• 생명과학회지
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• 제28권12호
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• pp.1416-1423
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• 2018

Biotin에 강한 결합 친화력($K_D=10^{-14}M$)과 함께 streptavidin의 tetramer 특징은 VHH 항체를 streptavidin에 융합시키게 하여 biotinylated horseradish peroxidase를 사용하는ELISA 와Western blot analysis 등의 면역분석법에서 VHH 항체의 항원결합력을 증가시키는데 응용 가능하다. 이를 응용하기 위해 우리는 Streptomyces avidinii 염색체 DNA로부터 PCR을 통해 streptavidin유전자를 증폭하고 이를 green fluorescent protein항원에 특이적으로 결합하는 8B9 VHH 항체유전자에 융합시켰다. 대장균에서 수용성 융합단백질로 발현시키기 위해 pUC119 플라스미드에 기초한 발현시스템을 사용하였다. 즉 lacZ promoter를 사용하여 IPTG에 의해 단백질발현을 유도하게 하였으며, 아미노말단에 pelB leader를 두어 발현된 단백질의 periplasmic space로 이동하게 하여 수용성 단백질형태의 분비를 촉진하였으며 카르복시말단에 6개의 polyhistidine tags를 두어 $Ni^+$-NTA-agarose column을 사용하여 발현된 단백질을 정제하였다. Streptavidin이 biotin에 강하게 결합함으로 대장균에 독성을 나타냄에도 불구하고 본 수용성 융합단백질은 성공적으로 발현되었다. SDS-PAGE에서 가열하는 경우 변성되어 30.6 kDa를, 가열하지 않는 경우에는 자연 상태의 122.4 kDa을 나타내었다. 이는 8B9 VHH항체에 융합된 streptavidin moiety에 의해 monomer subunit가 비공유결합으로 tetramerization됨을 제시해준다. 또한 본 융합단백질은 ELISA와 Westernblot analysis에서 보여진 것처럼 parental streptavidin과 유사한 biotin결합력과 green fluorescent protein항원 결합력을 모두 나타내었다. 결론적으로 streptavidin에 융합된 8B9 VHH 항체형태의 융합단백질은 대장균에서 수용성 tetramer로 성공적으로 발현 및 정제되었으며 biotin과 green fluorescent protein 항원에 동시에 결합함으로써 tetrameric and bifunctional VHH 항체제조의 가능성을 제시해주었다.

• 자원환경지질
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• 제55권1호
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• pp.63-76
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• 2022

이산화탄소 지중저장 가능성 평가를 위해 시추한 결과, 장기분지 성동리층에 97~118m 두께의 데사이트질 응회암이 확인되었으며 내부구조와 입자조성에 따라 4개의 퇴적단위(퇴적단위 1~4)로 구분되었다. 퇴적단위 1은 육성환경, 퇴적단위 3, 4는 수중 환경에서 쌓인 화쇄류암(ignimbrite)이며, 퇴적단위 2는 화산휴지기에 쌓인 망상하천 퇴적암으로 데사이트질 응회암이 관찰되지 않는다. 박편분석결과, 모데나이트, 클리놉틸로라이트는 주로 유리를 교대하거나 기공을 충전하며 매몰-속성과정 동안 화산유리의 교대작용 및 용해-침전작용으로 생성된 것으로 보인다. 클리놉틸로라이트는 퇴적단위 3에서 유리를 교대하는 경우가 기공을 충전한 경우보다 Si/Al비와 Na함량이 높으며 퇴적단위 4에서는 산출상태 및 지역에 상관없이 조성이 동일해 지는데, 이는 화산유리의 교대 및 용탈작용이 진행될수록 공극수의 Si/Al비와 pH가 증가했으며, 퇴적단위 3의 클리놉틸로라이트가 생성될 당시 동-서쪽의 공극수 조성이 달랐음을 지시한다. 또한, 모데나이트의 성장이 끝난 후 클리놉틸로라이트가 생성되었으며, 두 불석광물은 각 퇴적단위별로 순차적으로 생성된 것으로 해석된다. 종합해보면, 퇴적단위 1이 쌓인 후, 화산휴지기 동안 서고동저의 분지지형으로 인해 서쪽은 pH가 더 높았으나 스멕타이트의 성장으로 공극이 폐쇄되어 더 이상 불석광물 이 성장하지 못한 반면, 동쪽은 망상하천이 발달한 개방계 상태에서 지하수의 영향을 받아 클리놉틸로라이트가 성장할 수 있었던 것으로 보인다. 이후 분지의 침강으로 퇴적단위 3, 4는 수중환경으로 변화였고 공극수는 점점 알칼리함량이 증가하여 결국 동-서쪽의 공극수 조성이 동일해 진 것으로 해석되었다. 이와 같이 유사한 조성의 응회암에서도 분지의 수계와 퇴적환경에 따라 다양한 불석광물이 공간적으로 다르게 분포할 수 있음을 보여준다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권9호
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• pp.1237-1247
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• 2012

The objective of this study was to evaluate the predictions of dry matter intake (DMI) and average daily gain (ADG) of Vietnamese Yellow (Vang) purebred and crossbred (Vang with Red Sindhi or Brahman) bulls fed under Vietnamese conditions using two levels of solution (1 and 2) of the large ruminant nutrition system (LRNS) model. Animal information and feed chemical characterization were obtained from five studies. The initial mean body weight (BW) of the animals was 186, with standard deviation ${\pm}33.2$ kg. Animals were fed ad libitum commonly available feedstuffs, including cassava powder, corn grain, Napier grass, rice straw and bran, and minerals and vitamins, for 50 to 80 d. Adequacy of the predictions was assessed with the Model Evaluation System using the root of mean square error of prediction (RMSEP), accuracy (Cb), coefficient of determination ($r^2$), and mean bias (MB). When all treatment means were used, both levels of solution predicted DMI similarly with low precision ($r^2$ of 0.389 and 0.45 for level 1 and 2, respectively) and medium accuracy (Cb of 0.827 and 0.859, respectively). The LRNS clearly over-predicted the intake of one study. When this study was removed from the comparison, the precision and accuracy considerably increased for the level 1 solution. Metabolisable protein was limiting ADG for more than 68% of the treatment averages. Both levels differed regarding precision and accuracy. While level 1 solution had the least MB compared with level 2 (0.058 and 0.159 kg/d, respectively), the precision was greater for level 2 than level 1 (0.89 and 0.70, respectively). The accuracy (Cb) was similar between level 1 and level 2 (p = 0.8997; 0.977 and 0.871, respectively). The RMSEP indicated that both levels were on average under-or over-predicted by about 190 g/d, suggesting that even though the accuracy (Cb) was greater for level 1 compared to level 2, both levels are likely to wrongly predict ADG by the same amount. Our analyses indicated that the level 1 solution can predict DMI reasonably well for this type of animal, but it was not entirely clear if animals consumed at their voluntary intake and/or if the roughness of the diet decreased DMI. A deficit of ruminally-undegradable protein and/or a lack of microbial protein may have limited the performance of these animals. Based on these evaluations, the LRNS level 1 solution may be an alternative to predict animal performance when, under specific circumstances, the fractional degradation rates of the carbohydrate and protein fractions are not known.

• 한국환경보건학회지
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• 제28권1호
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• pp.51-65
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• 2002

This study was carried out to apply some basic physical and chemical treatment options including Fenton's oxidation, and to evaluate the performances and the characteristics of organic and nitrogen removal using lab-scale biological treatment system such as complete-mixing activated sludge and sequencing batch reactor(SBR) processes for the treatment of leachate from a municipal waste landfill in Gyeongnam province. The results were as follows: Chemical coagulation experiments using aluminium sulfate, ferrous sulfate and ferric chloride resulted in leachate CO $D_{Cr}$ removal of 32%, 23% and 21 % with optimum reaction dose ranges of 10,000～15,000 mg/$\ell$, 1,000 mg/$\ell$ and 500～2,000 mg/$\ell$, respectively. Fenton's oxidation required the optimum conditions including pH 3.5, 6 hours of reaction time, and hydrogen peroxide and ferrous sulfate concentrations of 2,000 ～ 3,000 mg/$\ell$ each with 1:1 weight ratio to remove more than 50% of COD in the leachate containing CO $D_{Cr}$ between 2,000 ～ 3,000 mg/$\ell$. Air-stripping achieved to remove more than 97% of N $H_3$-N in the leachate in spite of requiring high cost of chemicals and extensive stripping time, and, however, zeolite treatment removing 94% of N $H_3$-N showed high selectivity to N $H^{+}$ ion and much faster removal rate than air-stripping. The result from lab-scale experiment using a complete-mixing activated sludge process showed that biological treatability tended to increase more or less as HRT increased or F/M ratio decreased, and, however, COD removal efficiency was very poor by showing only 36% at HRT of 29 days. While COD removal was achieved more during Fenton's oxidation as compared to alum treatment for the landfill leachate, the ratio of BOD/COD after Fenton's oxidation considerably increased, and the consecutive activated sludge process significantly reduced organic strength to remove 50% of CO $D_{Cr}$ and 95% of BO $D_{5}$ . The SBR process was generally more capable of removing organics and nitrogen in the leachate than complete-mixing activated sludge process to achieve 74% removal of influent CO $D_{Cr}$ , 98% of BO $D_{5}$ and especially 99% of N $H_3$-N. However, organic removal rates of the SBR processes pre-treated with air-stripping and with zeolite were not much different with those without pre-treatment, and the SBR process treated with powdered activated carbon showed a little higher rate of CO $D_{Cr}$ removal than the process without any treatment. In conclusion, the biological treatment process using SBR proved to be the most applicable for the treatment of organic contents and nitrogen simultaneously and effectively in the landfill leachate.e.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.220-220
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• 2013

Surface-normal transmission electro-absorption modulator (EAM) are attractive for high-definition (HD) three-dimensional (3D) imaging application due to its features such as small system volume and simple epitaxial structure [1,2]. However, EAM in order to be used for HD 3D imaging system requires uniform modulation performance over large area. To achieve highly uniform modulation performance of EAM at the operating wavelength of 850 nm, it is extremely important to remove the GaAs substrate over large area since GaAs material has high absorption coefficient below 870 nm which corresponds to band-edge energy of GaAs (1.424 eV). In this study, we propose and experimentally demonstrate a transmission EAM in which highly selective backside etching methods which include lapping, dry etching and wet etching is carried out to remove the GaAs substrate for achieving highly uniform modulation performance. First, lapping process on GaAs substrate was carried out for different lapping speeds (5 rpm, 7 rpm, 10 rpm) and the thickness was measured over different areas of surface. For a lapping speed of 5 rpm, a highly uniform surface over a large area ($2{\times}1\;mm^2$) was obtained. Second, optimization of inductive coupled plasma-reactive ion etching (ICP-RIE) was carried out to achieve anisotropy and high etch rate. The dry etching carried out using a gas mixture of SiCl4 and Ar, each having a flow rate of 10 sccm and 40 sccm, respectively with an RF power of 50 W, ICP power of 400 W and chamber pressure of 2 mTorr was the optimum etching condition. Last, the rest of GaAs substrate was successfully removed by highly selective backside wet etching with pH adjusted solution of citric acid and hydrogen peroxide. Citric acid/hydrogen peroxide etching solution having a volume ratio of 5:1 was the best etching condition which provides not only high selectivity of 235:1 between GaAs and AlAs but also good etching profile [3]. The fabricated transmission EAM array have an amplitude modulation of more than 50% at the bias voltage of -9 V and maintains high uniformity of >90% over large area ($2{\times}1\;mm^2$). These results show that the fabricated transmission EAM with substrate removed is an excellent candidate to be used as an optical shutter for HD 3D imaging application.

• 생태와환경
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• 제43권1호
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• pp.82-90
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• 2010

본 연구는 섬진강 중 상류 수계의 꺽지자원에 대해서 자원평가의 기초로 사용되는 자원생태학적 특성치들을 분석하고자 하였다. 연령사정을 위한 연령형질로서 이석(otolith)을 사용하였으며, 연령사정 결과, 최고연령은 5세로 나타났다. 체장(BL)과 체중(BW)의 관계식은 $BW=0.0195BL6{3.08}$($R^2=0.966$) 이었으며, 윤문이 형성되었을 때의 체장을 역계산하기 위한 체장(BL)과 이석경(R)과의 관계식은 BL=3.882R+1.66($R^2=0.944$)로 나타났다. 비선형회귀방법을 이용한 von Bertalanffy 성장모델의 매개변수는 이론적 최대체장($L_{\infty}$)이 19.68 cm, 이론적 최대체중($L_{\infty}$)이 188.64 g, 성장계수(K)가 0.17, 체장이 0 일 때의 연령이 -1.46세 등으로 각각 추정되었다. 이를 통해 추정된 성장식은 Lt=19.68(1-$e^{-0.17(t+1.46)}$)($R^2=0.997$)로 나타났다. 생존율을 추정하는 6가지 방법 중 평방오차합(Sum of squared error: SSQ) 이 가장 작은 어획물곡선법을 이용하여 생존율을 추정하였으며, 추정된 생존율(S)는 $0.666\;year^{-1}$으로 확인되었다. 순간자연사망계수(M)와 순간전사망계수(Z)는 $0.346\;year^{-1}$과 $0.407\;year^{-1}$로 각각 추정 되었으며, 이를 통해 확인된 순간어획사망계수(F)는 $0.061\;year^{-1}$로 확인되었다.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.448-468
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• 1996

To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

• 한국산림과학회지
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• 제83권3호
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• pp.286-298
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• 1994

본(本) 연구(硏究)는 덕유산(德裕山) 국립공원(國立公園)에서 가장 많이 이용되고 있는 매표소 백련사 향적봉-동엽령-칠연폭포 코스를 조사 대상지로 선정하여 조사 대상지를 매표소-백련사, 백련사-향적봉, 향적봉-동엽령, 동엽령-칠연폭포 등 4개 구간으로 구분하고 각 구간에서 등산로(登山路)의 훼손실태(毁損實態)와 향적봉(香績峯) 정상일대의 자연환경(自然環境) 훼손실태(毁損實態) 등을 조사(調査) 분석(分析)하였는 바, 그 결과(結果)를 요약하면 다음과 같다. 1. 매표소-백련사, 향적봉-동엽령, 동엽령-칠연폭포 구간은 경사가 완만한 편이나, 백련사-향적봉 구간은 $30^{\circ}$이상의 험준한 지형(地形)을 이루고 있다. 이용량(利用量)에 의한 이용강도(利用强度)는 매표소-백련사 구간은 "매우 강(强)하다", 백련사-향적봉 구간은 "강(强)하다", 향적봉-동엽령 구간은 "약(弱)하다"로 구분할 수 있었고 동엽령-칠연폭포 구간은 자연휴식년제(自然休息年制) 실시로 "이용하지 않는" 상태이다. 2. 등산로폭(登山路幅)은 전체 평균 4.3m이고 이용강도(利用强度)가 강(强)할수록 넓게 나타났으며, 백련사-향적봉 동엽령 구간에서는 배수불량지(排水不良地), 급경사지(急傾斜地) 등에서 등산로가 분기(分岐)되는 현상이 나타났다. 3. 등산로(登山路)의 평균 최대 침식(侵蝕) 깊이와 횡단면적(橫斷面積)은 백련사-향적봉 구간에서 각각 103.8cm, $42,972.9cm^2$로 최대치를 나타내었고, 등산로의 측면(測面) 붕괴(崩壞)도 가장 많아 이 구간의 침식(侵蝕)이 가장 심(甚)한 상태였다. 4. 백련사-향적봉 구간의 등산로는 바위와 라출근(裸出根)이 60%이상을 차지하고 있어서 노면(路面)의 상태가 대단히 불량(不良)하였다. 5. 토양(土壤)의 이화학적(理化學的) 성질(性質) 중 토양경도(土壤硬度), pH, 치환성염기(置換性鹽基)이온 (Na, K, Ca, Mg) 등은 등산로(登山路) 토양(土壤)에서 높았고 토양함유량(土壤含有率), 유기물함량(有機物含量), 전질소(全窒素), 유효인산(有效燐酸), C.E.C 등은 산림지(森林地) 토양(土壤)에서 높게 나타나 탐방객의 답압(踏壓)에 의한 영향이 뚜렷하였다. 또한 이용강도에 따른 변화가 뚜렷한 것은 토양경도는, 함수율, 유기물과 전질소 함량으로서 이중 토양경도는 이용강도가 증가할수록 높았고, 나머지 성질들은 이용강도가 증가할수록 작은 값을 나타내었다. 6. 정상(頂上)인 향적봉(香績峯) 일대에는 정상부, 헬기장, 야영장을 포함하여 약 $2,000m^2$의 라지(裸地)가 형성(形成)되어 있으며 토양조건(土壤條件)이 매우 불량(不良)한 상태이다. 7. 등산로의 환경훼손(環境毁損)을 유발하는 주요인은 이용강도(利用强度)와 지형적 특성(특히 경사도(傾斜度))인 것으로 나타났다. 8. 동엽령-칠연폭포 구간에서 자연휴식년제(自然休息年制) 실시의 효과(效果)가 부분적으로 인정되었다.

• 대한치과보철학회지
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• 제51권4호
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• pp.315-322
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• 2013

연구 목적: 내측연결형 임플란트와 지대주의 연결체에 반복하중을 부여하였을 때 수직 침하를 평가하고자 하였다. 연구 재료 및 방법: 외측연결형 임플란트와 내측연결형 임플란트에 세 종류의 시멘트유지형 지대주를 각각 장착하였다. 즉, 외측연결형 지대주(Ext 그룹), 내측연결형 1-piece 지대주(Int-1 그룹), 내측연결형 2-piece 지대주(Int-2 그룹)를 사용하였으며, 각 그룹마다 7개의 시편을 준비하였다. 임플란트-지대주 연결체에 수직하중을 적용하기 위하여 임플란트 받침대에 고정한 후, 4 Hz의 빈도로 $150{\pm}10N$의 반복하중을 가하였다. 수직침하량은 0, 5, 10, 50, 100, 1,000, 5,000, 10,000회의 반복하중 후에 각각 측정하였다. 반복측정분산분석(RM-ANOVA)를 이용하여 반복하중의 영향을 분석하였으며, 패턴변화를 관찰하기 위하여 선형혼합모형(linear mixed model)을 사용하였다. 유의수준은5% 로 설정하였다. 결과: 10,000회 반복하중 후 수직침하량은, Ext 그룹에서 $0.714{\pm}0.488{\mu}m$, Int-1그룹에서 $5.286{\pm}1.604{\mu}m$, Int-2 그룹에서 $11.429{\pm}1.902{\mu}m$를 나타내었다. 패턴분석에서는, Int-1 그룹 및 Int-2 그룹에서 지속적인 수직침하가 관찰되었으며, Ext그룹에서는 수직침하현상이 관찰되지 않았다. 결론:10,000회 반복하중 후의 선형혼합모형을 통한 분석에서, Ext그룹은 수직침하현상을 보이지 않았으나, Int-1 및 Int-2 그룹은 지속적인 수직침하현상을 나타내었다. 또한, Int-2그룹에서 Int-1그룹보다 더 많은 수직침하량이 관찰되었다.

• 대한미생물학회지
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• 제21권1호
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.