• 대한미생물학회지
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• 제18권1호
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• pp.73-85
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• 1983

The advantages of the enzyme-linked immunosorbent assay(ELISA) are its sensitivity and its simplicity in detecting IgM and IgG antibodies. For applying the ELISA to the diagnosis of typhoid fever, first of all, experiments were performed to determine which concentration of killed whole cell antigens and lipopolysaccharide(LPS) antigens of S. typhi(0.901 w) were optimally coated to the wells of the polystyrene and polyvinylchloride microplate, using the hyperimmune sera from rabbits against S. typhi. By using both kinds of antigens of S. typhi adsorbed to the ELISA microplate, the changing patterns of IgG and IgM antibodies in the sera from rabbits responding to the killed whole cell antigens of S. typhi(0901 w) during the prolonged immunization were serially traced by the ELISA. At the same time, the level of antibodies against S. typhi in sera fron patients with typhoid fever and from normal healthy persons were measured by the ELISA employing the killed whole cell antigens and LPS antigens as the coating antigens. The results obtained were summerized as follow: 1. The optimal concentration of the killed whole cell antigens, which were more easily adsorbed to the polystyrene plate than the polyvinylchloride plate, was $10^8cells/ml$ of carbonate buffer(pH. 9.6) on the wells of the polystyrene plate when treated at $37^{\circ}C$ for 4 hours. On the other hand, the optimal concentration of lipopolysaccharide antigens, which were adsorbed only to the polyvinylchloride plate, was $100{\mu}g/ml$ of carbonate buffer(pH. 9.6) on the wells of the polyvinylchloride plate when treated at $37^{\circ}C$ for 4 hours. 2. IgM antibody response were dominating in rabbits responding to the killed whole cell antigens of S. typhi(0.901 w), and were more specific to the LPS antigens than to the killed whole cell antigens in the ELISA. Good correlations were made between the IgM titers by the ELISA and the aggglutinating titers of sera from the immunized rabbits. 3. Both IgG and IgM agglutination titers by the ELISA in sera from most of patients with typhoid fever were above 1:320 but those in sera from most of normal, healthy persons were below 1:80. 4. There were close correlations between the antibody titers by the ELISA and the agglutinating titers to the killed whole cell antigens in the tested human sera, IgM titers being more correlated with the agglutinating titers than IgG titers. But a little correlations were made between the antibody titers by the ELISA and the agglutinating titers to the LPS antigens. 5. IgM titers in the tested human sera were similar to IgG titers detected by the ELISA employing the killd whole cells antigens and the LPS antigens. 6. Good correlations were made between the antibody titers demonstrated by the ELISA performed on the killed whole cell antigens and the LPS antigens as the different, coating antigens on the ELISA microplates.

• Parasites, Hosts and Diseases
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• 제33권3호
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• pp.225-230
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• 1995

저자등은 1994년 5월 경남 마산시에 거주하는 41세 남자 환자에게서 자연배출된 의엽 조충류의 충체 일부를 고래복식문조충(Diplogonoporus balaenopterae)으로 동정하였고 우리 나라의 인체 감염 제1례로 보고하는 바이다. 환자는 평소 각종 해산어류의 생식을 즐기고 담수어로는 잉어만을 생식한 경험이 있다. 검사실 소견상 경미한 호산구증다증(6%)과 혈청 IgE의 심한 증가(10,182 IU/ml). 초음파상에서 알콜성 지방간 외에는 특이한 소견을 발견할 수 없었다. Niclosamide 2 g을 경구투여하여 구충하였으나 두절을 발견하지 못하였다.

• 미생물학회지
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• 제51권2호
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• pp.115-125
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• 2015

단일클론항체 AP64 IgM은 인간의 급성 비임파성 골수암(ANLL) 세포주 HL60에 결합하며 쥐의 ANLL 세포에도 교차결합(cross-react)한다. 또한 complement에 의해 매개되어지면 골수암 억제효과를 나타낸다. 본 연구에서는 RT-PCR에 의해 AP64 IgM을 분비하는 하이브리도마의 $V_H$ 및 $V_L$ cDNA로부터 유래된 재조합 single-chain variable domain fragment (ScFv)를 제조하였다. $V_H$ 및 $V_L$은 15개 아미노산으로 구성된 linker $(G_4S)_3$으로 연결되었다. 재조합 ScFv는 Escherichia coli BMH 71-18에서 single polypeptide chain으로 발현되었다. Periplasmic extract를 $Ni^+$-NTA-agarose affinity column에 가하여 발현된 재조합 ScFv를 정제하였으며 westernblot으로 정제된 단백질을 탐지하였다. 정제된 재조합 ScFv는 AP64 IgM 모항체가 탐지하는 항원과 같은 HL60 세포의 표면항원(약 30 kDa)을 인지하였다. 그러나 HL60의 표면항원에 대한 ScFv의 결합력은 AP64 IgM 모항체보다 낮아서 추후 이에 대한 개선이 필요하다. 종합하여 볼 때 HL60 세포주에 특이적인 재조합 ScFv는 진단 또는 치료목적으로 유용한 생물학적 제재가 될 수 있을 것이다.

• Parasites, Hosts and Diseases
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• 제26권1호
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• pp.1-8
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• 1988

프라지관텔이 사용되어 많은 간흡충 감염자가 치료되고 있는 현재의 상환에 비추어, 간흡충중의 진단에서 피내반응검사와 혈청검사의 효용성과 치료 후의 혈청 항체가의 변동 양상을 관찰하고자 이 연구를 실시하였다. 그 결과를 요약하면 다음과 같다. 1. 피내 반응검사를 실시한 135명 중에서 간흡충의 VBS항원에 양성 반응자가 35명 (25.9%)이었고 이 중에서 간흡충란 양성자는 12명 (34.3%)이 었다. 2. 전체 간흡충란 양성자 129명에서 간흡충 성충의 조항원을 이용한 효소면역 법(ELISA)의 흡광도가 0.063~1.216($0.325{\pm}0.202$)이 고 25명 의 충란음성 자에 서 는 0.078~0.670($0.255{\pm}0.133$)이었다. 3. 총 93례에서 실시한 EPG와 휴광도의 관계를 보면 68명의 EPG 900이하 감염례에서 0,063~0.761($0.258{\pm}0.142$)이고, EPG 1,000~1,900의 6례에서 0,231~0.773($0.463{\pm}.191$0), EPG 2,0000~3,900의 6례에서 0.361~0.640($0.497{\pm}0,103$), EPG 4,000~9,900의 7례에서 0.196~0.874($0.517{\pm}0.261$), 6명의 중감염자(EPG 10,000 이상)에서 0.359~1.216($0.715{\pm}0.342$)이었다. 4. 치료 전후에 관찰한 혈청내 특이 IgG항체가는 각 군에 따라서 다음과 같다. 제 1군의 치료전 흡광도는 0.075~1.216($0.347{\pm}0.258$)이고 치료 12~14일 후에는 0.065~l.181($0.417{\pm}0.272$)이었고, 제 II군의 홉광도는 0.IG2~10.796($0.320{\pm}0.287$)이고 치료 4주 후에 0.107~0.544($0.206{\pm}0.115$)이었다. 제III군에서는 흡광도가 치료 전에 0.102~0.470($0.244{\pm}0.117$)이며 치료 8~9주 후에 0.101~0.500($0.256{\pm}0.134$)이었다. 제 IV군의 경우 흡광도가 0.063~0.735($0.320{\pm}0.189$)에서 치료 후 7개월에 0.090~0.404($0.201{\pm}0.088$)로 감소하고, 제 V군에서 흡광도 0.063~0.773($0.352{\pm}0.227$)이 치료 13개월 후에 0.076~0.386($0.216{\pm}0.096$)으로 감소하였다. 치료 전후의 흡광도는 7개월 및 13개월 후의 변화만이 통계적으로 유의하였다. 이상의 결과를 미루어보면 간흡충증의 진단에서 효소면역 법을 이용한 혈청 내 특이 IgG항체 검사는 EPG 1,000 이상의 중등도 이상 감염자에서만 우수한 진단적 가치를 가진다고 판단된다. 또한 프라지관델로 치료하여 충란음전이 된 상태에서 혈청내 ITG항체는 투약 후 9주와 7개월 사이에 유의하게 감소하였다.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• Parasites, Hosts and Diseases
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• 제45권4호
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• pp.287-293
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• 2007

The identification and characterization of antigens that elicit human T cell responses is an important step toward understanding of Leishmania major infection and ultimately in the development of a vaccine. Micropreparative SDS-PAGE followed by electro transfer to a PVDF membrane and elution of proteins from the PVDF, was used to separate 2 novel proteins from L. major promastigotes, which can induce antibodies of the IgG2a isotype in mice and also are recognized by antisera of recovered human cutaneous leishmaniasis subjects. Fractionation of the crude extract of L. major revealed that all detectable proteins of interest were present within the soluble Leishmania antigens (SLA). Quantitation of these proteins showed that their expression in promastigotes is relatively very low. Considering the molecular weight, immunoreactivity, chromatographic and electrophoretic behavior in reducing and non-reducing conditions, these proteins are probably 2 isoforms of a single protein. A digest of these proteins was resolved on Tricine-SDS-PAGE and immunoreactive fragments were identified by human sera. Two immunoreactive fragments (36.4 and 34.8 kDa) were only generated by endoproteinase Glu-C treatment. These immunoreactive fragments or their parent molecules may be ideal candidates for incorporation in a cocktail vaccine against cutaneous leishmaniasis.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.84-84
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• 2013

FcBP consisting of 13 amino acids specifically binds to Immunoglobulin G Fc domain. Initially, we utilized this peptide for preparation of antibody chip as a PEG composite for enhanced solubility. After then, the peptide conjugate was immobilized on agarose resin, resulting in highly efficient affinity column for antibody purification. The efficiency was comparable to commercial Protein A column. Recently, this peptide was conjugated with cell penetratingpeptide (CPP) on a backbone of GFP, affording antibody transducer, which carries antibody into live cells by simple mixing of antibody and the transducer in cell culture media. Antibody transduction into cells was monitored by live cell imaging. More recently, the FcBP was fused to ferritin cage, which consists of 24 ferritin protein molecules. The FcBP-ferritin cage showed greatly increased binding affinity to human IgG. Its binding was analyzed by QCM and SPR analysis. Finally, it was selectively delivered by Herceptin to SKBR3, a breast cancer cell, over MCF10A, non-tumorigenic cells.

• BMB Reports
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• 제43권2호
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• pp.146-149
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• 2010

Exendin-4 (Ex-4), a peptide secreted from the salivary glands of the Gila monster lizard, can increase pancreatic $\beta$-cell growth and insulin secretion by activating glucagon-like peptide-1 receptor. In this study, we expressed a fusion protein consisting of exendin-4 and the human immunoglobulin heavy chain (Ex-4/IgG-Fc) in E. coli and explored its potential therapeutic use for the treatment of insulin-resistant type 2 diabetes. Here, we show that the Ex-4/IgG-Fc fusion protein induces expression of insulin receptor substrate-2 in rat insulinoma INS-1 cells. Our findings therefore suggest that Ex-4/IgG-Fc overexpressed in E. coli could be used as a potential, long-acting glucagon-like peptide-1 mimetic.

• 대한한방소아과학회지
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• 제22권3호
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• pp.105-137
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• 2008

Objectives : The purpose of this study is to investigate the effect of Gupoongjeseuptang (GPJST) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of GPJST on atopic dermatifis, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells (PBMCs), splenocytes, draining lymph node (DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results : In vivo, clinical skin severity score were significantly lower in GPJST group than control group. IgE, IL-6, $TNF-{\alpha}$, IgG1, IgM, IgG2a and IgG2b levels in serum decreased remarkably in GPJST group than control group. Also, total absolute number of $CD3^+CD69^+$, and $CCR3^+$ cells recovered as normal in PBMCs and $CD3^+$, $CD3^+CD69^+$ decreased significantly compared with control group in isolated DLN from NC/Nga mouse and total absolute number of $CD11b^+Gr-1^+$, $CCR3^+CD3^+$ in dorsal skin of NC/Nga mouse decreased by GPJST. We analyzed ear and neck-back skin after biopsy and dyeing by hematoxyline/eosin (H&E) and toluidine staining (mast cells marker) and obtained results that GPJST are very effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell (CD4, $CCR3^+$) infiltration). Conclusions : This study demonstrates immunological activity of GPJST on atopic dermatitis-like model mice.

• 한국식품과학회지
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• 제40권3호
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• pp.343-348
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• 2008

여러가지 방법으로 난백을 처리하고 난백 중 주요한 알레르겐인 OM의 알레르기성 변화를 계란 알레르기 환자의 IgE 항체를 이용한 간접경합 ELISA로 조사하였다. 12종의 효소를 이용한 가수분해, 방사선조사, succinic anhydride 처리는 OM의 알레르기성을 효과적으로 감소시키지 못 하였다. OM의 당쇄를 제거하는 TFMS의 처리는 OM의 알레르기성을 1/20 정도로 감소시켰다. $100^{\circ}C$에서 10분간의 열처리는 알레르기성을 감소시키지 못하였으나 $121^{\circ}C$(10분)의 처리는 난백 중 OM의 알레르기성을 1/100정도로 감소시켰다. NaOH 단독 처리는 3%(w/v) 이상에서 알레르기성이 대부분 사라졌고, NaOH(0.3%(w/v) 이상)와 열($70^{\circ}C$,15 min)의 복합처리는 알레르기성을 1/10,000 이하로 낮춤으로써 가장 효과적인 감소를 보였다. 본 연구에서 환자의 IgE 항체를 이용한 ELISA 분석결과와 동일 시료에 대한 토끼 IgG 항체의 ELISA 결과는 대체적으로 비슷한 결과를 보였지만, TFMS 처리물에 대한 반응성은 후자에서 훨씬 많이 감소되어(1/10,000) 대조적이었다. 이는 식품의 항원성과 알레르기성 간에는 약간의 차이가 있음을 시사하였다.