• Journal of Animal Science and Technology
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• 제46권3호
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• pp.495-502
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• 2004

장출혈성대장균 O157:H7의 eae유전자 산물인 intimin은 장 상피세포의 부착성을 조절하는 단백질로 알려져 있다. Intimin의 C-말단 아미노산 잔기와(His)$_6$이 혼합되어 이루어진 plasmid를 작성하여 재조합 단백질(34kDa)을 발현시켰다. 발현된 재조합 intimin 단백질의 면역원성을 확인하기 위해 토끼와 산란계에서 항혈청 및 난황항체(IgY)를 제조하였다. O157:H7에서 분리한 세포외막단백질(OMPs)과 제조된 항혈청과의 western blot상의 반응에서 약 94kDa의 위치에서 양성반응을 보여 발현된 재조합 단백질의 면역원성 효과가 확인되었다. 면역원성이 확인된 단백질을 산란계에 면역한 결과, 면역 후 4${\sim}$6주 경부터 높은 수준의 항체가 형성되었다. 재조합 intimin 단백질에 대한 난황 항체의 항원결합능력 조사를 위하여 중화반응시험을 수행한 결과, 재조합 intimin 단백질에 대한 난황 항체가 O157:H7과 강하게 결합하는 것으로 나타났다. 따라서 본 연구에서 발현된 재조합 intimin 단백질은 토끼와 산란계의 면역반응을 유도할 수 있는 효율적인 면역원임과 동시에 난황 항체 생산을 위한 항원으로의 이용가능성이 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.571-571
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• 2005

• Korean Chemical Engineering Research
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• 제50권5호
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• pp.866-873
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• 2012

난황에 포함된 IgY는 포유동물에 있는 바이러스나 항원에 반응하는 항체와 같은 역할을 한다. 난황을 전처리한 후 3-zone와 4-zone SMB를 이용하여 지질단백질들로 부터 IgY를 분리하는 전산모사연구를 수행하였다. 회분식 크로마토그래피에서 전산모사 매개변수와 흡착 등온식 SMB 전산모사 변수를 얻었다. Aspen simulator를 이용하여 전산모사를 수행하여 IgY를 분리할 수 있는 3-zone과 4-zone SMB 운전조건을 비교하여 다음과 같은 결과를 얻었다. IgY와 다른 단백질의 농도와 순도를 모두 고려할 때, 꼭지점인 좌표($m_2$, $m_3$=0.1, 1.1)에서 3-zone SMB가 최적의 조건으로 생각된다. IgY 만을 고려하면 4-zone SMB가 좌표($m_2$, $m_3$=0.06, 0.5)에서 가장 높게 IgY를 분리할 수 있었다. recycle이 없는 3-zone SMB는 꼭지점 좌표에서 좌표이동이 extract의 지질 단백질 농도에 큰 영향을 주었다.

• 한국식품과학회지
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• 제30권5호
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• pp.1029-1034
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• 1998

충치유발균인 Streptococcus mutans (혈청형 C)를 면역한 산란계의 난황에서 항체를 분리하고 그 특성을 조사하였다. 먼저 난황으로 부터 항체의 추출효율을 검토하였을 때, ${\lambda}-carrageenan$, $gammaYolk^{TM}$, $EGGstract^{TM}$의 방법으로 얻은 추출물 중 항체의 순도는 전기영동에서 각각 20, 46, 48%로 나타났으며, 난황 1 g에서 얻은 항체의 수율은 각각 11.3, 1.7, 1.8 mg이었다. 정량면역침강반응의 결과, ${\lambda}-carrageenan$방법으로 얻은 조난황 항체 중 특이항체의 비율은 12.2%이었다. 이로부터 계란 하나분인 15 g의 난황에서 얻은0.85 g의 조난황 항체에는 특이항체가 100 mg가량 함유되어 있음을 알 수 있었다. 세 종류 충치유발균주(혈청형 b, c, f)에 대한 특이항체의 반응성을 조사하였을 때, 설탕 첨가배지에서 배양한 균주는 C(+), f(+), b(+)의 순으로 48%이상의 높은 반응성을 나타내었으나 무설탕 배지에서 배양한 균주의 반응성은 각각 이보다 낮게 나타났다. 조난황 항체의 열 및 pH안정성은 양호하여, $70^{\circ}C$까지 50%이상의 활성을 유지하였고 $pH\;4{\sim}8$에서 대체로 안정하였다.

• 한국축산식품학회지
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• 제24권2호
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• pp.182-188
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• 2004

Lactoperoxidase(LPO)를 농도를 측정하기 위한 ELISA을 개발하기 위해 LPO로 면역시킨 갈색 산란계의 계란에서 형성된 anti-LPO IgY 항체를 분리 정제하고, 분리된 anti-LPO IgY 항체의 특이성을 ELISA 와 double immunodiffusion 방법으로 조사한 후 indirect ELISA 방법을 이용한 표준곡선을 만들었다. 분리 정제된 anti-LPO IgY항체의 titer는 1:520,000이며, ELISA와 double immunodiffusion 방법 모두에서 $\alpha$-lactalbumin, $\beta$-lactoglobulin, casein 및 lysozyme하고는 교차반응을 하지 않고 LPO만 높은 특이성을 갖는 항체로 나타났다. Indirect ELISA방법에서 LPO의 coating 농도는 0.25 $\mu\textrm{g}$/mL이며 anti-LPO IgY 최적 희석배수는 1:8,000으로 나타났다. Indirect ELISA 방법으로 LPO를 측정할 수 있는 표준곡선에서 민감도의 범위는 0.0l-l $\mu\textrm{g}$/mL로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.311-320
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• 2019

Fusobacterium nucleatum is a morbific agent in periodontitis and halitosis. Egg yolk antibody (IgY) was obtained from egg yolks from chickens stimulated with F. nucleatum. This study was to assess the effectiveness of IgY on periodontitis and halitosis caused by F. nucleatum in vitro and in vivo. The growth of F. nucleatum was inhibited (p <0.05) by different concentrations of IgY in vitro and the results of a Halimeter show volatile sulfur compounds (VSCs) were reduced to $904{\pm}57ppb$ at a concentration 40 mg/ml of IgY. The changes of fatty acids of F. nucleatum were determined using GC-MS. The scores for odor index of rat saliva were decreased. The major constituent of volatile organic compounds (VOCs) including short-chain acids decreased 46.2% in 10 mg/ml IgY, ammonia decreased 70% in 40 mg/ml IgY, while aldehydes and olefine ketones were almost unchanged. The ELISA assay revealed that IL-6 and TNF-${\alpha}$ were decreased after 4 weeks' IgY treatment. Morphometric (X-ray) and histological analyses (HE) showed that IgY reduced alveolar bone loss and collagen fibers became orderly in rat models. As a result, IgY may have the potential to treat periodontitis and halitosis.

• Animal cells and systems
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• 제5권1호
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• pp.85-90
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• 2001

Yolk platelets, one of the main food stores in the embryonic development, are composed of proteins. However, little is known about the identity of proteins utilized at certain stages of embryogenesis. In this study, we followed the fates of embryonic storage proteins by using an anti-polyubiquitin monoclonal antibody (mAB) as a probe. The mAb recognized the major storage proteins of Drosophila, Xenopus and chicken eggs. In the Drosophila embryo, the mAb-reactive 45-kDa protein was not used until stage 11 but was used up at stage 16 when the embryo completed segmentation. In the Xenopus embryo, the mAb-reactive 111 kDa protein was mostly utilized between stages 42 and 45 implying that the protein might be an energy source used just prior to feeding on food. By N-terminal sequencing the storage protein of Xenopus embryo was identified as a lipovitellin 1. This study confirms that storage proteins are used almost simultaneously at certain stages of embryogenesis and that vitellogenin 1 is the last storage protein in Xenopus embryogenesis.

• 한국미생물·생명공학회지
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• 제50권3호
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• pp.430-435
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• 2022

The infectious bursal disease (IBD) is a highly contagious and acute poultry disease caused by Birnavirus. However, the vaccination is the only disease prevention, but several factors impeded vaccine development. Thus, a need for time to develop a novel technique for managing and treating respiratory diseases in poultry birds. Passive immunization is a hope and a possible alternative used in birds to meet this need. The current research attempted to produce egg yolk-based polyclonal antibodies against the IBD virus. The benefits of IgY include ease of extraction, lack of reaction with mammalian Fc receptors, and low production cost. Commercial layers were immunized with inactivated IBD virus subcutaneously according to the treatment regimen. The eggs were gathered daily, and yolk antibodies were extracted with the ammonium sulfate precipitation technique. The use of an indirect hemagglutination test demonstrated that IgY was IBD-specific. Until the end of the experiment, the specific IgY immunoglobulins did not lose activity when stored at 4℃. The specific immunoglobulin (IgY) treated challenged birds were demonstrated 92% recovery in comparison to the control group. The study implies that the IBDV specific IgY is an easily prepared and rich source of antibodies and offers an alternative therapeutic agent to cure IBD-infected birds.

• Asian-Australasian Journal of Animal Sciences
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• 제17권8호
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• pp.1139-1144
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• 2004

In Exp. 1, a total of 36 pigs (6.55$\pm$0.10 kg average initial body weight and 21 d average age) were used in a 14 d growth study to determine the effects of replacing spray-dried plasma protein (SDPP) with spray-dried egg protein containing specific egg yolk antibody (SDEP) on growth performance and nutrient digestibility in weaned pigs. The pigs were blocked by weight and assigned to treatments based on sex. There were three pigs per pen and four pens per treatment. Dietary treatments were 0, 3, or 6% SDEP and contained 6, 3, or 0% SDPP, respectively. Through the entire experimental period, average daily gain (ADG), average daily feed intake (ADFI), and gain/feed tended to decrease as the concentration of SDEP increased in the diets. However, there were no significant differences among the treatments (p>0.05). As the addition of SDEP in the diets increased, apparent digestibilities of dry matter (DM) and nitrogen (N) were decreased without significant (p>0.05). For Exp. 2, 36 pigs (2.63$\pm$0.04 kg average initial body weight and 10 d average age) were used in a 14 d growth study to determine the effects of antibiotic replacement with SDEP on growth performance and nutrient digestibility in early-weaned pigs. The pigs were blocked by weight and assigned to treatments based on sex. There were three pigs per pen and four pens per treatment. Dietary treatments included 1) ANTIBIOTIC (corn-dried whey-soybean meal based diet+0.08% antibiotics, 4 mg of tiamuline hydrogen fumarate; 10 mg of sulfadimidine per kg of complete diet), 2) SDEP0.1 (corndried whey-SBM based diet+0.1% SDEP), and 3) SDEP0.2 (corn-dried whey-SBM based diet+0.2% SDEP). ADG and gain/feed of pigs fed the SDEP0.2 diet were higher than for pigs fed the ANTIBIOTIC diet without significant (p>0.05). Pigs fed the diet with SDEP0.2 tended to have increased apparent digestibilities of DM and N compared to pigs fed the ANTIBIOTIC diet without significant (p>0.05). In conclusion, the dietary SDEP seemed to be partial replacing the SDPP portion of high nutrient dense diet for weaned pigs. Also, dietary SDEP seemed to be approximately 0.2% or more when the pigs fed the antibiotic-free diet for early-weaned pigs.