• Title/Summary/Keyword: Antibody specificity

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Production of Egg Yolk Antibody (IgY) Against Human Placental DNA-Dependent RNA Polymerase II

  • Lee, Yoon-Ik;Surzycki, Stefan S.;Lee, Young-Ik
    • BMB Reports
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    • v.28 no.1
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    • pp.27-32
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    • 1995
  • Polyclonal antibodies against human DNA-dependent RNA polymerase II (HPP II) were generated from chicken egg yolk after immunization with RNA polymerase II as an antigen. The antibodies from egg yolk (IgY) were purified and characterized. IgY showed a specificity against DNA-dependent RNA polymerase II, and was a polyclonal antibody against 12 subunits of polymerase II. An amount of 0.35 mg of IgY was obtained freman HPP II-Sepharose affinity column using 10 eggs from a chicken immunized against RNA polymerase II as an antigen. These antibodies can be used for isolating the genes for RNA polymerase II components, and for in vitro transcription assays using HP-RNA polymerase II.

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Production of Polyclonal Antibodies Specific to Porcine Adipocyte Plasma Membrane Proteins in Sheep (면양을 이용한 돼지 지방제포 원형질막 단백질 특이 항체의 생산)

  • 최창본;이명진;권은진
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.57-63
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    • 1998
  • The objectives of this study were to produce polyclonal antibody to adipocyte plasma membrane (APM) proteins isolated from pig, and to investigate its tissue specificity. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver and spleen were isolated using a self-forming Percoll gradient. Sheep (40kg) was immunized three times at three week interval with the purified APM proteins. Blood was taken from non-immunized sheep (NS) and from immunized sheep at 10 (AS-1), 12 (AS-2), and 14 (AS-3) days after the third immunization. Antisera titers and cross-reactivity against other tissues were determined by enzyme-linked immunosorbent assay (ELISA). Antisera reacted strongly to APM proteins showing detectable amounts of antibody at 1:81,000 dilution. And antisera showed much stronger reactivity to APM proteins than any other tissue plasma membrane proteins. Furthermore, tissue specificity of antisera against APM was reconfirmed by immunoblotting using anti-sheep immunoglobulin G-horseradish peroxidase conjugate as a secondary antibody Antisera to APM proteins showed adipocyte specificity compared with other tissues. In conclusion, polyclonal antibody against APM proteins isolated from pig was developed successfully in our laboratory, and these antisera showed tissue specificity with APM.

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Detection of antibodies and antigens of Aujeszky's disease virus using dot-immunoassay (Dot-immunoassay를 이용한 Aujeszky's disease virus항원 및 항체 검출)

  • Jun, Moo-hyung;Cho, Young-sung;Chang, Kyung-soo
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.529-536
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    • 1994
  • To establish more specific and simple diagnostic methods for detection of the antibodies and antigens of Aujeszky's disease virus(ADV), we designed indirect dot-immunoassay(IDI) and double sandwich dotimmunoassay(DSDI) using the solid phases of nitrocellolose paper and polystyrene plate. The diagnostic efficacy of these methods was investigated. As the sensitivity of IDI was tested by various virus concentration, the specimens with the virus titer above $10^{4.0}TCID_{50}/0.2ml$ showed positive reaction, but that below $10^{1.0}TCID_{50}/ml$ revealed negative. Tonsil emulsion at the virus titer of $10^{4.5}TCID_{50}/0.2ml$ showed the highest sensitivity as diluted by 1/100. In detection of ADV antigens from the various tissues of the rats and pigs infected with ADV, IDI using monoclonal antibody showed the higher specificity as compared with IDI using polyclonal antibody and virus isolation method. The efficacy of the DSDI for detection of ADV antibody was compared with other tests. The sensitivity of DSDI was higher than virus neutralization(VN) and agar gel immunodiffusion test(AGID). Meanwhile, specificity of DSDI was lower than AGID, but similar to IDEA. In comparison with VN test, DSDI showed 96.9% agreement to VN test that is the highest of three tests. In general, application of polyclonal antibody in both tests caused the higher sensitivity but the lower specificty.

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Diagnostic Value of Immunoglobulin G Anti-Deamidated Gliadin Peptide Antibody for Diagnosis of Pediatric Celiac Disease: A Study from Shiraz, Iran

  • Anbardar, Mohammad Hossein;Haghighi, Fatemeh Golbon;Honar, Naser;Zahmatkeshan, Mozhgan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.25 no.4
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    • pp.312-320
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    • 2022
  • Purpose: Screening serologic tests are important tools for the diagnosis of celiac disease (CD). Immunoglobulin (Ig)G anti-deamidated gliadin peptide (anti-DGP) is a relatively new autoantibody thought to have good diagnostic accuracy, comparable to that of anti-tissue transglutaminase (anti-tTG) antibody. Methods: Pediatric patients (n=86) with a clinical suspicion of CD were included. Duodenal biopsy, anti-tTG, and IgG anti-DGP antibody tests were performed. The patients were divided into CD and control groups based on the pathological evaluation of duodenal biopsies. The diagnostic accuracy of serological tests was determined. Results: IgA anti-tTG and IgG anti-DGP antibodies were positive in 86.3% and 95.4% of patients, respectively. The sensitivity, specificity, and diagnostic accuracy of the IgA anti-tTG test were 86.3%, 50.0%, and 68.6%, respectively, and those of the IgG anti-DGP test were 95.4%, 85.7%, and 90.7%, respectively. The area under the receiver operating characteristic (ROC) curve was 0.84 (95% confidence interval [CI], 0.74-0.91) for IgA anti-tTG test and 0.93 (95% CI, 0.86-0.97) for IgG anti-DGP test. The comparison of IgA anti-tTG and IgG anti-DGP ROC curves showed a higher sensitivity and specificity of the IgG anti-DGP test. Conclusion: IgG anti-DGP is a reliable serological test for CD diagnosis in children. High tTG and DGP titers in the serum are suggestive of severe duodenal atrophy. The combined use of IgA anti-tTG and IgG anti-DGP tests for the initial screening of CD can improve diagnostic sensitivity.

Demonstration of TCM-9 Monoclonal Antibody in Follicular Neoplasm of Thyroid (갑상선의 여포상 종양의 감별에 있어서 TCM-9의 발현양상)

  • Kim, Yun-Jung;Shim, Jung-Weon;Ahn, Hye-Kyung;Park, Young-Euy
    • The Korean Journal of Cytopathology
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    • v.7 no.2
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    • pp.134-137
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    • 1996
  • Monoclonal antibody(TCM-9) against human thyroid cancers have been studied by screening with human thyroid cancers, normal and benign thyroid tissue, and normal human serum protein. A monoclonal antibody(TCM-9) that is known to have strong specificity for human thyroid cancer but not for Graves' disease, adenoma or normal thyroid does not bind to native or mature human thyroglobulin(Tg). We used to TCM-9 antibody by immunohistochemical staining on 5 follicular cancer, 2 follicular adenoma, 1 follicular neoplasm with suspicious invasion, 2 papillary cancer to ascertain being of help in differentiation between follicular carcinoma and adenoma. Reactivity of TCM-9 was observed in follicular carcinoma and papillary carcinoma but not observed in follicular adenoma. Thus TCM-9 is a novel monoclonal antibody against the thyroid cancer.

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Detection of IgG and IsM Antibodies with Immunofluoreseent Antibody Technique in Buman Trichomoniasis (질트리코모나스증에서 간접형광항체법을 이용한 혈청내 항질트리코모나스 IgG 및 IgM 항체의 측정)

  • 윤경찬;김경민;안명희;민득영;차동수
    • Parasites, Hosts and Diseases
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    • v.25 no.1
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    • pp.7-12
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    • 1987
  • The indirect fluorescent antibody(IFA) test was used to detect serum IgG and IgM antibodies to Trichomonas vaginalis in 31 vaginal trichomoniasis, 7 candidiasis and in 20 non-infected healthy women with antigen prepared from axonic culture of Trichomenas vaginalis isolated from vulvovaginitis patient. The results were as follows: 1. In 31 vaginal trichomoniasis the positive reactions of IgG antibody were 27 in the 1/8 dilution or higher and :l in the 1/4 dilution whereas in healthy women the reaction showed significantly low as in the 1/4 dilution or below. 2. The sensitivity and specificity of IFA test for IgG antibody to trichomonad antigen in this study were 87.1% and 100%, respectively. 3. No significant difference of IgM antibody levels between vaginal trichomoniasis and healthy women was observed. 4. No relation between the levels of IgG and IsM antibodies to trichomonad antigen by IFA test was observed. 5. No relation between the time lapse and the level of serum IgG antibodies in IFA test of vaginal trichomoniasis was regarded. In conclusion the present study suggests that IFA test in trichomoniasis could be a useful tool for detection of anti-trichomonad IgG antibodies and applicable as an immunodiagnostic method.

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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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Immunocytolocalization of Cell Wall Peroxidase and Other Wall Antigens from Maize Seedlings

  • Kim, Sung-Ha
    • Journal of Plant Biology
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    • v.39 no.2
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    • pp.99-105
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    • 1996
  • Immunocytochemistry utilizes the specificity of the antigen-antibody reaction to localize specific antigens in cells or cellular organelles. Here we report the use of monoclonal antibodies, in conjunction with gold-labeled second antibodies to study the ultrastructural localization and tissue distribution of the Mr 98, 000 anionic peroxidase and other wall antigens. The antibody specific for this wall peroxidase, mWP3, labeled mainly the cell wall area. At the tissue level, the Mr 98, 000 peroxidase is located predominantly in the leaf mesophyll, internal coleoptile and sieve elements, but not in the root, as assayed with these procedures. The coleoptile walls were less heavily stained than the walls of leaf mesophyll cells. At the subcellular level, it is localized mainly in intercellular regions of the cell walls. A similar staining pattern was revealed by mWP19, one of anti-$\beta$ glucosidase antibody, though it looked less heavily stained than one with mWP3. In order to serve as a control wall staining using IgM monoclonal antibodies, mWP18 was used. Most of the label is localized over wall regions of cells of the young leaf mesophyll and coleoptile.

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A carbohydrate antigen of Clonorchis sinensis recognized by a species-specific monolonal antibody (종특이 단세포군항체에 반응하는 간흡충의 당질항원 한 가지)

  • 용태순;이종석
    • Parasites, Hosts and Diseases
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    • v.34 no.4
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    • pp.279-282
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    • 1996
  • The enzyme-linked immunosorbent assay (ELISA)-inhibition test using a Clonorchis sinensis species-specific mouse monoclonal antibody (MAb) , CsHyb 0605-23, showed increased specificity over the conventional ELISA used for serodiagnosis of clonorchiasis. To characterize the corresponding antigen further, the MAb was tested against polysaccharide, protein and glycolipid fractions obtained from a crude extract of C. slnensis adult worms, using chloroform, methanol and phenol extractions. Only the polysaccharide fraction was recognized by the mb among those fractions. Mild oxidation of the antigen with sodium periodate showed decreased reactivity against the MAb. We concluded that the antigen and antigenic determinants recognized by the MAb are carbohydrates.

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Assay of Cellobiohydrolnse by Column Single Immunodiffusion and Enzyme tinted Immunosorbent Assay (면역화학적 방법에 의한 Cellobiohydrolase 정량)

  • 오태광;고영희;김정일;박관희
    • Microbiology and Biotechnology Letters
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    • v.16 no.3
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    • pp.226-230
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    • 1988
  • Antibody against cellobiohydrolase purified from Trichoderma viride had been obtained by injection to rabbit. The antibody had a high specificity against the cellobiohydroase evidienced by absence of immunological reaction to other isozymes from Trichoderma viride. Assay limit of cellobiohydrolase was 1-10 $\mu\textrm{g}$ by column single immunodiffusion and by enzyme linked immunosorbent assay, it was 10-140 ng and 100-1200 pg when the dilution of antibody was 10$^{-6}$ and 10$^{-5}$, respectively.

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