• Title/Summary/Keyword: 다클론 항체

Search Result 30, Processing Time 0.043 seconds

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
    • /
    • v.49 no.3
    • /
    • pp.196-201
    • /
    • 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
    • /
    • v.36 no.3
    • /
    • pp.369-374
    • /
    • 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.

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
    • /
    • v.41 no.1
    • /
    • pp.49-53
    • /
    • 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.

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
    • /
    • v.32 no.6
    • /
    • pp.1221-1226
    • /
    • 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.

  • PDF

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
    • /
    • v.46 no.5
    • /
    • pp.47-55
    • /
    • 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.

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
    • /
    • v.9 no.2
    • /
    • pp.135-142
    • /
    • 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).

  • PDF

Effects of Polyclonal Antiserum Against Adipocyte Plasma Membrane Proteins on Body Composition of Passively Immunized Sprague-Dawley Male Rats (지방세포 원형질막 단백질에 대한 다클론 항체의 수동면역이 수컷 흰쥐의 체조성에 미치는 영향)

  • Baek, K.H.;Choi, C.B.
    • Journal of Animal Science and Technology
    • /
    • v.44 no.1
    • /
    • pp.39-44
    • /
    • 2002
  • The current study was conducted to investigate the effects of administration of antiserum against adipocyte plasma membrane(APM) proteins into rats on body fat mass. Twenty(20) male adult Sprague-Dawley rats were randomly allocated into either control or antiserum treatment group(10 rats/treatment) and immunized with physiological saline(control group) and polyclonal antiserum (treatment group), respectively, raised in sheep against rat APM proteins(5times, 2day interval). All animals were killed 4weeks after last injection. Intraperitoneal(i.p.) administration of antiserum significantly(P=0.0054 and P=0.0019, respectively) reduced subcutaneous(21.9%) and perirenal + mesentric + epididymic(36.0%) adipose tissue mass in rats of treatment group. Although body weights of antiserum treated rats were decreased during immunization, the rats recovered their body weight after 1 week of treatment. There were no significant changes in the level of blood glucose and in the contents of muscle protein and fat in antiserum treated animals. Current results indicate that polyclonal antibodies against APM proteins could be used to manipulate body fat mass in meat animals as well as laboratory animals. Further studies, however, are necessary for the practical applications of the current results.

Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocytes for Fat-Reduced Hanwoo Beef Production (한우 체지방 감소 쇠고기 생산을 위한 복강 및 피하지방 항체 개발)

  • Choi, Chang-Weon;Kim, Yu-Hyun;Kim, Sang-Jin;Song, Man-Kang;Kwon, Eung-Gi;Oh, Young-Kyoon;Hong, Seong-Koo;Choi, Seong-Ho;Baek, Kyung-Hoon
    • Food Science of Animal Resources
    • /
    • v.28 no.5
    • /
    • pp.651-659
    • /
    • 2008
  • This study aimed to develop polyclonal antibodies to regional inedible adipocytes of Korean native cattle (Hanwoo) and investigate cross-reactivity of the antibodies. Patterns in plasma membrane proteins (PMPs) from abdominal and subcutaneous adipocytes of Hanwoo isolated by collagenase digestion were investigated using SDS-PAGE. As antigens, abdominal and subcutaneous adipocyte PMPs of Hanwoo were injected to sheep 3 times at 3 wk intervals for passive immunization, and non-immunized serum and antisera were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with heart, kidney, liver, lung, muscle, and spleen of Hanwoo were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes of Hanwoo were performed for analysing LDH concentration. Based on the SDS-PAGE analysis, specific proteins of PMPs in abdominal and subcutaneous adipocytes appeared despite rather similar patterns between both adipocytes. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas the antisera had relatively strong reactivity up to the level of 1:128,000 and 1:64,000 dilution. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivities of abdominal and subcutaneous adipocyte antisera were detected with PMPs of the organs. Both antisera strongly reacted with each adipocyte PMPs and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and have safety in cross-reactivities with body organs. Further studies on in vivo cross-reactivity and fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo should be required for inedible fat-reduced high quality beef production.

Development of Enzyme-Linked Immunosorbent Assay for Glyphosate-Tolerant Soybeans (제초제내성 유전자재조합 콩의 검출을 위한 면역분석법 개발)

  • Kwak, Bo-Yeon;Ko, Seung-Hee;Park, Chun-Wuk;Son, Dae-Yeul;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
    • /
    • v.35 no.3
    • /
    • pp.366-372
    • /
    • 2003
  • Enzyme-linked immunosorbent assay (ELISA) for assaying the 5-enolpyruvyshikimate-3-phosphate synthase from Agribacterium sp. CP4 (CP4 EPSPS) in genetically modified soybeans was developed. Polyclonal and monoclonal antibodies (Pab, Mab) specific to the CP4 EPSPS were produced. When using the Pab, the detection limit of sandwich ELISA toward CP4 EPSPS (0.03 ${\mu}g/mL$) was better than that of competitive indirect ELISA(ciELISA) (1 ${\mu}g/mL$). It was found that 2 of 3 monoclonal antibodies, Mab1 and Mab2, recognized the same antigenic determinant on CP4 EPSPS, but Mab3 recognized different antigenic determinant when competitive ELISA was performed using the Mabs. On the other hand, when the sensitivity of sandwich ELISA using combination of Pab and/or Mabs was determined, the sandiwich ELISA using Mab2 as a capture antibody and Pab-HRP as a secondary antibody showed the lowest detection limit of CP4 EPSPS (0.02 ${\mu}g/mL$). The sandwich ELISA developed in this study could be applied to detect glyphosate-tolerant soybeans.

Effect of Serum Type on Hybridoma Growth and Monoclonal Antibody Production (하이브리도마 세포증식과 단일클론항체 생산에 미치는 혈청 종류의 영향)

  • 전복환;박송용
    • KSBB Journal
    • /
    • v.9 no.3
    • /
    • pp.253-265
    • /
    • 1994
  • We have studied the effects of serum concentration and initial cell density on hybridoma cell growth and monoclonal antibody (MAb) production at various media supplemented with different types of serum. The types of serum were fetal bovine sera, newborn bovine calf sera, calf sera including supplemented calf sera, horse serum, and goat serum. The concentrations of each serum were 0.5, 1.25, 2.5, and 5% (v/v) and the inoculum densities were $5{\times}10^4, 1{\times}10^5, 2{\times}10^5,$ cells/ml. The hybridoma cell growth and anti-Hepatitis B surface antigen (anti-HBsAg) MAb production were found to be enhanced by increasing the serum concentration and by increasing inoculum density regardless of serum type. We found that test sera purchased from different companies show different effects on cell growth and MAb production, although they are the same type of serum.

  • PDF