• Title/Summary/Keyword: Antigen-antibody reaction

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Electronic Detection of Biomarkers by Si Field-Effect Transistor from Undiluted Sample Solutions with High Ionic Strengths

  • Ah, Chil-Seong;Kim, An-Soon;Kim, Wan-Joong;Park, Chan-Woo;Ahn, Chang-Geun;Yang, Jong-Heon;Baek, In-Bok;Kim, Tae-Youb;Sung, Gun-Yong
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1561-1567
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    • 2010
  • In this study, we have developed a new detection method using Si field effect transistor (FET)-type biosensors, which enables the direct monitoring of antigen-antibody binding within very high-ionic-strength solutions such as 1$\times$PBS and human serum. In the new method, as no additional dilution or desalting processes are required, the FET-type biosensors can be more suitable for ultrasensitive and real-time analysis of raw sample solutions. The new detection scheme is based on the observation that the strength of antigen-antibody-specific binding is significantly influenced by the ionic strength of the reaction solutions. For a prostate specific antigen (PSA), in some conditions, the binding reaction between PSA and anti-PSA in a low-ionic strength reaction solution such as 10 ${\mu}M$ phosphate buffer is weak (reversible), while that in high-ionic strength reaction solutions such as 1$\times$PBS or human serum is strong.

Detection of Hepatitis C Virus by RIA, Recombinant Immunoblot Assay and Nested RT-PCR (RIA, Recombinant Immunoblot Assay 및 Nested RT-PCR에 의한 C형 간염바이러스 검출)

  • Kim, Jae-Soo;Kim, Jong-Wan;Lee, Yun-Tai
    • The Journal of Korean Society of Virology
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    • v.30 no.2
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    • pp.151-159
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    • 2000
  • Hepatitis C virus (HCV) is one of the important human pathogen that can cause acute and chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Recently, the third generation radiation immuno assay (RIA) method has been developed as a very sensitive test to detect anti-HCV antibody. However, false positive is the problem with RIA test. To solve this the RIA results were compared to those of 5-antigen recombinant immunoblot assay (5-RIBA) and reverse transcription-polymerase chain reaction (RT-PCR). Among 12,767 serum samples tested from clinic visitors, total 275 (2.2%) samples were antibody positive by RIA. RIBA was performed with 148 RIA positives cases but among them was shown eighty five was antibody positive and sixty three (42.6%) was negative result. However, nested RT-PCR test was shown also carried out with 43 positive, 6 intermediates and 25 negatives of RIBA. As a result of the nested RT-PCR results, HCV antigen were detected in RIBA positive, 33.3% (2/6) RIBA intermediate and 12% (3/25). Clinical syndrome of all 148 patients as a with chronic active hepatitis (46.0%), cirrhosis (18.9%), hepatocellular carcinoma (8.1%) and others (27.0%) and they were positive in reaction by RIA test. But RIBA positive patients with 34.9% of chronic active hepatitis, 18.6% of cirrhosis, 4.6% of hepatocellular carcinoma and 41.9% of others were detected to be positive case by nested RT-PCR.

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Variation of antigenicity and serological reaction to Pneumocystis carinii in Korea

  • Park, Hyun-Young;Lee, Soo-Ung;Chae, Seong-Wan;Huh, Sun;Yu, Jae-Ran;Kim, Jin;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • v.37 no.2
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    • pp.109-115
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    • 1999
  • The present study observed the variation of antigenicity of Pneumocystis carinii and serum IgG antibody reastion to the antigens from different localtities in Korea. Antigens of rat P,carinii and sera of inhabitants were collected at Chunchon, Chungju, Kwangju and Seoul during 1995-1996. Enzyme-Linked Immunosorbent Assay and immunoblot were used for immune reaction. Absorbance of 1,294 human sera ranged between 0.01and 0.93. Sera from Chunchon showed higher absorbances than those from other areas. Immunoblotting revealed IgG antibody reactions to 116,100, and 45-55 kDa antigenic bands of rat P.carinii, but the frequencies of positive reaction to individual bands were variable by localities. Total 62.6% reacted to 100kDa band and 32.0% to 45-55 kDa bands. For the reaction to 116kda, the reaction rate was 60.0% of the sera showed the reaction to 116kda band while 37.7% reacted to 100kda, the reaction rate was 60.0% to 82.6% by localities. It is found that the reaction rates of the human sera to rat P.carinii antigen are variable according to the localities. Also, the high molecular antigen of 116kDa of rat P.carinii is the most frequent antigenic band reaction to human sera.

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Prevalence of dog erythrocyte antigen (DEA) 1 among Jindo dogs (진돗개에서 개적혈구항원(dog erythrocyte antigen) 1 혈액형 조사)

  • Lee, Jeong-Chi
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.271-276
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    • 2018
  • This study was conducted to investigate the prevalence of dog erythrocyte antigen (DEA) 1 with DEA 1.1 and DEA 1.2 on 122 Jindo dogs (29 males, 93 females) from 2014 to 2015 using a monoclonal antibody card kit (blood typing card kit, Korea Animal Blood Bank Inc., South Korea). Among the tested dogs, 14.8% (18/122) were positive for the DEA 1.1 antigen and 85.2% (104/122) were positive for the DEA 1.2 antigen. The prevalence of positive types for the DEA 1.2 antigen was significantly higher than the DEA 1.1 antigen (P<0.01). The prevalence of positive types for the DEA 1.1 antigen was higher in white-haired Jindo dogs than yellow-haired dogs (P<0.05). However, there was no gender difference in the prevalence of the DEA 1.1 antigen (P=0.665). The incidence of sensitization after the first transfusion without blood group test was 12.6% and the incidence of acute hemolytic transfusion reaction after the second transfusion in the same immunized dogs was 1.6%. Therefore, the blood group test for the DEA 1 antigen should be performed for Jindo dogs to ensure safe and effective transfusion therapy and further studies remain to be conducted for other DEAs among Jindo dogs.

Application of monoclonal antibody to develop diagnostic techniques for infectious bovine rhinotracheitis virus. II. Diagnosis of infectious bovine rhinotracheitis by using monoclonal antibody (소 전염성비기관염(傳染性鼻氣管炎) 바이러스에 대한 monoclonal antibody 생산(生産)과 진단법(診斷法) 개발 II. Monoclonal antibody를 이용한 소 전염성비기관염(傳染性鼻氣管炎)의 진단(診斷))

  • Jun, Moo-hyung;Kim, Duck-hwan;An, Soo-hwan;Lee, Jung-bok;Min, Won-gi
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.27-35
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    • 1989
  • To develop more specific and sensitive diagnostic methods for infectious bovine rhinotracheitis, 7-C-2 monoclonal antibody specific to polypeptides of infectious bovine rhinotracheitis virus (IBRV) was applied in indirect immunofluorescence antibody assay (IFA), indirect immunoperoxidase assay(IPA) and radial immunodiffusion enzyme assay (RIDEA). It was found that IBRV infected in MDBK cells could be detected as early as 8 hours post infection by IFA, and that IFA was more rapid and specific to identify IBRV antigen than IPA. The diagnostic efficacy of RIDEA and SN test was studied with 88 bovine sera. It was evident that RIDEA could eliminate the false positive reaction encountered in serum neutralization(SN) test, being more rapid and sensitive than the latter. Highly significant correlation coefficiency (r=0.76, p<0.01) was evaluated between the titers of sera and the diameters of RIDEA. Tracheal membranes and sera collected from 96 slaughtered cattle with lesions in respiratory organs were examined to detect IBRV antigen and antibody by IFA, RIDEA and SN test. It was presented that positive rates were 32.3% in IFA, 20.8% in RIDEA and 21.9% in SN test, and that coincidence rate between RIDEA and SN test were 100% in positive sera and 98.7% in negative sera. In conclusion, it was assumed that application of monoclonal antibody could improve the diagnostic efficacy of IBR by enhancing sensitivity and specificity of IPA, IFA and RIDEA.

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The Principles and Practices of Immunocytochemical method in Light Microscopic Level (광학 현미경적 수준에서의 면역조직화학적 방법의 원리 및 실제)

  • Kim Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.3 no.1
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    • pp.229-250
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    • 1991
  • The study was carried out to investigate and review the principles and practices of immunocytochemical method in light microscopic level. The results were as follows. 1. Immunocytochemistry is the method to search out the intracellular position of the specific meterials using antigen -antibody reaction. 2. The chief items in immunocytochemistry are antigen, antibody and chromogen. 3. The identifical fixation is cardiac perfusion fixation. 4. The tissue slides must be prepared by vibratomy. 5. All stainings are carried out with free floating staining method. 6. There are polyclonal and monoclonal antibodies used in immunocytochemistry.

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Immunohistochemical application of anti-proliferating cell nuclear antigen antibody on the development of uterus following sex hormone administration in rats (성 hormone이 rat 자궁 발달에 미치는 영향에 대한 proliferating cell nuclear antigen 항체의 면역조직학적 응용)

  • Koh, Phil-ok;Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.37 no.2
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    • pp.281-289
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    • 1997
  • The study was designed to investigate the effects of progesterone and estrogen on the uterus of rats by immunohistochemical methods using Proliferating Cell Nuclear Antigen (PCNA) antibody. Eighteen female rats(Wistar), weighing initially about 300g, were ovariectomized. These rats were divided into four groups, progesterone-treated group, estrogen-treated group, estrogen+progesterone-treated group, and control group, progesterone-treated group was injected with 1mg of progesterone per rat per day for 2 days and estrogen-treated group with $20{\mu}g$ of $17{\beta}-estradiol$ for 3 days and estrogen+progesterone-treated group with $17{\beta}-estrdiol$ for 3 days and then with progesterone for 2 days as above. In gross findings, the uteri were markedly hypertrophied by estrogen treatment but were not affect in size by progesterone treatment. Immunohistochemical investigation was performed on the cell types with higher appearance of PCNA positive reaction cells in four groups. The groups with higher appearance of the stromal cells were ordered as estrogen-treated group, progesterone-treated group, estrogen+progesterone-treated group, and control group. The muscle cells were ordered as progesterone-treated group, estrogen-treated group, estrogen+progesterone-treated group, and control group. Positive reaction cells of the stromal cells were total 4.6 times higher than those of muscle cells. Therefore, the affect of the hypertrophy on the uterus by estrogen was larger than those of progesterone and affect on the uterus by stromal cells were larger than those of muscle cells. The group with more PCNA positive reaction cells of luminal epithelial cells were ordered as control group, progesterone-treated group, estrogen+progesterone-treated group, and estrogen-treated group, and glandular epithelial cells were ordered as estrogen+progesterone-treated group, progesterone-treated group, control group, and estrogen-treated group. It was suggested that estrogen and progesterone did not affect on the proliferating cells of luminal epithelial cells and affection of progesterone on the development of glandular epithelial cell was larger than that of estrogen.

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Detection of Protein Molecules by Electrical Current Response Using Two-Electrode Method

  • Lyu, Hong-Kun;Woo, Sung-Ho;Han, Yoon-Soo;Lee, Hee-Ho;Shin, Jang-Kyoo
    • Journal of Sensor Science and Technology
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    • v.20 no.2
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    • pp.90-95
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    • 2011
  • In order to protect human lives from disease, various biosensors having the potential to analyze a variety of biomolecules have been utilized. Biosensors constitute one of the most promising ways to monitor and detect various biomolecules corresponding to diseases. In this study, we demonstrate that the reaction of streptavidin molecules with biotin on a gold electrode can be detected using the twoelectrode method with a gold electrode and a platinum reference electrode. We also show the characteristics of the electrical current response. While detecting 2-${\mu}M$ streptavidin molecules dissolved in phosphate buffered saline(PBS) solution, we found that an analytical biosensor can operate on the principle of detecting an antigen-antibody reaction event of protein molecules using the two-electrode method. We think that the "potential step" method might be useful to detect the occurrence of any antigen-antibody reactions and can be combined with other devices or ICs such as BJTs, MOSFETs, and OP-amps for the detection of biomolecules of diseases.

Cell-Specific Targeting of Texas Red with Anti-Ep-CAM Antibody

  • Lee, Soo-Chul;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.123-127
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    • 2005
  • The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

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High $T_c$ SQUID system for biological immunoassays

  • Enpuku, K.
    • 한국초전도학회:학술대회논문집
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    • v.10
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    • pp.7-7
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    • 2000
  • A high $T_c$ SQUID system is developed for the application to biological immunoassay. In this application, magnetic nanoparticles are used as magnetic markers to perform immunoassay, i.e., to detect binding reaction between an antigen and its antibody. The antibody is labeled with ${\gamma}-Fe_2O_3\;(or\;Fe_3O_4)$ nanoparticles, and the binding reaction can be magnetically detected by measuring the magnetic field from the nanoparticles. Design and set up of the system is described. The system consists of (1) SQUID magnetometer or gradiometer made of 30-deg. bicrystal junctions, (2) field and compensation coils to apply the magnetic field of about 1 mT, (3) special Dewar to realize a 2 mm-distance between the SQUID and the sample, (4) two layers of cylindrical shielding to reduce the extemal magnetic noise to about 1/100, and (5) an electric slider to move the sample with a speed of 10 mm/sec. The sensitivity of the system is studied in terms of detectable magnetic flux. For the measurement bandwidth from 0.2 Hz to 10 Hz, minimum-detectable amplitude of the magnetic flux is $0.8\;m\;{\Phi}_o$ and $0.25\;m{\Phi}_o$ for the magnetometer and the gradiometer, respectively, when the magnetic field of 1 mT is applied. The difference between them is due to the residual environmental noise, and the applied magnetic field does not increase the system noise. The corresponding weight of the magnetic markers is 1 ng and 310 pg, respectively. An experiment is also conducted to measure antigen-antibody reaction with the present system. It is shown that the sensitivity of the present system is 10 times better than that of the conventional method using an optical marker. A one order of magnitude improvement of sensitivity will be realized by the sophistication of the present system.

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