• Title/Summary/Keyword: antibodies

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Production and Characterization of Monoclonal Antibodies to Escherichia coli (ATCC 8739)

  • Yoo, Dohng-Hyun;Lee, Young-Ha;Jung, Jae-Deuk
    • Journal of Microbiology and Biotechnology
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    • v.5 no.6
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    • pp.353-358
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    • 1995
  • Escherichia coli causes intestinal and extraintestinal infections and has been an indicator of fecal pollution in water and food. BALB/c mouse was immunized by injection of somatic E. coli (ATCC 8739) cells to produce monoclonal antibodies. Splenocytes of mouse were fused with myeloma cells (Sp2/0-Ag14). Two hybridomas secreting monoclonal antibodies were established after being cloned. In SDS-PAGE analysis of E. coli antigens 37 protein profiles appeared from 14 kDa to 182 kDa. Western blot analysis using polyclonal antibodies demonstrated that protein antigens of 41 kDa, 38.2 kDa and 31.7 kDa were immunodominant. Monoclonal antibodies DY-CM1 and DY-CM2 recognized 31.7 kDa and 2.0 kDa antigens in Western blot analysis, respectely.

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Functionalization of Au surfaces with 4-(carboxymethyl)aniline and amine-terminated dendrimers for enhanced surface density of antibodies on immunosensor Au chips

  • Lee, Yongwoon;Ju, Youngwon;Kim, Joohoon
    • Analytical Science and Technology
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    • v.30 no.1
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    • pp.49-56
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    • 2017
  • Here, we demonstrate surface functionalization of Au chips with 4-(carboxymethyl)aniline (CMA) and amine-terminated polyamidoamine (PAMAM) dendrimers for immobilization of antibodies on the Au surfaces. Use of the functionalization strategy led to high surface density of the immobilized antibodies on the Au chips. Specifically, we found that the functionalization of Au chips with CMA and amine-terminated $6^{th}$ generation PAMAM dendrimers allowed immobilization of immunoglobulin (IgG) antibodies with high surface density, which is 5 times higher than that obtained with Au surfaces functionalized with CMA and ethylenediamine.

Anti-DNA Autoantibodies from on MRL/Ipr Mouse

  • Park, Jeong-Soo;Kim, Young-Tai;Lee, Chan-Hee;Youn, Jung-Koo;Jang, Young-Ju
    • Animal cells and systems
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    • v.2 no.3
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    • pp.371-375
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    • 1998
  • Twenty-one monoclonal anti-DNA autoantilndies were produced by fusing spleen cells from an autoimmune MRL/lpr mouse with SP2/0 myeloma cells. Hybridomas generated by the fusions were chosen for cloning on the basis of DNA binding by supernatant antibody. Each monoclonal antibody was purified to homogeneity and analyzed for the heavy and light chain isotypes and the binding specificity for single-stranded DNA, double-stranded DNA, and RNA. Sequence specificities and isoelectric points of the antibodies were also examined. All of the antibodies were lgG and tended to bind to both single-stranded and double-stranded DNA with a preference for the double-stranded form. Some of them also bound to RNA. Isoelectric points of the antibodies were shown to be high. The antibodies described in this report have characteristics of pathogenic anti-DNA antibodies.

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Effect of polymerization in inducing yolk antibodies (계란 항체의 생산에 있어서 polymerization의 효과)

  • Lee, Kyeong-Ae
    • Korean Journal of Human Ecology
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    • v.2 no.1
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    • pp.17-24
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    • 1993
  • Insulin polymer was used as an immunogen to elicit homogeneous anti-insulin yolk antibodies in a large scale. Insulin polymer prepared was heterogeneous mixture (MW=6,000-70,000). Insulin polymer showed stronger immunogenecity than insulin monomer. The affinity of yolk antibodies elicited with insulin polymer was slightly lower than that of yolk antibodies elicited with insulin monomer. The specificity of yolk antibodies obtained with insulin monomer and insulin polymer was directed mostly to native insulin.

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Production and Characterization of Monoclonal Antibodies to a Generic Hapten for-Class-Specific Determination of Organophosphorus Pesticides

  • Jang, Mi-Soon;Lee, Soo-Jung;Xue, Xiaoping;Kwon, Hyuk-Man;Ra, Choon-Sup;Lee, Yong-Tae;Chung, Tae-Wan
    • Bulletin of the Korean Chemical Society
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    • v.23 no.8
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    • pp.1116-1119
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    • 2002
  • Monoclonal antibodies have been generated against a generic hapten, ο,ο-diethyl ο-(5-carboxy-2-fluorophenyl) phosphorothioate, for the determination of organophosphorus (OP) pesticides in a class-specific manner. In an indirect competitive enzyme-linked immunosorbent assay (ELISA) format, employing a heterologous coating antigen, these monoclonal antibodies showed desirable properties for use in the class-specific determination, i.e., broad specificity and high sensitivity. The IC50 values of four commonly used ο,ο-diethyl OP pesticides were fairly uniform ranging from 0.1 to 0.3 ㎛/mL. The IC50 values of three ο,ο-dimethyl derivatives were between 0.3 and 1.4 ㎛/mL. These values, together with the limits of detection (LOD), were better, in terms of the specificity and sensitivity, compared with the values obtained previously with polyclonal antibodies.

Antisperm Antibodies in Infertile Males as Detected by Tray Agglutination Test and Indirect Immunobead Test (남성불임 환자에서 Tray Agglutination Test 및 Indirect Immunobead Test를 이용한 항정자항체에 관한 연구)

  • Kim, Cheol-Soo;Lee, Moo-Sang;Hong, Jae-Yup
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.2
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    • pp.153-158
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    • 1990
  • Sera from 95 infertile males males were assayed for antisperm antibodies using the Tray Agglutination Test(TAT) and indirect Immunobead Test(IBT). The correlation between antisperm antibodies and seminal analysis in infertile men was evaluated, and the TAT was compared with new indirect IBT. The results were obtained as follows. 1. Positive rate for antisperm antibodies was high in azoospermia, oligoasthenospermia, pyo-spermia, in order, Especially in obstructive azoospermia, the rate was the highest in both methods 2. Positive rates for antisperm antibodies in TAT was higher than indirect IBT. 3. Among the isotypes of the immunoglobulins, IgG were most prevalent. IgG and IgA were bound predominantly to the head and IgM predominantly to the tail up.

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Production of Monoclonal Antibodies Specific to FimA of Porphyromonas gingivalis and Their Inhibitory Activity on Bacterial Binding

  • Koh, Eun-Mi;Kim, Ju;Lee, Jin-Yong;Kim, Tae-Geum
    • IMMUNE NETWORK
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    • v.9 no.5
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    • pp.203-207
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    • 2009
  • Background: The FimA of Porphyromonas gingivalis is a crucial pathogenic component of the bacteria and has been implicated as a target for vaccine development against the periodontal diseases. Methods: In this study, the purified fimbriae (FimA subunit polymers) protein was used for immunization in their native form and B hybridoma clones producing antibodies specific to FimA were established. Results: The monoclonal antibodies prepared from selected two clones, designated #123 (IgG2b/ kappa) and #265 (IgG1/kappa), displayed different patterns of binding activity against the cognate antigen. Both antibodies reacted with conformational epitopes expressed by partially dissociated oligomers, but not with monomer as elucidated by Western blot analysis. Ascites fluid containing the monoclonal antibodies showed the inhibitory activity against P. gingivalis to saliva-coated hydroxyapatite beads, an in vitro model for the pellicle-coated tooth surface. Conclusion: These results suggest that the monoclonal antibodies could be used as vaccine material against the periodontal diseases through passive immunization.

Effects of Monoclonal Antibodies against Human Stathmin Combined with Paclitaxel on Proliferation of the QG-56 Human Lung Carcinoma Cell Line

  • Yuan, Shao-Fei;Chen, Wen-Jun;Zhu, Lin-Jia;Zheng, Wei-E.;Chen, Hua;Xiong, Jian-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2967-2971
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    • 2012
  • Objective: To explore whether monoclonal antibodies against stathmin and the chemotherapuetic agent paclitaxel have synergenic effects in inhibiting growth and inducing apoptosis in human QG-56 cells. Methods: QG-56 cells were treated with monoclonal antibodies against stathmin or paclitaxel alone or in combination, with untreated cells used as controls. After 24, 48, 72 and 96 hours the cell growth condition was observed under an inverted microscope and inhibition was studied by MTT assay; apoptosis was analyzed by flow cytometry. Results: The populations decreased and cell shape and size changed after the various treatments. Monoclonal antibodies against stathmin and paclitaxel used alone or incombination inhibited the proliferation of QG-56 cells, especially in combination with synergism (P<0.05). Combined treatment also resulted in a significantly higher apoptosis rate than in the other groups (P<0.05). Conclusions: Monoclonal antibodies against stathmin and paclitaxel used alone or in combination can inhibit proliferation of QG-56 cells and induce apoptosis when applied together. The observed synergistic effects may have important implications for clinical application.

Studies on the Characteristics of Anti-Zone Antibody I. Identification of Anti-Zona Antibody by Indirect ELISA and Indirect Immunofluorescence Test (항투명대 항체의 특성에 관한 연구 I. 간접효소면역분석법과 간접면역형광분석법에 의한 항투명대 항체의 확인)

  • 김은영;유영춘;박세필;정형민;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.107-114
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    • 1990
  • These experiments were undertaken as a basic study to develop immunocontraceptive vaccine and to understand the role of zona pellucidae in early fertilization process by identifying the monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae by indirect ELISA and indirect immunofluorescence test. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to zona was determined by indirect ELISA using solubilized porcine zona coated plates. Both monoclonal and rabbit polyclonal antibodies showed very high titers ; O.D at 1 : 12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. Rabbit anti-mouse zona pellucidae sera also reacted with porcine zona pellucidae. 2. By indirect immunofluorescence test strong fluorescences were observed on the egg treated with homologous and heterologous rabbit polyclonal antibodies and FITC lablled 2nd antibodies and found to crossreact strongly with the eggs from the pig and mouse. While weaken fluorescences were observed on the eggs treated with monoclonal antibodies.

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Detection of antibodies against infectious Borna disease virus -a comparison of three serological methods- (보르나병 바이러스 항체검출을 위한 연구 -세 가지 혈청진단법의 비교-)

  • Lee, Du-sik
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.57-61
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    • 1992
  • To determin the accuracy of serological methods in detecting Borna-disease(BD) viral antibodies, 273 experimentally infected rabbit sera were compared by using indirect immunofluorescence antibody test(IFA), serum neutralization test(SN) and enzyme-linked immunosorbent assay(ELISA). One hundred twenty-three serum samples had BD viral antibodies detected by IFA. CELISA antibodies to BD virus were also present in the same one hundred twenty-three serum samples. However, neutralization test antibodies to BD virus were present in 27 of the in rabbit serum samples. Neutralization test was sensitive in comparison with KFA and CELISA. In comparison with IFA, CELISA was both sensitive and specific in detecting BD viral antibodies. These results extend observations made with laboratory animals to the diagnosis of naturally infected animals.

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