• 약학회지
• /
• 제49권1호
• /
• pp.6-10
• /
• 2005

Immunoglobulin Y (IgY) obtained from chicken as the immunization host brings several advantages to antibody production, such as improved yield, lower cost, longer stability and higher specificity than mammalian immunoglobulin. In the present study, we attempted to produce IgY against a sialic acid-binding lectin, Maackia fauriei agglutinin (MFA), from egg yolk of white Leghorn hens. For the isolation of IgY from egg yolk, we applied a water dilution method. The weekly yield of IgY was determined by enzyme-linked immunosorbent assay, with a final yield of anti-MFA IgY from total IgY of approximately $1\%$. The yielded IgY were used to prepare IgY-affinity column conjugated with CNBr-activated Sepharose 4B, which resulted in the lectin being successfully purified in a single step from Maackia fauriei. This purified lectin exhibited the same hemagglutination activity as lectin purified using conventional purification methods.

• 대한수의학회지
• /
• 제41권2호
• /
• pp.197-202
• /
• 2001

Although several methods have been developed and applied to control swine respiratory diseases, the disease induces severe economic impact to swine industry worldwide. As one of the new trials, application of egg yolk antibody(IgY) was attempted for the purposes and immune response in sera and egg yolk was analysed with ELISA in previous study. In this study, immunological specificity of the IgY was analysed by Western blot analysis. In the analysis of causative agents of atrophic rhinits, B bronchiseptica and P multocida 4D, proteins of 33, 40, 43, 67 and 141 kDa were specifically reacted with IgY Also, 40 and 110 kDa proteins were identified as the major immunogens in P multocida 3A. In A pleuropneumoniae serotypes 2 and 5, 40 kDa and 47 kDa proteins were found to be the major reactive ones. These results suggested that egg yolk antibodies from immunized hens was specific with antigens injected into hens and partially purified antigens, outer membrane proteins and dermonecrotic toxin, were more effective than bacterin for the production of specific antibody.

• 대한수의학회지
• /
• 제44권1호
• /
• pp.65-71
• /
• 2004

This study was carried out to produce IgY against B. bronchiseptica, parvovirus and distemper virus that are major pathogens in alimentary and/or respiratory diseases of dogs. In the comparison of adjuvants, ISA70 was the best in the rapid induction and maintence of antibody titers. Agglutination antibody titers against B. bronchiseptica were 1:1,280 ~ 1:10,240 in sera and 1:160 ~ 1:1,280 in egg yolk. Hemagglutination inhibition(HI) titers against parvovirus in sera and egg yolk were 1:80 ~ 1:320 and 1:64 ~ 1:256, respectively. Virus neutralization titers against canine distemper was 1:8 ~ 1:64 in sera and egg yolk. These results suggested that egg yolk antibody titers could be variable according to a sort of adjuvant and antigens of the pathogens.

• Korean Chemical Engineering Research
• /
• 제49권6호
• /
• pp.798-803
• /
• 2011

난황에 포함된 IgY는 포유동물에 있는 바이러스나 항원에 반응하는 항체와 같은 역할을 한다. IgY 분리를 위해 난황을 전처리한 후 3-zone SMB를 이용하여 지질단백질들로 부터 분리하는 전산모사연구를 수행하였다. 회분식 크로마토그래피와 pulse input method(PIM) 실험을 이용하여 전산모사 매개변수와 흡착 등온식을 얻었다. Aspen simulator를 이용하여 전산모사를 수행하여 IgY를 분리할 수 있는 SMB 운전조건을 다음과 같이 제시할 수 있었다. 삼각형 이론의 $m_2$와 $m_3$ 값은 각각 0.18, 1.0이고 스위칭 타임은 419 초이었다. 전산모사 결과 raffinate의 IgY 순도가 98.39%이고 두번째 싸이클에서 순도가 정상상태에 도달하였다.

• Journal of Animal Science and Technology
• /
• 제44권5호
• /
• pp.633-642
• /
• 2002

1. ETEC F41 균주로부터 분리한 섬모항원의 분자량은 29.5 kDa으로 나타났으며, western blot을 통하여 섬모항원임을 확인하였다. 2. 분리한 섬모항원의 농도를 50 ${\mu}g$/$m\ell$, 200 ${\mu}g$/$m\ell$, 600 ${\mu}g$/$m\ell$로 조정 후 산란계에 접종하였다. 이 후 ELISA법을 이용하여 난황의 항체역가를 측정한 결과 최고치가 320,000(antigen 50${\mu}g$/$m\ell$), 450,000(antigen 200${\mu}g$/$m\ell$), 320,000(anti- gen 600${\mu}g$/$m\ell$)으로 나타났다. 3. F41난황항체와 K88, K99, 987P 섬모항원과의 교차반응을 ELISA법을 이용하여 조사해본 결과 난황항체를 30,000배 희석 시 교차반응이 없었다. 4. 실험실조건하에서 난황항체의 항원결합능력을 조사한 결과, 동결건조한 WSF을 2${\sim}$4 mg/$m\ell$ 첨가 시 균체의 농도가 $10^9$ CFU/$m\ell$에서 $10^5$ CFU/$m\ell$로 급격하게 감소하였다.

• 원예과학기술지
• /
• 제32권4호
• /
• pp.544-549
• /
• 2014

Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.

• 대한수의학회지
• /
• 제44권3호
• /
• pp.415-420
• /
• 2004

Immunoprophylatic effect of IgY against B. bronchispetica was proven with 100% preventive rate in mice administrated with IgY with antibody titer 1:640~1:2,560. Intramuscular administration was more efficient than oral administration. This phenomenon was also observed in the therapeutic effects of IgY after challenge with B. bronchseptica in mice. In the field trials with the egg yolk antibodies from hens immunized with combined antigens with B. bronchiseptica and parvovirus, curing rates in dogs with severe clnical signs such as bloody diarrhea were 81.6% and 86.7% by intramuscular or subcutaneous administration of IgY, respectively. Safety of the antibodies in dogs was proven without any side effects such as vomiting, edema, fever, etc. by adminstration of double doses for 7 days. These results indicated that the egg yolk antibodies could be used as effective prevention and treatment of alimentary and respiratory diseases in dogs.

• 대한수의학회지
• /
• 제38권4호
• /
• pp.837-842
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제36권4호
• /
• pp.299-306
• /
• 2008

사람 췌장 리파아제는 췌장에서 합성되어 소장으로 분비된 후, 음식물속의 지방을 가수분해하는 소화효소이다. 췌장 리파아제 단백질은 촉매활성도메인과 코리파아제 결합도메인의 2개의 도메인으로 구성되어 있다 본 연구에서는 리파아제 전체단백질과 촉매활성도메인 및 코리파아제 결함도메인 유전자를 PCR 방법으로 증폭하여 발현벡터에 넣은 후, 대장균에서 발현시켜 얻은 3가지 재조합 단백질을 산란계에 3차례 주사하여 면역반응을 유도하였다. 난황단백질을 분리하여 항체가를 측정한 결과, 코리파아제 결합도메인의 항원성이 가장 높은 것으로 밝혀졌다. 또한, 돼지 췌장 리파아제의 활성저해 실험에서도 코리파아제 결합도메인에 의해 만들어진 난황항체가 지방분해활성 저해작용이 가장 큰 것으로 밝혀졌다. 코리파아제 결합도메인 난황항체가 첨가된 지방을 실험동물에게 급여하고 혈중 주요 성분을 분석한 결과, 코리파아제 결합도메인에 의한 난황항체를 섭취한 경우에 혈중 중성지방의 수치가 대조군에 비해서 유의적으로 감소했음을 화인하였다. 결론적으로 코리파아제 결합도메인을 항원단백질로 사용하여 얻은 난황항체가 생체 내에서 췌장 리파아제의 활성을 억제할 수 있으며 비만억제제로의 개발 가능성이 있음을 보여주었다.

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