• Parasites, Hosts and Diseases
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• 제37권3호
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• pp.163-169
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• 1999

House dust mite allergens have been well established as sensitizing agents that are important in the induction of allergic diseases. In order to analyze epitopes of the allergen and to develop a quantitative method of the allergen exposure, monoclonal antibodies against a recombinant Der p 2 (rDer p 2), one of the major allergens of Dermatophogoides pteronyssinus, were produce. Four monoclonal antibodies produced wee species-specific and did not cross-react to the D. farinae crude extract. Two of the monoclonal antibodies were found to be IgG1 and the others were IgM. For the analysis of epitopes, a Der p 2 cDNA encoding 126 amino acids (aa) was dissected into three fragments with several overlapping peptides, A (aa residues 1-49), B (44-93), and C fragment (84-126). Three monoclonal antibodies showed reactivities to the recombinant B fragment and to the full-length rDer p 2, but one monoclonal antibody reacted only with the full-length rDer p 2. Two-site capture ELISA was developed using two different monoclonal antibodies for quantitating Der p2 in house dust. The sensitivity limit was 4ng/ml with rDer p2 and $8{\;}\mu\textrm{g}/ml$ with the d. pteronyssinus crude extract. The result suggested that the assay using monoclonal antibodies against rDer p2 could be useful for the environmental studies and for the standardization of mite allergen extracts.

• 대한핵의학회지
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• 제24권1호
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• pp.29-36
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• 1990

Most of the diagnostic methods currently used for the detection of neoplastic masses provide indirect evidence. To obtain greater specificity in the interpretation of neoplasias by in vivo methods, the immunological approach appears to be most promising. Two problems that interfered with progress in this field were the lack of tumor specific antigen and the lack of well-defined and reproducible antibodies. To improve the sensitivity and specificity of radioimmunoscintigraphy as a technique for tumor localization, the use of monoclonal antibodies, fragments of antibodies and single photon emission computerized tomography (SPECT) are reasonable. The obvious advantages of monoclonal antibodies are their homogeneity, their specificity for the immunizing antigen and the reaction with a single determinant-thus no large immunecomplexes with antigen are formed. Monoclonal antibody technique has recently provided an opportunity to reevaluate the role of nuclear medicine for the diagnosis of malignant diseases by using the immunological approach. Out first results by means of radioimmunoscintigraphy of CEA and CA 19-9 producing tumors using a cocktail of fragments F $(ab')_2$, of mocolonal antibodies to CA 19-9 and CEA labeled with $^{131}I$ (IMACIS-1) are reported. The aims of this investigation was to evaluate the role of immunoscintigraphy in patients with colorectal and other cancers for diagnosis of local recurrences and metastasis. This report contains results of the first 8 colorectal and pancreas cancer patients with the elevation of the level of serum CEA and/or CA 19-9. IMACIS-1 was injected intravenously during 30 minutes in 100 ml saline solution after skin test. Planar scintigrams were recorded 3, 5 and 7 days after the injection of the IMACIS-1. Anterior, lateral and posterior views of the liver as well as anterior and posterior views of the pelvis were obtained in each patients as an $^{131}I-antibody$ image. We were able to localize exactly the malignant process with the double-nuclide double-compound $^{99m}Tc\;^{131}I$ (Tc+l) scintigrams. In Tc & I double-nuclide scintigraphy, computer subtraction display provided more clear localization of the tumor. We compared the results of radioimmunoscintigraphy with CT, ultrasonograms, conventional scintigrams. The results were as follows: 1) The sensitivity and specificity of radioimmunoscintigraphy using the fragments $F(ab')_2$ of the cocktails of CEA and CA 19-9 monoclonal antibodies were 80% and 100% respectively. 2) Tumor detection rate was not proportionated to the level of serum tumor markets. 3) Second tracer technique was essential for tumor localization as an anatomic landmark using double-nuclide scintigraphy. 4) A slow infusion of the antibodies was necessary to prevent the formation of large immune complexes. 5) Tumor/non-tumor radioactivity was most elevated at 7 days delayed imaging. 6) Using planar scintigraphic technique of $^{131}I$ labeled monoclonal antibodies are possible for imaging most of the tumors.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.102-102
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• 2001

Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

• Journal of Animal Science and Technology
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• 제51권5호
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• pp.407-412
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• 2009

본 연구는 송아지 설사병의 주요원인체 중 소로타바이러스(bovine rotavirus; BRV)와 소코로나바이러스(bovine coronavirus; BCV)에 대한 난황항체를 생산하고 이의 면역 특이성을 확인하고자 실험을 실시하였으며, BRV 및 BCV를 2주 간격으로 5회 산란계에 근육주사를 실시하여 혈청과 난황내의 특이 항체 형성 유무를 확인하였다. 실험 6주차에 면역한 산란계로부터 획득한 혈청을 이용하여 교차반응 시험을 실시한 결과, BRV 및 BCV를 면역하여 얻은 혈청은 각각 BRV 및 BCV 항원과만 특이적인 결합반응을 보였다. 면역에 따른 혈청항체 및 난황항체의 수준은 실험 8~12주차에 최고도에 달했고, BRV에 대한 항체의 경우 혈청과 난황내에서 면역 후 12주째에 각각 약 104,000과 107,000의 역가 수준을 보였으며, BCV의 경우 8주째에 각각 약 145,000과 155,000의 수준을 보였다. BRV 및 BCV에 대한 중화능력 유무 확인을 위하여 분리된 난황항체를 이용하여 혈구응집억제반응 시험을 실시한 결과, BRV 및 BCV에 대한 혈구응집억제 희석비가 각각 5,120 및 1,260으로 면역하지 않은 대조군에 비하여 8배 이상 높은 중화력을 나타냈다. 이러한 결과를 종합하면, 산란계에 BRV 및 BCV를 면역하여 얻어진 난황항체는 BRV 및 BCV에 대한 면역 특이성을 가지고 중화할 수 있는 능력이 있으며, 이러한 난황항체는 BRV 및 BCV의 증식을 효과적으로 억제시킬 수 있어 임상에 적용할 경우 유용할 것으로 사료된다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 제34회 학술심포지움
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• pp.31-32
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• 2001

• 한국동물학회:학술대회논문집
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• 한국동물학회 1998년도 한국생물과학협회 학술발표대회
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• pp.244.1-244
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• 1998

No Abstract, See Full Text

• 대한수의사회지
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• 제17권3호
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• pp.47-60
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• 1981

Lymphocytes that secrete antibodies can be made immortal by fusing them with myeloma tumor cells (cell fusion) and cloning the hybrids (hybridomas). Each clone is a long-term source of substantial quantities of a single highly specific antibody (monoclona

• Pediatric Infection and Vaccine
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• 제12권2호
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• pp.108-113
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• 2005

목 적 : 최근 HHV-8는 면역력이 정상인 소아에서 원발성 감염에 의해 발진이 동반된 발열성 질환을 유발하는 것으로 알려지고 있다. 본 연구는 국내 정상 소아의 항 HHV-8 항체 양성률을 알아보기 위해 시행되었다. 방 법 : 항 HHV-8 항체의 양성률을 파악하기 위하여 인제의대 상계백병원을 방문한 환자 중에서, 최근 3주간 발열 증상이 없었던 건강한 소아 112명의 혈청을 대상으로 하였다. Lytic 바이러스 항원에 대한 IgG 항체 검사는 간접면역 형광검사법, HHV-8의 ORF에 대한 특이 항체 검사는 peptide mix ELISA 검사법을 사용하였다. 결 과 : 대상 환아는 총 112명이었으며 남아 64명, 여아 48명이었다. 1세 이하의 연령군은 17명이었으며, 2~5세 연령군 24명, 6~9세 연령군 24명, 10~15세 연령군은 47명이었다. HHV-8에 대한 혈청 항체 양성률은 간접 면역 형광검사법으로 3.5%(4/112), ELSIA로는 1.8%(2/112)였다. 결 론 : 국내 소아의 HHV-8 항체의 양성률은 비교적 낮았지만 앞으로 발진이 동반된 발열성 질환에서 HHV-8의 역할에 대한 추가적인 연구가 필요할 것으로 보인다.

• 한국식품위생안전성학회지
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• 제31권6호
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• pp.445-450
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• 2016

본 연구에서는 돈지육 및 돈육 조직 내에 열안정성 수용성 단백질의 존재 여부를 확인하고 항체 생산에 있어 항원으로의 사용 가능 여부를 확인하고자 하였다. 이를 위해 돈지육 및 돈육을 생(raw) 시료와 조리된(cooked) 시료로 구분하여 비열처리 및 열처리법으로 단백질을 추출한 후 단백질 존재여부를 단백질 정량과 SDS-PAGE로 확인하였다. 그 결과 돈지육과 돈육 모두 생 시료를 비열처리법으로 추출한 시료의 경우 25~100 kDa 사이의 다양한 단백질이 확인된 반면 시료를 가열하거나 추출 시 열처리를 한 경우 돈지육에는 100 kDa 이상의 단백질과 30 kDa 및 15 kDa 이하의 일부 단백질이, 돈육에는 100 kDa 이상과 30 kDa 이하의 단백질이 확인되어 돈지육과 돈육에 열안정성 수용성 단백질이 존재하는 것으로 확인되었다. 이들 열안정성 수용성 단백질을 마우스에 면역 후 항혈청 역가를 측정한 결과 면역한 모든 마우스에서 높은 역가를 나타내었고, 생산된 혈청은 돈지육과 돈육에 각각 특이적인 반응성을 보인 반면 다른 축육과 지방육에 대해서는 반응성이 상대적으로 낮았다. 이러한 연구결과를 볼 때 돈지육 및 돈육에 존재하는 열안정성 수용성 단백질이 돈지육과 돈육에 특이적으로 반응하는 항체를 개발하는데 유용한 마커로서 활용이 가능하며, 열안정성 수용성 단백질에 대한 항체개발은 열처리된 축육 가공품 중 돈지육 및 돈육의 분석에도 매우 유용하게 활용할 수 있을 것으로 판단된다.

• Food Science and Biotechnology
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• 제16권4호
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• pp.515-519
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• 2007

Rapid immunochromatographic assay (ICA) kits were developed using flagella-specific monoclonal antibodies (MAbs) and rabbit polyclonal antibodies for screening Listeria spp. in food. The establishment of different formats, MAb 2B1 as capture antibody and MAb 7A3 or rabbit polyclonal antibodies as detector antibody, was compared. The 2 formats of the ICA kit were shown to have specific reactions with Listeria and no cross-reactivity with any of the non-Listeria including Escherichia coli O157:H7 and Salmonella enteritidis. The detection limits of the ICA kit using the combination of gold-labeled MAb 7A3 and MAb 2B1 showed $1{\times}10^5$ and $1{\times}10^6\;CFU/0.1\;mL$ at 22 and $30^{\circ}C$, respectively. The other format of the ICA kit using the combination of gold-labeled rabbit polyclonal antibodies and MAb 2B1 showed $1{\times}10^6\;CFU/0.1\;mL$ at $22^{\circ}C$ but weak signal at 30 culture. The format utilizing MAb was more sensitive than the one using polyclonal antibodies for capture antibody. Samples contaminated with L. monocytogenes 4b culture (9-10, 5-6, and 1-2 CFU/mL) on pork and pasteurized milk were confirmed as positive results. Current data suggests that this ICA kit is a rapid, simple and effective tool to screen for Listeria spp. in food.