• Title/Summary/Keyword: yolk antibody

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Production of a Specific Yolk Antibody against Enterotoxigenic E. coli F41 Fimbrial Antigen (장독성대장균 F41 섬모항원에 대한 특이난황항체 생산)

  • Shin, S.O.;Kim, J.W.
    • Journal of Animal Science and Technology
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    • v.44 no.5
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    • pp.633-642
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    • 2002
  • Enteric colibacillosis has economically become an important disease of young animals as a result of increasing intensification of farrowing management. The objective of this experiment is to isolate fimbrial antigen from enterotoxigenic E. coli F41, to develop specific polyclonal IgY which can effectively neutralize or reduce the proliferation of pathogens in feed or living animal system, and to apply IgY technologies to animal industry. The results obtained were as follows: The molecular weight of the purified F41 antigen was 29,500 dalton on sodium dodecyl sulfate-polyacrylamide gels. Fimbrial antigen was confirmed by the western blot method. It was observed that after immunization the level of serum antibody titer of laying hen was shown in two weeks and gradually increased. The antibody titer in egg yolk appeared two weeks after it was shown in serum antibody. The titers of egg yolk antibody were gradually increased to the maximum level of 320,000 (antigen 50${\mu}g$/$m\ell$), 450,000 (antigen 200${\mu}g$/$m\ell$), and 320,000 (antigen 600${\mu}g$/$m\ell$). According to the results of specificity test by ELISA, the anti-F41 antibodies from chicken serum and egg yolk reacted only with ETEC F41 antigen. There was no cross reaction with other ETEC strains (K88, K99, and 987P). In vitro condition, as a result of antigen binding ability of yolk antibodies, bacterial concentration was rapidly decreased to $10^5$ CFU/$m\ell$ from $10^9$ CFU/$m\ell$ when 2${\sim}$4 mg/$m\ell$ of freeze dried WSF (water soluble fraction) was added.

Effects of Egg Yolk Antibodies Produced in Response to Different Antigenic Fractions of E. coli O157:H7 on E. coli Suppression

  • Chae, H.S.;Singh, N.K.;Ahn, C.N.;Yoo, Y.M.;Jeong, S.G.;Ham, J.S.;Kim, D.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.11
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    • pp.1665-1670
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    • 2006
  • The objective of this research was to provide the characterization and method for producing anti-E. coli O157:H7 antibodies in egg-laying hens and to determine if the antibody can restrain the proliferation of E. coli O157:H7 in-vitro. Selected antigenic fractions (whole cell, outer membrane protein and lipopolysaccharide (LPS)) from E. coli O157:H7 were injected to hens in order to produce anti-E. coli O157:H7 antibodies. The immune response and the egg yolk antibodies of laying hens against the whole cell, outer membrane protein and LPS antigens were monitored by ELISA. The level of antibodies against whole cell antigen monitored through ELISA sharply increased after the initial immunization, and it was found to be maximum on day 49 however, the level was maintained up to day 70. Antibodies (5 mg/ml) directed against the whole cell inhibited E. coli proliferation 10-13 times more than outer membrane protein or LPS. The antibody response against the whole cell antigens appeared to have higher activity in restraining the proliferation of E. coli O157:H7 than antibody against outer membrane protein or LPS. Results reflected that increasing the IgY's in the egg yolk could prevent greater economic losses due to human and animal health from pathogenic bacteria i.e. E. coli O157:H7.

Immuno-specificity of Egg Yolk Antibodies against Bovine Rotavirus and Bovine Coronavirus causing Calf Diarrhea (송아지 설사병 주요원인체인 소로타바이러스와 소코로나바이러스에 대한 난황항체 생산 및 면역특이성 분석)

  • Lee, Seong;Woo, Seung-Eun;Lee, Sang-Rae;Kim, Jung-Woo
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.407-412
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    • 2009
  • This study was performed to produce specific egg yolk antibodies against bovine rotavirus (BRV) and bovine coronavirus (BCV) that are major pathogens causing diarrhea in calves. Chickens were immunized with BRV and BCV intramuscularly in the breast muscle by injection 5 times at two weeks interval. At 6 weeks after the first immunization of BRV or BCV, cross reactivity of each serum derived from BRV- or BCV-immunized hens was tested. Each serum antibody against BRV or BCV was reacted with only specific BRV or BCV antigens. Serum and egg yolk-antibody titers of hens against BRV or BCVwere highest at 8~12 weeks after first immunization. Specific serum and egg yolk-antibody titers against BRV were about 104,000 and 107,000, respectively, and those against BCV were about 145,000 and 155,000, respectively. Hemagglutination inhibition titers in the immunized egg yolk antibodies against BRV and BCV were 5,120 and 1,280, respectively, and were ${\geq}8$ times higher than that in non-immunized control. These results suggested that the immunized egg yolk antibodies could effectively neutralize BRV and BCV.

Development of preventive method for enterotoxigenic colibacillosis using egg yolk antibodies II. Therapeutic effect of egg yolk antibodies against colibacillosis of piglets (난황항체를 이용한 돼지 대장균설사증 방제기법 개발 II. 난황항체의 돼지 대장균증에 대한 치료효과)

  • Kim, Jong-man;Woo, Seng-ryong;Kweon, Chang-hee;Kim, Jong-yeom;Huh, Won
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.837-842
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    • 1998
  • The present study describes the effectiveness of egg yolk antibodies (IgY) against enteric colibacillosis and edema disease in piglets. The antibodies were gained from the egg yolk of hens immunized with k88, k99, 987p fimbrial adhesin and heat-labile toxin antigens of enterotoxigenic Escherichia coli (ETEC). Orally-administered egg yolk antibodies solution protected against experimental challenge with ETEC $K88^+$ and $k99^+$ strains in neonatal piglets and mice. In field trial, a total of 598 diarrheal piglets were orally treated with 3ml of antibody once a day to determine for the therapeutic effect. Of them, 582 (97.3%) piglets were recovered from diarrhea in 3 days. We also experimentally treated with the egg yolk antibodies twice a day for 5 consecutive days for 94 weaning piglets with edema disease for the determination of therapeutic effects. Seventy four piglets (78.7%) were recovered from clinical edema signs. Theses findings indicate that egg yolk antibodies against k88, k99, 987p and LT of ETEC are useful source of passive immunity for enteric colibacillosis and edema disease of piglets.

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Studies on the Stability of Hen′s Egg Yolk Immunoglobulins (난황 항체의 안정성에 관한 연구)

  • 이경애
    • Korean journal of food and cookery science
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    • v.12 no.1
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    • pp.54-59
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    • 1996
  • Immunoglobulins (IgY) were isolated from egg yolk of hens immunized with bovine serum albumin(BSA). The stability of anti-BSA IgY against heat and pH was investigated. Antibody activity was measured by enzyme linked immunosorbent assay. IgY was relatively heat-stable and most of the antibody activity remained after heating up 65$^{\circ}C$ for 30 minutes. IgY was stable at pH 5-11. However, inactivation of IgY was observed below pH 4, or above pH 12. Inactivation of IgY proceeded rapidly at low pHs(pH 2-3). Most of the antigen binding activity was lost at low pHs probably because of some conformational changes.

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Control of swine respiratory disease using egg yolk antibodies I. Analysis of immunogenes of Bordetella bronchiseptica, Pasteurella multocida and Actinobacillus pleuropneumoniae and production of IgY (난황항체를 이용한 돼지 호흡기 질병 방제 관한 연구 I. Bordetella bronchiseptica, Pasteurella multocida 및 Actinobacillus pleuropneumoniae의 주요 면역원 분석 및 IgY의 생산)

  • Shin, Na-ri;Kim, Jong-man;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.40 no.3
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    • pp.551-561
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    • 2000
  • Swine respiratory diseases have induced severe economic losses in swine industry worldwide. Several methods have been developed and applied to prevent and control the disease. However, those are still problematic in swine industry. Recently, the use of egg yolk antibodies with several advantages was introduced and applied to control diseases in animal as well as human. As the first step of the use of egg yolk antibodies in the control of the swine respiratory diseases, we investigated the immunogens of the causative agensts of the diseases and immune response in egg yolk of hens immunized with them. Bacterial antigens prepared from Bordetella bronchiseptica, Pasteurella multocida 3A and 4D, and Actinobacillus pleuropneumaniae serotype 2 and 5 were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot and toxicity test in mice. The antigens were injected into laying hens in order to produce antibodies against them in egg yolk. After chickens were immunized three times in 2 weeks interval, the profile of antibody production was examined by ELISA. The production of antibody in egg yolk was started in 2 weeks after the first injection, reached peak in 6-8 weeks and maintained until 12 weeks. Of two adjuvants used in this study, ISA70 was more effective than aluminum hydroxide gel in enhancing immunogenecity, laying rates and safety in hens. These results suggested that egg yolk antibodies could be a good source for production of antibodies specific to pathogenic bacteria inducing respiratory diseases of swine.

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Development of an anti-obesity dietary supplement inhibiting the digestion of carbohydrate and lipid (비만 개선 효과를 지닌 탄수화물 및 지방 흡수 억제 기능성 식이조성물 개발)

  • 윤유식;최선미;홍순복;홍정미;김정원;이홍석;홍성길
    • Korean journal of food and cookery science
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    • v.18 no.3
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    • pp.319-324
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    • 2002
  • In a previous study, we developed a new food additive as an egg yolk antibody (IgY) against carbohydrate digestion enzymer for the regulation of blood glucose level and weight control. The IgY delayed and decreased the increment if blood glucose level after administration of sucrose in human being by 30% in 20∼30 min. We also developed a lipase inhibitor as a water extract of two kinds of herb, Platycodon grandiflorum and Solanum Melongena, Twenty three volunteers were subjected to the intake of the egg yolk IgY Plus the herbal extracts for 50 days. In average, the treated subjects appeared to lose 1.96 kg of body weight and 3.4 kg of body fat mass during the treated period. Furthermore, Panniculus adiposus and breech size were significantly decreased during the experimental period. Above results suggested that the administration of the dietary additives composed of egg yolk IgY and natural herbal extract improve the obesity by the decrement of body weight and body fat mass.

An Experimental Study on Production of Egg Yolk Antibody(IgY) against Bee Venom (봉독의 항독소(IgY)생산을 위한 실험적 연구)

  • Hwang, Tae-Jun;Lee, Seung-Bae;Gwon, Gi-Rok
    • Journal of Pharmacopuncture
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    • v.4 no.2
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    • pp.5-15
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    • 2001
  • This study was carried out for production of neutral antibody to bee venom $(anti-phospholipase\;A_2IgY)$. Hen layings were injected repeatedly with bee venom and phospholipase $A_2$ with Freund's adjuvant. Specific antibody in egg yolk from immunized hen laying was separated, and purified, also immunological characteristics of anti phospholipase $A_2\;IgY$ was invested. The results were summarized as follows: 1. Phospholipase $A_2$ was showed single band at molecular weight 17,000 in SDS-PAGE and bee venom was showed two band at molecular weight 17,000 and under molecular weight 6,500 in SDS-PAGE. 2. During 70 days after hen immunized with bee venom and phospholipase $A_2$, antibodies(anti-bee venom IgY) to bee venom were showed poor ELISA value in egg yolk, but antibodies$(anti-Phospholipase\;A_2IgY)$ to phospholipase $A_2$ in egg yolk were increased ELISA value from 8 days or 15 days and found maximum ELISA value at 42 days. Also after booster at 49 days, ELISA value of anti Phospholipase $A_2\;IgY$ in egg yolk was supported at optical density(O.D) 1.0 level, continuously. 3. Titer of phospholipase $A_2\;IgY$ was showed 1: 32,000. 4. In double immunodiffusion test to phospholipase $A_2$ after double dilution of anti-phospholipase $A_2\;IgY$, only precipitation line was made in 1:1 dilution well of anti-Phospholipase $A_2\;IgY$. But In immunodiffusion test to anti-phospholipase $A_2\;IgY$ after double dilution of phospholipase $A_2$, Precipitation line to 250ul/ml well of phospholipase $A_2$ was showed. In double immunodiffusion test to bee venom(1mg/ml) after double dilution anti-phospholipase $A_2\;IgY$, all well without 1:32 dilution well were showed strong precipitation line. 5. In dot bloting test to anti-phospholipase $A_2\;IgY$ after diluting bee venom(0.5mg/ml), dot bloting color was showed clearly to $1/100(5{\mu}g/ml)$ in bee venom.

The Production of Egg Yolk Immurnoglobulin (IgY) Raised against 3T3L-1 Cell Membrane Protein and the Control of Adipocytes Differentiation (3T3L-1세포의 막단백질에 대한 난황면역글로뷸린 (IgY)의 생산과 지방세포의 분화조절작용)

  • 김상윤;황성구;구의섭;고태송
    • Korean Journal of Poultry Science
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    • v.26 no.3
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    • pp.179-188
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    • 1999
  • The present was undertaken to establish a model for the control of adipocytes differentiation by using antibody from egg yolk. The emulsion of membrane protein of 3T3L-1 cell membrane protein with the complete Freund's adjuvant was firstly immunized in layer. Second and third boosting were undertaken with two weeks intervals by injection of the emulsion of the same antigen with the incomplete Freund's adjuvant. After 4 week of the first immunization, eggs were collected and antibody (IgY) was purified from egg yolk. The purity of IgY was 60-98% determined by single radial immunodiffusion (SRID) methods. Titer value of the antibody showed high reactiviy for the preadipocytes membrane protein measured by ELISA. When the IgY was added in the test media containing either 2.5% porcine serum or 10% FBS(control), the differentiation of 3T3L-1 cells and Glycerol-3-phosphate dehydrogenase(GPDH) activities was significantly decreased compared to the control cells(p〈0.05). When mice were subcutaneously injected with IgY raised against membrane protein of 3T3L-1 cells for 3 weeks, adipose tissue mass around ovary was tended to be decreased in female mice compared to those of control mice. It is suggested that a potential for manipulating of lipid accumulation through decrease in 3T3L-1 cell differentiation and fat accumulation in female mice by IgY treatment.

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Production of Immunospecific Egg Yolk Antibody with Recombinant Staphylococcal Enterotoxin B (SEB) Protein (포도상구균에서 분비하는 장내독소 B(SEB)에 대한 재조합 단백질을 이용한 면역특이적 난황항체 생산)

  • Lee, Seong;Lee, Sang-Rae;Jung, Kyung Min;Kim, Jung Woo
    • Korean Journal of Poultry Science
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    • v.39 no.4
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    • pp.273-278
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    • 2012
  • Staphylococcal enterotoxin B (SEB), which is a bacterial superantigen produced by Staphylococcus aureus, is associated with serious diseases, including food poisoning and atopic dermatitis. This study was performed to produce about 30 kDa of recombinant SEB protein and to immunize in chickens to acquire the specific egg yolk antibody (IgY) against the recombinant SEB. Chickens were immunized with the recombinant SEB intramuscularly in the breast muscle by injection 3 times at intervals of two weeks. Serum- and egg yolk-antibody titers of hens against SEB were highest at 4 weeks after first immunization. In western blot, anti-recombinant SEB IgY was reacted immunospecifically against the recombinant SEB and commercialized SEB. These results suggested that the recombinant SEB antigen could be used as an immunogen to elicit antibody (IgY) against SEB and the anti-recombinant SEB IgY could neutralized staphylococcal enterotoxin B effectively.