• 대한의생명과학회지
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• 제9권2호
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• pp.67-74
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• 2003

Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

• KSBB Journal
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• 제15권5호
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• pp.497-500
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• 2000

Avidin의 금속표변에 대한 강한 결합력과 avidin-biotin의 강한 결합력을 이용하여 금속 표면 위에 라포좀과 같은 유기 분자막의 다층 형성 과정을 수정진동자를 이용하여 분석하였다. 금속 표변위에 적층되는 유기 분자막에 대한 정보를 수 집하고 그 가능성을 검토하여 바이오 센서에서 감도를 향상 시킬 수 있는 새로운 방법을 제시하고자 하였다.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1209-1216
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• 2022

To better understand the effects of PEGylation and biotinylation on the delivery efficiency of proteins, the cationic protein lysozyme (LZ) and anionic protein bovine serum albumin (BSA) were chemically conjugated with poly(ethylene glycol) (PEG) and biotin-PEG to primary amine groups of proteins using N-hydroxysuccinimide reactions. Four types of protein conjugates were successfully prepared: PEGylated LZ (PEG-LZ), PEGylated BSA (PEG-BSA), biotin-PEG-conjugated LZ (Bio-PEG-LZ), and biotin-PEG-conjugated BSA (Bio-PEG-BSA). PEG-LZ and Bio-PEG-LZ exhibited a lower intracellular uptake than that of LZ in A549 human lung cancer cells (in a two-dimensional culture). However, Bio-PEG-BSA showed significantly improved intracellular delivery as compared to that of PEG-BSA and BSA, probably because of favorable interactions with cells via biotin receptors. For A549/fibroblast coculture spheroids, PEG-LZ and PEG-BSA exhibited significantly decreased tissue penetration as compared with that of unmodified proteins. However, Bio-PEG-BSA showed tissue penetration comparable to that of unmodified BSA. In addition, citraconlyated LZ (Cit-LZ) showed reduced spheroid penetration as compared to that of LZ, probably owing to a decrease in protein charge. Taken together, chemical conjugation of targeting ligands-PEG to anionic proteins could be a promising strategy to improve intracellular delivery and in vivo activity, whereas modifications of cationic proteins should be more delicately designed.

• 대한수의학회지
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• 제30권4호
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• pp.435-440
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• 1990

오제스키병바이러스 또는 돼지콜레라바이러스에 인공 또는 자연감염된 돼지 10두를 실험동물로 공시하였으며, 감염돈의 편도선, 비장, 대뇌 및 연층(buffy coat)의 냉동 및 파라핀절편에서, avidin-biotin-peroxidase complex(ABC)를 이용하여 이들 바이러스를 면역조직화학적으로 검출하였다. 오제스키병바이러스 항원은 임파구와 대식세포의 핵내 또는 세포질에서 검출되었으며, 돼지콜레라바이러스 항원은 이들 세포의 세포질에서 검출되었고, ABC법은 양성반응 부위에서 갈색의 색소 침착을 일으켰다. 오제스키병바이러스 양성세포는 편도선과 대뇌에서 가장 빈번히 검출되었음에 반해 돼지콜레라바이러스는 비장에서 가장 빈번하였다. 그리고 연충에서도 이들 두가지 바이러스항원이 모두 검출되었다. ABC법은 이을 바이러스의 면역조직화학적 검출에 있어 특이성이 높고 배경의 비특이염색성이 낮아, 바이러스 분리동정을 거치지않고 이들 질병을 확진할 수 있는 진단수단으로 활용될 수 있을 것으로 생각된다.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.601-610
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• 2003

Biotin is a water-soluble vitamin used as a skin conditioning agent and promotes the formation of intercellular lipid layers through increased lipid synthesis, which improves the skin's natural barrier function. The anti-inflammatory effects of biotin have been investigated using in vitro assay models, such as MTT assay, measurements of concentrations of nitric oxide(NO), prostaglandin E2(PGE$_2$), and inhibition rate of 5-lipoxygenase(5-LOX). In comparison with biotin, other plant extracts were tested at the same time which were kudzu vine extract, sage extract, paeonia extract, and dipotassium glycyrrhetinate. Nitric oxide is a signal molecule with functions such as neurotransmission, local vascular relaxation, and anti-inflammation in many physiological and pathological processes. NO can cause apoptosis and necrosis of target cells such as keratinocytes and is generated from L-arginine by nitric oxide synthase (NOS). Prostanoids, including prostaglandins and thromboxanes, are generated by the phospholipase $A_2$/cyclooxygenase(COX) pathway, and leukotrienes are generated by the 5-lipoxygenase pathway from arachidonic acid. Prostaglandin E2 recently have been shown to be beneficial in the resolution of tissue injury and inflammation, also has been implicated as an immunosuppressive agent and plasma levels of PGE$_2$ are elevated in patients sustaining thermal injury. Lipoxygenase metabolites from arachidonic acid have been implicated in inflammation, anti-inflammatory activity of the raw materials was evaluated in vitro by the offered inhibition of lipoxygenase.

• 미생물학회지
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• 제54권4호
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• pp.330-342
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• 2018

Streptavidin (STR)과 Biotin system은 Biotin의 Streptavidin에 대한 높은 비공유 친화력(non-covalent affinity; $K_D=10^{-14}M$)과 4 Biotin 결합부위를 갖는 Streptavidin의 tetramer 구조로 인해 복수의 항원결합부위 및 복수의 항원특이성을 갖는 항체를 제조할 수 있기 때문에 가장 활발하게 연구되고 있다. 이 system을 활용하기 위해 우리는 Streptomyces avidinii 염색체 DNA로부터 PCR을 통해 Streptavidin (STR) 유전자를 증폭하고 이를 TRAIL (tumor necrosis factor ${\alpha}$ related apoptosis induced ligand) receptor인 death receptor 4 (DR4)에 특이적으로 결합하는 hAY4 single-chain Fv 항체유전자에 융합시켰다. 대장균에서 발현시킨 STR에 융합된 hAY4 ScFv (hAY4-STR) 항체는 가열시킨 SDS-PAGE에서 43 kDa monomer를 나타내었다. 그러나 가열하지 않은 SDS-PAGE와 Size-exclusion chromatography에서는 tetramer인 172 kDa을 나타내었는데 이는 hAY4 ScFv-STR 항체가 STR의 자연적인 비공유결합에 의해 유도된 tetramer를 형성하고 있음을 나타내고 있다. 본 융합 단백질은 Ouchterlony assay와 ELISA에서 보여주는 것처럼 자연 Streptavidin과 유사한 Biotin 결합력을 유지하고 있었다. ELISA와 Westernblot을 이용하여 정제된 hAY4-STR 융합항체의 DR4 항원결합력 또한 확인하였다. 게다가 표면 플라즈몬 공명(surface plasmon resonance) 분석에서 hAY4 ScFv-STR tetramer는 tetramerization에 의해 hAY4 ScFv monomer보다 60배 더 높은 항원결합력을 나타내었다. 요약하면 hAY4 ScFv-STR 융합단백질은 E. coli에서 soluble tetramer로 성공적으로 발현 및 정제되었으며 Biotin과 DR4 항원에 동시에 결합함을 보여 주었다. 이는 bifunctional and tetrameric ScFv 항체를 제조 할 수 있음을 제시해 주고 있다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.546-549
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• 2000

Avidin-biotin의 강한 결합력을 이용하여 금속 표면 위에 리포좀과 같은 유기 분자막의 다층 형성 과정을 QCA의 공진주파수와 공진저항의 변화를 측정하므로서 실시간 모니터링의 가능성을 검토하고, 유기 분자막이 금속 표면 위에 적층 됨에 따라 형성되는 적층 막에 대한 정보를 수집함으로서 바이오센서 시스템으로 QCA를 적용 가능함의 기초 데이터를 제공하고자 한다.

• 센서학회지
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• 제4권4호
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• pp.81-87
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• 1995

A novel biomaterial capable of incorporating biotinylated biomolecule has been synthesized. Our strategy is to biotinylate one-dimensional electroactive polymers and use a bridging streptavidin protein on Langmuir-Blodgett (LB) organized films. These copolymers are derivatized with long alkyl chains and biotin moieties to bind, respectively, to the hydrophobic surface and the biotinylated species, through the biotin and streptavidin complexation. We utilize the polymer assembly approach to attach a signal transducing biomolecule biotinylated phycoerythrin (B-PE) into this novel biomaterial by binding the unoccupied biotin binding sites on the bound streptavidin (4 sites total). The pressure-area isotherm of the protein injected monolayer showed area expansion. A characteristic fluorescent emission peak at 576nm was detected from the monolayer transferred onto a solid substrate. These observations demonstrated the promise of the organized thin polymer assemblies for their application to the sensor system.

• 한국CDE학회논문집
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• 제12권4호
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• pp.298-307
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• 2007

In recent years, researchers are actively investigating new methods that are applicable for manufacturing micrometer to nanometer scale structures. Among them, optical tweezers that can manipulate microscopic objects using a laser is receiving one of the key attentions. Optical tweezers have been used actively in the field of science. For example, for measuring mechanical characteristics in the scale of piconewtons or for manipulating and sorting large numbers of particles, bacteria, cells. etc. However, little works have been reported for "manufacturing" objects. In this paper, we present a new method for manufacturing micrometer scale structures using micrometer scale biotin coated polystyrene particles. Particles will be controlled with a user interface that utilizes a CyberGlove and glued together by the bonding force between biotin and streptavidin.

• Bulletin of the Korean Chemical Society
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• 제17권11호
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• pp.1018-1022
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• 1996

Adenosine deaminase (ADA) has been utilized as the label in devising a potentiometric homogeneous assay for riboflavin and riboflavin binding protein (RBP). The proposed homogeneous assay method employs an ADA-biotin conjugate as the signal generator and an avidin-riboflavin conjugate as the signal modulator in the solution phase. The catalytic activity of the ADA-biotin conjugate is inhibited in the presence of an excess amount of the avidin-riboflavin conjugate, and the observed inhibition is reversed in an amount proportional to the concentration of RBP added. When the analyte riboflavin is added to this mixture of ADA-biotin, avidin-riboflavin and RBP, the activity of the enzyme conjugate is re-inhibited in an amount proportional to the concentration of riboflavin. Since the enzyme label used in this system is ADA, an ammonia-producing enzyme, a potentiometric rather than photometric detection scheme is used to monitor the enzymatic activity in the assay.