• Title/Summary/Keyword: 다클론

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Reactivity of the Antibodies against Purified Carp Vitellogenin and a Synthetic Vitellogenin Peptide (정제 잉어 Vitellogenin과 합성 Vitellogenin 펩타이드에 대한 항체의 반응성)

  • Moon, Dae-Kyung;Kim, Nam-Soo;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.196-201
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    • 2006
  • Vitellogenin, which is found in the serum of female and male fishes exposed to environmental endocrine disrupter or estrogen hormone, is used as a biomarker for environmental contamination with an endocrine disrupter. In order to produce antibody against vitellogenin, a synthetic peptide for partial vitellogenin was injected into rabbits. In addition, by using ion exchange chromatography on DE-52, vitellogenin was purified from the serum of carp induced with $17{\beta}$-estradiol. Polyclonal antibody against purified vitellogenin reacted well with vitellogenin in the serum of carp induced with $17{\beta}$-estradiol and the serum of female carp, whereas polyclonal antibody against the vitellogenin peptide did not react with proteins in those samples. This may indicate that vitellogenin proteins, covalently modified largely, could not be detected by Western blotting with the polyclonal antibody against the synthetic vitellogenin peptide.

Immunoblotting Assay for Glyphosate-tolerant Genetically Modified Soybean in Soybean Products (식품 중 제초제내성 유전자재조합 콩의 immunoblotting 분석)

  • Son, Dae-Yeul;Moon, Jung-Hee;Ahn, Kang-Mo;Shon, Dong-Hwa;Lee, Kwang-Shin;Sim, Hee-Young;Han, Young-Shin;Lee, Sang-Il
    • Korean Journal of Food Science and Technology
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    • v.36 no.3
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    • pp.369-374
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    • 2004
  • Genetically modified (GM) soybean Roundup Ready carries Agrobacterium sp. CP4 gene, which expresses 5-enolpyruvylshikimate-3-phosphate synthase (CP4EPSPS). CP4EPSPS in GM soybeans and soybean curds was screened using CP4EPSPS-specific polyclonal and monoclonal antibodies (pab and mab, respectively) by immunoblotting. Isolated recombinant CP4EPSPS was detected at detection limits of $0.006\;and\;0.0006{\mu}g$, whereas those of CP4EPSPS expressed in GM soybean were $0.001\;and\;0.0001{\mu}g$g, using mab and pab, respectively. From nine screened soybean curds, two had positive results with pab Immunoblotting method with pab and mab developed in this study could be applied to screen glyphosate-tolerant GM soybeans in soybean products.

Vitellogenin ELISA System Based on Monoclonal and Polyclonal Antibodies against Vitellin of Floating Goby (Chaenogobius annularis) (꾹저구(Chaenogobius annularis)의 난황단백질에 대한 다클론 항체와 단글론 항체을 이용한 Vitellogenin ELISA System)

  • Kang, Bong-Jung;Jung, Jee-Hyun;Lee, Je-Yong;Kim, Myung-Hee;Han, Chang-Hee
    • Development and Reproduction
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    • v.9 no.2
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    • pp.135-142
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    • 2005
  • Vitellogenins(VTGs) are the precursor of egg-yolk proteins in most oviparous species from invertebrates to vertebrates. In oviparose vertebrates, VTGs are synthesized in the liver and transported through the blood to oocytes. In female fish, concentrations of plasma VTG increase rapidly at onset of vitellogenesis in the normal reproductive cycle. Male fishes also possess the gene for VTG, but plasma concentrations of the protein typically remain small, presumably due to low levels of endogenous estrogens. However, exposure of males to exogenous estrogenic mimics can result elevated. Therefore, the VTG in fish can be used as a useful biomarker for appropriate tools of endocrine disrupting compounds effects. In this studies, we prepared the test methods that can measure the plasma VTG level in the gobies that live in polluted area with mimic estrogen. For the purpose, we purified VTG of floating goby(Chaenogobius annularis) and prepared specific monoclonal and polyclonal antisera to yolk protein, then developed a sandwich competitive ELISA system for measurement of plasma VTG levels. Validation for the ELISA system using monoclonal and polyclonal antibodies against VTG was tested. The absorbance curve of serial dilutions of serum from vitellogenic female was paralleled to the standard curve of VTG, but normal male was not paralleled. The developed sandwich ELISA system was measured for VTG levels in plasma of common goby(Acanthogobius flaviman) and javeline goby(A. hasta) as well as in plasma of floating goby(C. annularis).

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Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocyte for Reducing Body Fat of Rat (흰쥐 체지방 감소를 위한 지방부위별 특이 다클론 항체 개발 연구)

  • Choi, Chang-Weon;Park, Dong-Woo;Jo, Ik-Hwan;Oh, Young-Kyoon;Chang, Sun-Sik;Choi, Nag-Jin;Kam, Dong-Keun;Baek, Kyoung-Hoon
    • Journal of agriculture & life science
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    • v.46 no.5
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    • pp.47-55
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    • 2012
  • Polyclonal antisera against regional (abdominal and subcutaneous) fats were developed to reduce body fat in rats. Isolation and culture of abdominal and subcutaneous adipocytes of rats were performed for analyzing lactate dehydrogenase (LDH) concentration. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas both of antisera against abdominal (AAb) and subcutaneous adipocyte plasma membrane proteins (SAb) had relatively strong reactivity till the level of 1:128,000 dilutions. Compared with regional fats, extremely low reactivities of AAb and SAb were detected with PMP of the organs (p<0.001). Both AAb and SAb were most strongly reacted with each adipocyte plasma memebrane proteins and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum based on LDH analysis. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte plasma membrane proteins are well developed and have safety in cross-reactivities with body organs.

Polyclonal Antibody against Paenibacillus larvae and its Application (Paenibacillus larvae에 대한 다클론 항체 및 그 응용)

  • 백경찬;양옥순;정규회;윤병수
    • Korean journal of applied entomology
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    • v.41 no.1
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    • pp.49-53
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    • 2002
  • Paenibacillus larvae is a gram-positive, spore-forming bacterium that is etiological agent for american foulbrood disease (AFB), which is the most severe disease in honey bee. To detect P. larvae from infected honeybee-comb or larvae, polyclonal antibody against whole bacterium was produced from guineapig and its specificity was evaluated. After optimization of ELISA-based detection system using these antibodies, a number of different P. larvae strains were analysed. Polyclonal antibody against P. larvae ATCC 25747 showed high affinity to most strains of P. larvae including P. larvae. strain ATCC 9545 (type strain), ATCC 25747 and other korean strain, SJl5 but exhibited no cross-reaction with other bacterial species. Additionally, this type of ELISA system was used for the detection of AFB in field-application The results have shown that this antibody could be useful for the rapid identification and monitoring of P. larvae in honeybee-comb.

Comparison of Growth Characteristics Between 5-year-old emblings Derived form Somatic Embryos and Seedlings in Liriodendron tulipifera (백합나무 5년생 체세포배 유래 클론배양묘 및 실생묘 간의 생장특성 비교)

  • Kim, Yong Wook;Moon, Heung Kyu
    • Journal of Korean Society of Forest Science
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    • v.101 no.4
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    • pp.613-618
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    • 2012
  • The 5-year-old seedlings and emblings which regenerated from somatic embryos were compared to the height, DBH, foliar characteristics, content of chlorophyll (chlorophyll a, b and total chlorophyll), carotenoid and leaf microstructure in Liriodendron tulipifera. In comparison of height and DBH (diameter at breast height), no significant differences were found in height (seedling, 3.8 m; embling, 3.87 m) and DBH (seedling, 12.09 cm; embling, 12.53 cm). The emblings and seedlings were similar in values of length (seedling, 108.11 mm, embling, 113.59 mm), width (seedling, 149.1 mm; embling, 167.71 mm), surface area (seedling, $119.92mm^2$; embling, $164.43mm^2$), fresh weight (seedling, 2.1 g; embling, 2.62 g) of leaf, and length (seedling, 81.49 mm; embling, 98.41 mm) and thickness (seedling, 1.66 mm; embling, 1.98 mm) of petiole. In case of chlorophyll content in the leaves, the chlorophyll a (seedlings, $11.2{\mu}g/g$; emblings, $13.2{\mu}g/g$), b (seedlings, $4.8{\mu}g/g$; emblings, $5.4{\mu}g/g$) and total content were higher in emblings ($930.2{\mu}g/g$) than seedlings ($800.1{\mu}g/g$), however, content of carotenoid (seedlings, $260.3{\mu}g/g$; embling, $265.2{\mu}g/g$) showed similar in both plants. Leaves of emblings had a similar pattern of histological structure (palisade or sponge parenchyma) to that of seedlings leaves. Therefore, the results showed that there were no remarkable growth differences when compared with 5-year-emblings and seedlings of yellow poplar.

Allergenicity Change of Soybean Proteins by Thermal Treatment (열처리에 따른 콩 단백질 Allergenicity 변화)

  • Son, Dae-Yeul;Lee, Bo-Ryun;Shon, Dong-Wha;Lee, Kwang-Shin;Ahn, Kang-Mo;Nam, Sung-Yeon;Lee, Sang-Il
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.959-963
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    • 2000
  • Soy bean is one of the most common food material to cause food hypersensitivity reactions in Korea. In this study we have investigated the effect of heating on antigenicity and allergenicity change of soybeans by using immunoblotting and ELISA methods with serum of soybean allergic patients and polyclonal antibody against soybean proteins. Soybean proteins were extracted by one-hour heating in boiling waterbath and separated by SDS-PAGE. After heat treatment, no significant changes of soy protein patterns were observed in SDS-PAGE analysis. Furthermore, the heat treatment had no effect on the results in immunoblotting with polyclonal antibody as well as in ELISA with soybean allergic patients' serum. With these results it may be concluded that allergenicity and antigenicity of soybeans do not reduce by thermal treatment.

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Allergenicity of Soybeans Depending on Their Variety (품종에 따른 콩의 알레르기성)

  • Son, Dae-Yeul;Kim, Ye-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.627-631
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    • 2010
  • The allergenicity of soybeans was analyzed using polyclonal antibodies and the blood sera of patients with soybean allergy, using fourteen different varieties of soybeans that are consumed in Korea. The study that used polyclonal antibodies having specificity for soybeans showed that while some of the fourteen varieties of soybeans contained additional protein bands indicating antigenicity, others lacked such bands, and the most antigenic protein was found in Jinpum soybeans. In comparing blood sera reactivity of four patients having soybean allergies, who had antigenicity values of 65U/ml or more according to CAP testing, the soybean varieties of Danbaek and Shinpaldal2 had the most reactivity and Daewon had the least. The result that the allergenicity of proteins in soybeans differs according to the variety of soybean, leads to the conclusion that it may be possible to reduce consumer allergic reactions to soybean products by choosing an appropriate variety of soybeans.

Property Comparison of Polyclonal Anti-Soy Protein Antibodies Produced for ELISA (효소면역측정법을 위한 다클론 항대두단백 항체의 생산 및 특성비교)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Yun, Sung-Seob
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1221-1226
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    • 2000
  • Specific antibodies were produced to develope the enzyme-linked immunosorbent assay for analysis of soy proteins and the properties of the antibodies were compared. Isolate soy protein(ISP), and ISP heated with SDS and urea (ISP(SU)), acidic subunits(AS) of 11S globulin were immunized to produce polyclonal antibodies. By using competitive indirect ELISA(ciELISA), the reactivities of the antibodies toward soy proteins treated with different methods were investigated and shown as $IC_{50}$. $IC_{50}'s$ of anti-ISP antibodies to ISP, ISP(SU), ISP treated with 2-ME(ISP(ME)), and crude 11S were 20, 30, 36, and $1000\;{\mu}g/mL$, respectively. And the values of anti-ISP(SU) antibodies to the same antigens were 100, 5, 4, and $220\;{\mu}g/mL$ and those of anti-AS antibodies were 20, 2, 2.5, and $200\;{\mu}g/mL$, respectively. Therefore, anti-AS antibodies showed the highest reactivities toward soy proteins among the produced antibodies as determined by ciELISA.

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Immunodiagnosis of clonorchiasis using a recombinant antigen (간흡충 재조합항원을 이용한 간흡충증의 면역 진단)

  • 용태순;양혜진
    • Parasites, Hosts and Diseases
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    • v.36 no.3
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    • pp.183-190
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    • 1998
  • A cDNA expression library of Clonorchis sinensis adult worm was constructed, and screened out immunologically. One clone, pBCs31, was selected ill view of its predominant reactivity with an experimentally infected rabbit serum. Recombinant C. slnensis antigen iIi 28 kDa as a if-falactosidase fusion protein produced in EscherichiG coli was identified by immunoblot analysis. The cloned gene was composed of 16 copies or a 30 base pair repeat and an additional 320 bases. The deduced amino acid seqiLence of the tandem repeat was AQPPKSGDGG. On RNA slot blot analysis, C. sinensis adult worm RNA showed a positive reaction with the cloned gene Enzyme-linked immunosorbent assay using a purified recombinant antigen of pBCs31 showed higtl specificity for diagnosis of clonorchiasis.

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