• 제목/요약/키워드: Enzyme immunoassay

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모기유충에 살충력이 있는 Bacillus thuringiensis subsp. darmstadiensis 73E10-2 내독소의 면역학적 성질 (Immunological Characteristics of Mosquitocidal Delta-endotoxin from Bacillus thuringiensis Subsp. darmstadiensis 73E10-2)

  • 정태영;김광현
    • 한국미생물·생명공학회지
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    • 제18권3호
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    • pp.301-304
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    • 1990
  • Bti와 Btd의 내독소에 대한 면역학적인 검토를 행한 결과 양균주의 내독소에는 동일한 기원의 단백질이 혼존하고 있음을 면역확산반응 및 면역전기영동법으로 확인되었다. 또한 동일한 기원의 단백질 함량은 Bti 내독소 보다 Btd 내독소에 많이 존재함을 효소항체법으로 확인하였다.

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질병진단을 위한 나노자임 연구의 최근 동향 (Recent Advances in Nanozyme Research for Disease Diagnostics)

  • 신호연;윤태영;김문일
    • KSBB Journal
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    • 제30권1호
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    • pp.1-10
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    • 2015
  • Nanomaterial-based artificial enzymes (Nanozymes) have attracted recent attention because of their unique advantageous characteristics such as excellent robustness and stability, low-cost production by facile scale-up, and longterm preservation capability that are critically required as an alternative to natural enzymes. These nanozymes exhibit natural enzyme-like activity, and they have been applied to diverse kinds of detection methods for disease-associated biomolecules such as DNAs, proteins, cells, and small molecules including glucose. To highlight the progress in the field of disease diagnostics using nanozyme, this review discusses many nanozyme-based detection methods categorized by the types of target biomolecules. Finally, we address the current challenges and perspectives for the widespread utilization of nanozyme-based disease diagnostics.

Implementing Expanded Rapid Human Immunodeficiency Virus Testing in Public Health Centers in Seoul, 2015

  • Kang, Cho Ryok;Bang, Ji Hwan;Cho, Sung-Il;Lee, Young Hwa;Oh, Myoung-don;Lee, Jong-Koo
    • Infection and chemotherapy
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    • 제50권4호
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    • pp.346-349
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    • 2018
  • In 2015, rapid human immunodeficiency virus (HIV) testing was implemented in all 25 public health centers in Seoul. During March and December 2015, 20,987 rapid HIV tests were performed, of which 116 (0.5%) were positive. Compared to those of the period before application of the rapid HIV test in place of conventional enzyme immunoassay method, the number of HIV tests performed and the number of positive results increased by sevenfold and twofold, respectively. In conclusion, expansion of the provision of rapid HIV tests in public health centers increased the number of voluntary HIV tests.

Detection of Serum IgE Specific to Mite Allergens by Immuno-PCR

  • Lee, Kyung-Woo;Hur, Byung-Ung;Chua, Kaw-Yan;Kuo, I-Chun;Song, Suk-Yoon;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • 제8권3호
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    • pp.82-89
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    • 2008
  • Background: Although a skin test is the primary option for detecting allergen-specific IgE in clinics, the serum IgE immunoassay is also important because it allows for the diagnosis of allergy without any accompanying adverse effect on the patient. However, the low detection limit of IgE levels by immunoassay may restrict the use of the method in some occasions, and improving its sensitivity would thus have a significant implication in allergy-immunology clinics. Methods: In this study, we attempted to detect specific serum IgE by using immuno-polymerase chain reaction (IPCR) which combines the antigen-antibody specificity of enzyme-linked immunosorbent assays (ELISAs) with the amplification power of PCR. Results: Our results demonstrated that Blo t5-specific serum IgE can be detected by IPCR with a 100-fold higher sensitivity than ELISA, and cross-reactivity of serum IgE to other mite allergens is able to be analyzed by using only $0.3{\mu}l$ of serum sample. Use of real-time IPCR seemed to permit more convenient determination of specific serum IgE as well. Conclusion: We believe that IPCR can serve as a valuable tool in determining specific serum IgE, especially when the amount of serum sample is limited.

면역 조직화학 반응이 통합된 바이오칩의 전기화학 및 광학적 분석 (Integration of immunohistochemical reactions into Electrochemical and Optical Analyses of Biochips)

  • 최형길;홍은경;이승원;윤현철
    • KSBB Journal
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    • 제20권2호
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    • pp.123-128
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    • 2005
  • 효율적인 바이오칩을 개발을 위해 칩 표면에 생체 물질들의 상호반응이 효과적으로 일어날 수 있는 센싱 표면의 조성과 항원-항체 반응과 같은 생체인식 반응을 정량적 신호로 전환하는 방법에 대해 연구하였다. 전기화학식 센서의 표면을 개선하기 위해 폴리아미도아민 덴드리머를 가교 물질로 도입하였다. 생체 분자들의 인식작용을 정량적인 신호로 전환하기 위해 전형적인 면역조직화학분석에서 사용된 반응들을 바이오센서에 적용한 방법론을 사용하였다 효소에 의해 촉매되는 신호화 방법은 면역반응들에 대하여 광학식 센서와 전기화학식 센서에서 공히 수행되었으며, 매우 정량적인 신호로 측정되었다. 측정된 신호들로부터 단백질 농도에 비례하는 검량곡선을 획득할 수 있었으며 다양한 면역 샘플에 대한 적용 가능성을 제시하였다.

Effects of Liriope Platyphylla on LPS-stimulated Expression of COX-2 and iNOS in Mouse BV2 Microglial Cells

  • Park, Sang-Heup;Kim, Ee-Hwa;Park, Se-Keun;Jang, Mi-Hyeon;Choi, Sun-Mi;Lee, Eun-Yong
    • Journal of Acupuncture Research
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    • 제22권2호
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    • pp.147-154
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    • 2005
  • Objective: In this study, the effects of Liriope Platyphylla against LPS-induced inflammation was investigated. Methods: Cell viability was determined using the MTT assay. To identify expressions of COX-2 and iNOS mRNA, RT-PCR was performed. Assessment of PGE2 synthesis was performed using the PGE2 immunoassay. Measurement of NO synthesis was performed using the NO detection. Result : The MTT assay revealed that Liriope Platyphylla exerted no significant cytotoxicity in the microglial BV2 cells. RT-PCR analysis revealed that the mRNA levels of COX-2 and iNOS were significantly decreased in the LPS- and 5 mg/ml Liriope Platyphylla treated group. From the PGE2 immunoassay and NO detection, PGE2 and NO synthesis was significantly suppressed in the LPS- and 5 mg/ml Liriope Platyphylla treated group. Conclusion : In these study, Liriope Platyphylla was shown to suppress PGE2 and NO production by inhibiting LPS-stimulated enhancement of COX-2 enzyme activity and iNOS expression. It is very possible that Liriope Platyphylla can offer a valuable mode of therapy for the treatment of brain inflammatory diseases.

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Production and Characterization of DDT Antibodies and Its Application to Enzyme Immunoassay: Relation of Response and Affinity to Coating Ligand

  • Hong, Ji-Youn;Kim, Jong-Hyun;Park, Song-Ja;Lho, Dong-Seok;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • 제24권11호
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    • pp.1605-1608
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    • 2003
  • To development an immunodetection method for DDT, 1,1,1-trichloro-2,2-bis(4-chorophenyl)ethane (p,p'-DDT) and its metabolites (p,p'-DDA, p,p'-DDE, p,p'-DDD), five derivatives of DDT haptens have been synthesised and characterized as coating ligands for antibody evaluation. The appropriate lengths of linkers were introduced to investigate a matching pair of coating ligand and antibody. Among these hapten derivatives, 2,2-bis(4-chlorophenyl)acetic acid (DDA), 5,5-bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for its use as an immunogen. The bovine serum albumin (BSA) conjugates of these derivatives were prepared as a coating ligand for monoclonal antibody screening. Fifteen monoclonal antibody clones were screened using these probes. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP), in addition to the above hapten derivatives, were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for their use as coating ligands to measure the titration level of the antibody and the displacement of free analytes. The indirect competitive ELISA results indicate that the titration level and free analyte displacement were greatly influenced by the DDT derivatives and carrier proteins used. Three matching pairs of monoclonal antibodies and coating ligands were selected for the DDT immunoassay: antibody clone 1A3 and coating ligand DDA-OVA, 1A1 and DDHHAP-BSA, and 1A4 and DDHP-OVA.

경쟁적 효소면역측정법을 이용한 환자 혈청 내 Thyroxine (T$_4$)의 검출 (Studies on the Rapid and Competitive Enzyme-linked Immunosorbent Assay for the Detection of Thyroxine (T$_4$) in Human Sera)

  • Sang-Wook Park;Jong-Bae Kim
    • 대한의생명과학회지
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    • 제5권1호
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    • pp.11-15
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    • 1999
  • 갑상선 질환의 진단에 필요한 갑상선 호르몬의 측정시 thyroxine (T$_4$)농도 측정방법이 가장 흔히 이용되고 있다. 과거에 많이 활용되어온 방사면역측정법 (RH)이 폐기물 처리 및 관리인원 등의 인적, 물적인 문제가 크게 대두되고 있는 점을 감안하면 효소결합면역측정법의 확립은 큰 의미가 있다고 하겠다. 본 연구에서는 thyroxine (T$_4$)에 대한 단클론 항체 (monoclonal antibody)를 이용하여 T$_4$에 horseradish-peroxidase를 화학적으로 결합시킨 microtiter plate에 HRP-conjugate T$_4$와 혈청내의 T$_4$간의 서로 경쟁적인 반응을 이용한 표준곡선을 얻었다. 그리고 이렇게 확립한 기법을 임상적인 갑상선질환 환자의 혈청을 기존의 chemiluminescence 방법을 이용하여 얻은 결과와 비교하여 본 실험에서 확립한 ELISA검사 수치의 유용성을 검토하였으며 이 방법의 임상이용 가능성을 검토하였다.

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${\gamma}$-Glutamyltransferase의 조직내 분포에 관한 연구 -단일클론항체의 효소면역측정법, 방사면역측정법, 면역조직화학검사, 자가방사기록검사 적용에 관하여 - (Distribution of Murine Tissue Specific ${\gamma}$-Glutamyltransferase: -Comparison of Six Monoclonal Antibody Applications in Enzyme Linked Immunosorbent Assay, Radioimmunoassay, Immunohistochemistry, and Autoradiography-)

  • 김명근;박윤규;류총근
    • 대한핵의학회지
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    • 제28권1호
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    • pp.112-123
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    • 1994
  • ${\gamma}$-Glutamyltransferase (GGT: E.C. 2.3.2.2.) is a glycoprotein enzyme which is involved in glutathione metabolism and amino acid transport through the plasma membrane. It is distributed widely in several organs including liver and kidney. Several isozymes of GGT have been reported and some of the isozymes may be associated with hepatocarcinogenesis. We have produced six monoclnal antibodies (mAbs) against GGT purified from the liver of 2-acetamidofluorene (AAF) treated rats. All of the six mAbs were obtained by immunizing mice with liver GGT Six hybridomas which produced anti-GGT Abs were extensively subcloned and injected into the peritoneal cavity of BALB/c mice to obtain large quantities of Abs. These mAbs were purified from ascites by ammonium sulfate precipitation and protein A sepharose CL-4B column chromatography. Using these mAbs we preformed enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunohistochemistry (IHC), and autoradiography (ARG) to study the distribution of GGT isozyme in tissue. The results indicate that GGT-mAb 1 is specific for the AAF treated liver GGT, GGT-mAb 5 for the normal liver GGT, and GGT-mAb 6 for the normal kindey GGT. These mAbs may be used to evaluate the distribution of GGT isozymes in different tissues.

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육제품에 첨가된 대두단백 정량을 위한 면역분석법 개발에 관한 연구: 대두단백 정량을 위한 항체생산 및 특성조사 (Development of Immunoassay Systems for the Assay of Soy Protein in Meat Products; Antibody Production and Properties for the Assay of Soy Protein)

  • 김천제;김종배;김병철;이승배;정성원;신현길;고원식
    • 한국식품과학회지
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    • 제24권3호
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    • pp.204-208
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    • 1992
  • 본 실험은 가공한 육제품에 첨가된 대두단백질(soy protein)을 정량하기 위한 면역분석(Immunoassay)법의 개발을 목적으로 실시하였다. Isolated soy protein(ISP)의 whole buffer extract(WBE) 분획을 SDS 처리 후 토끼에 주사하여 생산된 항혈청의 항체역가를 indirect ELISA법으로 조사하였을 시 1 : 10,000 이상에서도 반응을 나타내었다. 시료의 처리시 SDS의 최종농도가 0.03% 이상에서는 항원-항체 반응이 심각하게 저지되었으나, 0.02% 이하에서는 거의 영향을 미치지 않았다. 본 실험에 사용한 항체는 SDS로 변성된 항원(대두단백질)은 물론 SDS를 투석으로 제거한 재생항원(renatured antigen)과도 반응하였으나 그 정도는 변성항원에 비하여 약간 낮았다. 검정곡선(calibration curve)을 indirect competitive ELISA 법으로 작성하였을 시 ISP를 100 ng/100 ml까지 측정할 수 있는 감도(sensitivity)를 얻었다.

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