In this study, overlap extension PCR and one-step cloning were adopted to overcome this restriction. The SCT gene of the $OVA_{257-264}$ peptide-$(GS_4)_3-{\beta}2m-(GS_4)_4-H-2K^b$ heavy chain was constructed and inserted into plasmid pET28a by overlap extension PCR and one-step cloning, without the requirement of restriction enzymes. The SCT protein was expressed in Escherichia coli, and then purified and refolded. The resulting $H-2K^b/OVA_{257-264}$ complex showed the correct structural conformation and capability to bind with $OVA_{257-264}$-specific T-cell receptor. The overlap extension PCR and one-step cloning ensure the construction of single-chain MHC class I molecules associated with random epitopes, and will facilitate the preparation of soluble pMHC multimers.

• 대한산업공학회지
• /
• 제32권4호
• /
• pp.382-392
• /
• 2006

Vendor-managed inventory policy(VMIP) is a supply-chain initiative where the supplier is authorized to manage inventories of items at retail locations. In VMIP, the supplier monitors sales and stock information at retail locations and makes decisions of inventory replenishment and transportation simultaneously. VMIP has been known as an effective supply chain strategy that can realize many of benefits obtainable only in a fully integrated supply chain. However, VMIP does not always lead to lower the supply chain cost. It sometimes generates the total supply chain cost higher than the traditional retailer-managed inventory policy (RMIP). In this paper, we perform a comparison study on RMIP and VMIP in the retail supply chain which consists of a single supplier and a number of retailers. We formulate mixed integer programming models for both RMIP and VMIP with vehicle routing problems and perform computational experiments on various test problems. Furthermore, we derive the conditions which guarantee the dominant position for VMIP with respect to total supply chain cost in the simple retail supply chain.

• BMB Reports
• /
• 제40권6호
• /
• pp.875-880
• /
• 2007

Glucose oxidase (GOD) is an oxidoreductase catalyzing the reaction of glucose and oxygen to peroxide and gluconolacton (EC 1.1.3.4.). GOD is a widely used enzyme in biotechnology. Therefore the production of monoclonal antibodies and antibody fragments to GOD are of interest in bioanalytics and even tumor therapy. We describe here the generation of a panel of monoclonal antibodies to native and heat inactivated GOD. One of the hybridomas, E13BC8, was used for cloning of a single chain antibody(scFv). This scFv was expressed in Escherichia coli XL1-blue with the help of the vector system pOPE101. The scFv was isolated from the periplasmic fraction and detected by western blotting. It reacts specifically with soluble active GOD but does not recognize denatured GOD adsorbed to the solid phase. The same binding properties were also found for the monoclonal antibody E13BC8.