• BMB Reports
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• 제28권1호
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• pp.27-32
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• 1995

Polyclonal antibodies against human DNA-dependent RNA polymerase II (HPP II) were generated from chicken egg yolk after immunization with RNA polymerase II as an antigen. The antibodies from egg yolk (IgY) were purified and characterized. IgY showed a specificity against DNA-dependent RNA polymerase II, and was a polyclonal antibody against 12 subunits of polymerase II. An amount of 0.35 mg of IgY was obtained freman HPP II-Sepharose affinity column using 10 eggs from a chicken immunized against RNA polymerase II as an antigen. These antibodies can be used for isolating the genes for RNA polymerase II components, and for in vitro transcription assays using HP-RNA polymerase II.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.79-83
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• 2000

A han transfers her serum immunoglobulin G to the agg (IgY) and gives immunity to her offspring. Therefore, The hen agg can be an effective supplier of a large amount of antigen specific antibody that accumulates in the egg yolk. Antigen specific antibody has been widely used for immunological analysis in the field of diagnosis as well as pure scientific research. The production and separation technology of IgY is demonstrated in the present study.

• 한국가금학회지
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• 제25권4호
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• pp.161-167
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• 1998

가금 및 가축의 살모넬라중을 일으키는 에서 편모항원을 순수분리하고 이를 면역원으로 산란계에 면역하여 난황 중에 형성된 항체와 혈청간에 면역 반응을 조사하고 생산된 에 대한 특이난황 항체의 특이성 검사와 항체회수율 검사를 실시한 결과S. enteritidis는 다음과 같다. 1. S. enteritidis에서 순수 분리한 flagella protein 의 분자량은 54.6 kDa으로 나타났다. 2. 산란계의 혈청중 항체가 형성은 면역 후 2주경부터 급격히 증가하기 시작하여 6주경에 최고의 항체가 수준에 도달하였다. 반면에 난황중 항체가는 면역 후 4주경부터 급격히 증가하기 시작하여 6주경에 혈청항체가 수준과 동일하게 도달되며, 10주 이후에는 혈청항체가 수준보다 높게 형성되어 일정한 수준을 유지하였다. 3. S. enteritidis의 flagella protein을 면역원으로 산란계에 면역하여 생산된 난황항체는 주로 S. enteritidis 균주에서만 특이적으로 반응하며 E. coli (Kl2:K99, K88+, 987P) 균체에서는 반응이 전혀 없는 것으로 나타났다. 4. 한개의 난으로부터 분리한 IgY의 함량은 106 mg이었으며 회수율은 88.3％였다.

• 한국식품조리과학회지
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• 제18권1호
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• pp.94-100
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• 2002

The dietary carbohydrates are mainly digested and adsorbed at small intestine. We developed a new food additive as an egg yolk antibody(1gY) against maltase, sucrase and sodium dependent g1ucose cotransporter(SGLT) for the regulation of blood glucose level and weight control. The maltase, sucrase and SGLT were purified from porcine small intestine which is very similar to that of human in physiological characteristics. The purification step contained an ultracentrifugation, ion exchange chromatography and hydrophobic chromatography. The hens were immunized by purified protein and the IgY activities against immunized antigens were determined. This antibody obtained from the immunized hen's egg yolks directly inhibited the activities of maltase and sucrase in vitro. And the IgY delayed and decreased the increment of blood g1ucose level after administration of maltose, sucrose and glucose in rat about 30 to 60%. The results of this study suggest that the IgY inhibiting the carbohydrate digestion could be used as functional food materials for weight control and regulation of blood glucose level in diabetes.

• 한국식품조리과학회지
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• 제12권1호
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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.

• 한국식품위생안전성학회지
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• 제18권2호
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• pp.61-66
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• 2003

본 연구는, Vibrio parahaemolyticus에 대한 면역 글로블린 Y(IgY)를 생산하기 위하여 시행되었다. Vibrio parahaemolyticus는 중요한 병원성 균이며, 위장염을 유발시키는 원인 균이다. 따라서, 난황에서 Vibrio parahaemolyticus에 대한 항체를 생산하기 위하여 항원 LPS(lipopolysaccharide)를 암탉에 주사하였다. 면역은 2주 간격으로 4차까지 실시하였으며, 추가면역으로 2주 간격으로 3차까지 실시한 후 ELISA로 그 항체 역가를 측정하였다. 난황에서의 항체생성은 1차 면역 후 1주일부터 나타나기 시작하였으며, 7주에 최고 역가에 도달하였고, 이러한 역가는 13주 이후까지 지속되는 것으로 나타났다. 한편 혈청에서의 항체 역가도 난황에서의 경향과 유사하였다. 정제시킨 IgY를 본 연구의 water-dilution방법과 상업용 IgY분리 kit로 그 정제 정도를 비교해본 격과 큰 차이를 나타내지 않았으며, Vibrio parahaemolyticus외의 다른 Vibrio 속 및 세균들과의 교차반응성 조사에서 특별한 교차반응이 없었다. 본 연구에 나타난 결과로, 아주 적은 양의 LPS 항원으로 역가가 높은 항체의 생산이 가능하였으며, 이러한 결과들로 보아 난황은 유행성 위장염을 유발하는 병원성 균의 특이 항체 개발을 위한 좋은 공급원이 될 것으로 생각된다.

• 대한약침학회지
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• 제4권2호
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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.

• 대한수의학회지
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• 제42권3호
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• pp.371-376
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• 2002

Swine respiratory diseases have induced severe economic lasses in swine industry worldwide. Therefore, several methods have been made and applied to prevent and control the diseases. However, these methods still have a problem and also induce side effects. Recently, the use of egg yolk antibody was introduced to control and prevent the diseases as one of new trials. As a study of using egg yolk antibody, antibody titers against several different antigens of major pathogens in swine respiratory diseases were compared in egg yolk and serum of hens immunized with those antigens. The titers were measured by ELISA using the antigens as coating antigens. The relationship in antibody titers between egg yolk and serum were identified by analysis of variance for linear regression. Almost of antigens used in this study showed the high relationship in antibody titers between egg yolk and serum (r = 0.87 ~ 0.93) even though the relationship in antibody titers against P. multocida A:3 IROMP was slightly low (r = 0.74)(P<0.01). These results indicated that antibody titer in egg yolk could be useful to predict the titer in serum of chicken.

• 한국가금학회지
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• 제31권1호
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• pp.37-44
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• 2004

It has been recognized that the hen, like its mammalian counterparts, provides young chicks with antibodies as protection against hostile invaders. This system facilitates the transfer of specific antibodies from serum to egg yolk, and provides a supply of antibodies called immunoglobulin Y(IgY) to the developing embryo and the hatched chick. The protection against pathogens that the relatively immune-incompetent newly hatched chick has, is through transmission of antibodies from the mother via the egg. Egg yolk, therefore, can be loaded with a large amount of IgY against pathogens which can immobilize the existing or invading pathogens during the embryo development or in day-old chicks. Thus, the immunization of laying hens to various pathogens results in production of different antigen-specific IgY in eggs. Egg yolk contains 8∼20 mg of jmmunoglobulins (IgY) per ml or 136∼340 mg per yolk suggesting that more than 30 g of IgY can be obtained from one immunized hen in a year. By immunizing laying hens with antigens and collecting IgY from egg yolk, low cost antibodies at less than $10 per g compared to more than $20,000 per g of mammalian IgG can be obtained. This IgY technology opens new potential market applications in medicine, public health, veterinary medicine and food safety. A broader use of IgY technology could be applied as biological or diagnostic tool, nutraceutical or functional food development, oral-supplementation for prophylaxis, and as pathogen-specific antimicrobial agents for infectious disease control. This paper has emphasized that when IgY-loaded chicken eggs are produced and consumed, the specific antibody binds, immobilizes and consequently reduces or inhibits the growth or colony forming abilities of microbial pathogens. This concept could serve as an alternative agent to replace the use of antibiotics, since today, more and more antibiotics are less effective in the treatment of infections, due to the emergence of drug-resistant bacteria.

• 동아시아식생활학회지
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• 제9권2호
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• pp.200-206
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• 1999

Salmonella typhimurium 항원에 면역된 산란계의 계란항체(IgY)생산 및 증가를 측정한 결과, 산란계가 이들 항원에 면역반응을 일으켜 2주 후 혈청과 난황에서 정상군에 비하여 뚜렷한 항체가 생성능을 보였으며. 혈청내의 IgG생성에 비하여 계란항체(IgY)의 생성이 다소 낮은 것으로 나타났다. 혈청 및 난황내의 단백질 함량은 정상군에 비하여 약간 높은 경향을 나타내어 산란계의 혈청 및 난황내의 단백질이 이들 항원에 의하여 증가하는 것으로 나타났다. DEAE-Sephacel column을 통과한 분획은 두 개의 peak를 나타내었다. 각 분획의 IgY농도를 ELISA법으로 확인한 결과 시험관 350-635$m\ell$에 해당되는 두번째 peak에서 대부분 측정되었다. 분리 정제된 IgY의 분자량을 측정한 결과, heavy chain 72-75KD, light chain 30-40KD정도였다. 이상의 결과에서, 본 실험에서 사용된 IgY는 항원에 대한 특이성 및 생화학적 특성 등을 고려해 볼 때, 식품산업 소재, 의약품 소재, 연구용 kit와 진단용 kit 및 축산분야 등 여러 분야에 응용될 수 있으리라 기대된다.