• Title/Summary/Keyword: Polyclonal

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Polyclonal gammopathy related to renal bleeding in a peritoneal dialysis patient

  • Cho, Eun-Mi;Moon, Hye-Hyun;Hwang, Young-Ju;Lee, Seung-Jin;Ko, Cheol Woo;Cho, Min Hyun
    • Clinical and Experimental Pediatrics
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    • v.56 no.7
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    • pp.304-307
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    • 2013
  • Polyclonal gammopathy represents the diffuse activation of B cells and is usually related to inflammation or immune-related diseases. However, the mechanisms leading to polyclonal gammopathy are essentially speculative. Generally, infectious, inflammatory, or various other reactive processes may be indicated by the presence of a broad-based peak or band in the gamma region on serum protein electrophoresis results. A 15-year-old girl, who had been receiving peritoneal dialysis, presented with polyclonal gammopathy and massive gross hematuria. Renal artery embolization was performed, after which the continuous bleeding subsided and albumin-globulin dissociation resolved. This is a rare case of polyclonal gammopathy related to renal bleeding.

Effects of epitope sequence tandem repeat and proline incorporation on polyclonal antibody production against cytochrome 1A2 and 3A4

  • Ahn, Tae-Ho;Yun, Chul-Ho
    • BMB Reports
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    • v.42 no.7
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    • pp.418-420
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    • 2009
  • We describe a method for producing polyclonal antibodies against peptide antigen cytochrome P450 1A2 and 3A4 using a tandem repeat of the epitope region and incorporation of proline residue between the repeated sequences. An ELISA assay revealed more efficient generation of polyclonal antibodies to tandem repeat peptide antigens than mono-epitope peptides. The incorporation of proline residues further stimulated antibody production.

Development of an Indirect ELISA and Immunocapture RT-PCR for Lily Virus Detection

  • Kim, Jin Ha;Yoo, Ha Na;Bae, Eun Hye;Jung, Yong-Tae
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1776-1781
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    • 2012
  • Multiple viruses such as Lily symptomless virus (LSV), Lily mottle virus (LMoV), and cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gangwon, Chungnam, and Jeju provinces of Korea in 2008-2011. Coat protein (CP) genes of LSV and LMoV were amplified from collected samples by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into a pET21d(+) expression vector to generate recombinant CPs. The resulting carboxy-terminal His-tagged CPs were expressed in Escherichia coli strain BL21(DE3) by isopropyl-1-thio-${\beta}$-D-galactoside induction. The recombinant proteins were purified using Ni-NTA agarose beads, and the purified proteins were used as an immunogen to produce polyclonal antibodies in rabbits. The resulting polyclonal antisera recognized specifically LSV and LMoV from infected plant tissues in Western blotting assays. Indirect enzymelinked immunosorbent assay and immunocapture RT-PCR using these polyclonal antisera were developed for the sensitive, efficient, economic, and rapid detection of Lily viruses. These results suggest that large-scale bulb tests and economic detection of Lily viruses in epidemiological studies can be performed routinely using these polyclonal antisera.

Development and Characterization of Anti-gliadin Polyclonal Antibody in Wheat

  • Chang, Suk Joo;Hong, Byung Hee;Seo, Yang Weon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.4
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    • pp.339-344
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    • 1999
  • Immunological method has been applied in biochemical genetic analysis of seed storage proteins. We developed and characterized anti-gliadin polyclonal antibody (AGPab) specific to gliadin fractions whose quality and quantity were known to be associated with wheat end-use quality. Reactions of anti-gliadin polyclonal antibody (AGPab) to gliadin were linearly decreased as AGPab and antigen were diluted. Dot-blot and immunoblot assay showed that produced AGPab specifically reacted to gliadin and mainly $\alpha$-, $\beta$-, and ${\gamma}$-gliadin subunits. Enzyme-linked immuno- sorbent assay (ELISA) was applied for quantifi-cation of gliadins in Korean wheat cultivars and breeding lines by using AGPab. High reactions between AGPab and gliadins were found in wheat cultivars Olmil and Olgeurumil. Significant difference of optical densities for alcohol soluble proteins among crop species was found, as wheat showed the highest value (0.697) followed by rye (0.295), and barley (0.066).

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Comparison of IHNV Detection Limits by IMS-RT-PCR, Western Blot and ELISA

  • Kim Soo-Jin;Lee Eun-Young;Oh Myung-Joo;Choi Tae-Jin
    • Fisheries and Aquatic Sciences
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    • v.4 no.1
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    • pp.32-38
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    • 2001
  • Several molecular biological techniques have been used to detect virus rapidly and accurately, but these methods have limitations in the early stage of viral infection with very low concentration of virus. We compared the detection limits of IMS-PCR, Western blot and ELISA with infectious hematopoietic necrosis virus OHNV). Four antibodies, rabbit anti-IHNV polyclonal antibody, anti-IHNV nucleocapsid protein monoclonal antibody, anti-IHNV nucleocapsid protein polyclonal antibody, and anti-IHNV glycoprotein polyclonal antibody, were tested to find out the most effective antibody for each method. The detection limit with IMS- PCR was $2\times10^6$ pfu when the viral RNA was extracted before RT-PCR. In the western blot with rabbit anti­IHNV polyclonal antibody one pfu of virus could be detected. In ELISA, 10 pfu of virus particles were detected with the same antibody.

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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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Studies on the Characteristics of Anti-Zona Antibody II. Effect of Anti-Zona Antibody on Fertilization of Porcine and Mouse Eggs In Vitro (항투명대 항체의 특성에 관한 연구 II. 항투명대 항체가 돼지난자와 생쥐난자의 체외수정에 미치는 영향)

  • 김은영;박세필;정형민;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.115-124
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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 investigating the effect of monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae on the fertilization of porcine and mouse eggs in vitro. The results obtained in these experiments were summarized as follows : 1. Treatment of porcine and mouse eggs with undiluted anti-zona serum produced intense precipitation layer on the poricne and mouse zonae, respectively, thus resulting in the total inhibition of sperm adherence on surface of zona. 2. In vitro fertilization of eggs pre-treated with 0.3∼10% of various antibodies was examined, and resulting in that 5 and 10% of rabbit polyclonal antibodies to porcine zona inhibited completely both in vitro fertilization and polyspermy of porcine eggs while monoclonal to porcine zona and rabbit polyclonal antibody to mouse zona did not inhibit in vitro fertilization but monoclonal antibody reduced the rate of polyspermy compared to that of control group. Almost the same results were obtained in the study on the effect of anti-zona serum on in vitro fertilization of mouse eggs.

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Production of Polyclonal Antibodies against the Tegument of Sparganum (Plerocercoid of Spirometra mansoni) and Its Immunolocalization

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.48 no.2
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    • pp.183-185
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    • 2010
  • In a previous study, the author developed a method for separation of the tegument of spargana (plerocercoids of Spirometra mansoni) from the parenchyme using urea. The present study, as a next step, was performed to evaluate which molecules are present in the outer tegument. Two major proteins, 180 and 200 kDa, are present in the tegument and we could make polyclonal antibodies against these molecules. Their immunolocalization was processed and the outermost layer of the spargana showed strong positive staining. Conclusively, we could confirm that the 180 and 200 kDa molecules might be tightly bound membrane proteins in the tegument of spargana.

High-level Expression, Polyclonal Antibody Preparation and Bioinformatics Analysis of Bombyx mori Nucleopolyhedrovirus orf47 Encodes Protein

  • Wu, Chao;Guo, Zhongjian;Chen, Keping;Shen, Hongxing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.2
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    • pp.87-92
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    • 2008
  • Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.