• Archives of Pharmacal Research
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• 제27권9호
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• pp.961-967
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• 2004

The development of an antibody labeling method with $^{99m}$Tc is important for cancer imaging. Most bifunctional chelate methods for $^{99m}$Tc labeling of antibody incorporate a $^{99m}$Tc chelator through a linkage to lysine residue. In the present study, a novel site-specific $^{99m}$Tc labeling method at carbohydrate side chain in the Fc region of 2 antibodies (T101 and rabbit anti-human serum albumin antibody (RPAb)) using dihydrazinophthalazine (DHZ) which has 2 hydrazino groups was developed. The antibodies were oxidized with sodium periodate to pro-duce aldehyde on the Fc region. Then, one hydrazine group of DHZ was conjugated with an aldehyde group of antibody through the formation of a hydrazone. The other hydrazine group was used for labeling with $^{99m}$Tc. The number of conjugated DHZ was 1.7 per antibody. $^{99m}$Tc labeling efficiency was 46-85％ for T101 and 67∼87％ for RPAb. Indirect labeling with DHZ conjugated antibodies showed higher stability than direct labeling with reduced antibodies. High immunoreactivities were conserved for both indirectly and directly labeled antibodies. A biodistribution study found high blood activity related to directly labeled T1 01 at early time point as well as low liver activity due to indirectly labeled T101 at later time point. However, these findings do not affect practical use. No significantly different biodistribution was observed in the other organs. The research concluded that DHZ can be used as a site-specific bifunctional chelating agent for labeling antibody with $^{99m}$Tc. Moreover, $^{99m}$Tc labeled antibody via DHZ was found to have excellent chemical and biological properties for nuclear medicine imaging.edicine imaging.

• 한국동물위생학회지
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• 제32권3호
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• pp.201-207
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• 2009

Porcine transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV) and rotaviruses are considered as the most important causative agents of diarrhea in piglets. The study established 3 method vaccination programs to prevent PEDV. A (LL)group inoculated twice vaccinations on 2-weeks-interval during the late term of pregnant sows with PEDV live vaccine. The B (LKK) group was applied that one time single PEDV live vaccine at the pre-mate followed by the TGEV PEDV combined inactivated vaccine (twice vaccination on 2-weeks interval at the third-trimester). C (KK) group was applied to sow which inoculated twice vaccination on 2-weeks-interval during the late term of pregnant sows with by the TGEV, PEDV combined inactivated vaccine. As the result of SN test on sows in the pig farm before vaccination, antibody titers was showed 9/45 (20.0%). By comparison with the serum neutralizing antibody titers against PEDV of the vaccination programs after PEDV of the vaccination, A group and B group vaccination method was higher than those of C group in sows. In the piglets up to 2 weeks of age, A group was showed antibody titers of 17/22 (81.8%) that showed 2-128, and B group was showed antibody titers of 30/37 (81.1%) that showed 2-512, and C group was showed antibody titers of 14/28 (50.0%) that showed 2-32. On the other hand, PEDV antibody titers were tested for the survey. As the results of SN test, Aujeszky's disease survey in 54 pig farms from november 2005 to august 2006, antibody titers of 47/286 (16.4%) showed above 2. Five breeding farms were antibody titers of 38/77 (49.4%), Wanggung zone farms antibody titers of 59/85 (69.4%). In pigs farms vaccinated the first of twice PEDV live vaccine, and after 6 month, the second of twice TGEV PEDV combined inactivated vaccine (LLKK, 256-1024 titer) method was higher than those of vaccinated twice the early term of pregnant, and twice the late term of pregnant sows of PEDV live vaccine (LLLL, 32 titer).

• Parasites, Hosts and Diseases
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• 제29권4호
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• pp.389-396
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• 1991

인체아니사키스증은 우리나라에서도 드물지 않게 발생하나 과거에는 수술에 의한 충체의 확인만이 진단이자 치료이어서 진단에 많은 어려움을 겪었다. 그러나 위내시경이 진단과 치료에 이용되면서 많은 중례의 발견을 가져오게 되었다. 그러나 증상이 심하지 않을 경우나 위내시경의 삽입이 어려운 장(장) 아니사키중 또는 만성 아니사키스중을 진단하기 위해서는 혈청학적 방법이 필요하다. 이 연구는 아니사키스중의 혈청학적 진단연구를 위해 붕장어에서 채집한 유충을 토끼에 10마리섹 감염시키고 IsM, IgG 항체의 출현시기와 변화양상을 ELISA와 SDS-PAGE/immunoblot으로 관찰하였다. 그 결과는 다음과 같다. 1. ELISA에 의하여 측정한 19M 항체는 감염 6일부터 증가하여 11일째 최고치에 도달한 후 35일 이후에는 감 염 전의 수준으로 저하하였고 IgG 항체가는 감염 6일부터 증가하여 26일째 최고치에 도달한 후 95일까지 감소하는 양상을 보였다. 2. 7.5∼15% SDS-PAGE에서 아니사키스추출액은 최소 41개 이상의 단백분회 을 나타냈으며 195, 145, 110, 104, 73, 69, 49, 42.5, 40, 34, 30, 24, 20, 16, 14 kDa가 주 분획이었다. 3. Immunoblot에 의해 IgM 항체에 반응하는 주 band는 168, 95, 74, 64, 51, 47, 34 kDa의 5개 band이었다. 4. IgG 항체와 반응하는 항원 분획은 11개 이상 관찰되었고 그 중 168, 92, 85, 64, 58, 52, 42, 40 kDa가 강한 반응을 보였다. 5. 아니사키스추출액 항원과 다른 기생충성 질환자 혈청과의 교차반응은 관찰할 수 없었다.

• 한국동물위생학회지
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• 제14권1호
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• pp.1-5
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• 1991

A serological survey for antibody of Leptospira spp. in canine was carried out from March to September, 1989 in Seoul. 182 serums collected from animal hospitals and keeping were collected and these were performed by using 12 different living antigens. In the microscopic agglutination test(MAT), being partial agglutination reaction at a serum dilution of 1：200 or over, we recorded it as positive. These results were compared with the species, sex and general conditions of canines, the areas and types of animal keeping. The results were summarized as followed ; 1. We detected the antibodies L. grippotyphosa 1 and L. icteroheamorrhagiae 1 in A area(total 48 heads ), L. canicola 1 and L. icterohaemorrhagiae 4 in B area(total 52 heads), L. hardjo 1 and L. icterohaemorrhagiae 2 in C area (total 32 heads), L. icterohaemorrhagiae 1 in D area(total 23 heads) L. grippotyposa 1 and L. icterohaemorrhagiae 2 in E area(total 27 heads) by MAT. There were positives for L. canicola 1, L. grippotyposa 2, L. hardjo 1 and L. icterohaemorrhagiae 10 in 5 areas by MAT. 2. The deteclive rate of leptospiral antibody in Jindo canine was 17.6% (3) among 17, Mixed 4.4% (4) among 90 and Exotic 9.3% (7) among 75 heads. 3. The Male(91 heads) was positive for 8.7%(8) and the female(91 heads) was positive for 6.5%(6). 4. In the vaccination, positive rate was 10.3% (7) among 55 heads, and in the unvaccination, positive rate was 5.5%(7) among 127 heads.

• 대한수의학회지
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• 제32권3호
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• pp.377-380
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• 1992

보르나병 바이러스 혈청학적 진단법에 있어서 새로 개발된 세포효소면역법의 항체검출에 대한 평가를 위하여 지금까지 주로 사용되어진 간접형광항체법과 네 종류의 실험동물에서 서로 비교하였다. 모든 동물에서 간접형광항체법의 역가와 세포효소면역반응치 사이에 상관계수가 모두 0.8 이상으로 고도의 유의성이 인정되었고 두 진단법의 일치율은 한 희석단계 이하로서 재현성이 아주 좋았다. 세포효소면역반응법은 보르나병 바이러스의 혈청역학적 조사 및 병인기전 연구에 유용하게 이용될 수 있을 것으로 생각된다.

• Nuclear Engineering and Technology
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• 제12권2호
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• pp.121-126
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• 1980

효과적인 저농도 인슈린 방사면역 측정법 확립 및 표지인슈린의 면역학적 환성과 탄소분말에 대한 흡착력의 상관관계를 밝히기 위하여 이들 두 다른 기준에 기초를 둔 두가지의 최적표지 인슈린선정방법을 비교한결과 탄소분말흡착에 의한 방법은 부적당함을 알 수 있었다. 일반적으로 표지인슈린의 방사능, 항체결합능, 탄소분말흡착능등은 서로 일치하지 않았다. 따라서 측정신뢰도를 높이기 위해서는 매 표지때마다 항체결합능으로 최적표지 인슈린을 선정해야 하며 탄소분말 흡착력으로 선정하면 안된다고 본다. 항체결합능에 따라 선정된 표지 인슈린을 써서 대조혈청중의 인슈린량이 정화히 측정되었으며 정온유지시간을 96시간으로 연장함으로써 5 $\mu$U/ml 하의 표준 인슈린투여 량에 대해서도 매우 예리한 응답곡선을 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.95-98
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• 2000

An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.268-273
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• 2015

The fluorescent-antibody-to-membrane-antigen (FAMA) test is regarded as the "gold standard" to detect protective antibodies to varicella-zoster virus (VZV) because of its high sensitivity and specificity. Because the classic FAMA test uses an infectious virus for detection of antibodies to VZV, it is labor-intensive, and also requires special equipment for handling the virus. For this reason, we attempted to develop a simple and safe FAMA assay. Because VZV glycoprotein E (gE) is one of the major VZV glycoproteins, we used the gE protein for the FAMA test (gE FAMA). Here, we demonstrate that overexpression of gE in HEK293T cells can be used to measure antibodies in human serum, and that gE FAMA titers are closely correlated with gpEIA ELISA data. These results indicate that our gE FAMA test has the potential to measure antibodies to VZV.

• 한국동물위생학회지
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• 제29권3호
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• pp.365-376
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• 2006

To identify immune response of leukocytes in peripheral blood of cattle vaccinated with an attenuated live Akabane virus vaccine, leukocytes were reacted with monoclonal antibodies which are specific to bovine lymphocyte surface antigens and assayed by the flow cytometry. Serum neutralizing (SN) test was used to measure antibody titers after vaccination, SN antibody was appeared to 7 days post-vaccination (PV) and 2-8 antibody titers were observed in 14 days PV. Proportion of $CD8^-$ MHC $class II^+$ expressing cells were rapidly increased at 3 days PV. $CD8^+$ MHC $class II^-$ cells were increased at 7 days PV. $CD4^+CD8^-,\;WC^+CD4^-,\;CD4^+CD8^+,\;WC1^-CD4^+, \;WC1^-CD8^+$, and $CD4^-CD8^+$ cells were highly increased at 3, 3, 7, 7, 14, 14 days PV, respectively.

• 한국동물위생학회지
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• 제42권4호
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• pp.237-243
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• 2019

Avian reovirus (ARV) is the pathogenic agent of tenosynovitis and malabsorption syndrome in broiler, which has caused significant economical losses due to poor feeder efficiency and stunting. In order to determine the prevalence of ARV infection in poultry farms in Jeonbuk province, Korea, we performed a surveillance study by testing 179 cecal samples from 131 broiler farms for virus detection, and 1,181 serum samples from 33 broiler farms (n=292) and 22 broiler breeder farms (n=1,525) for antibody detection in the province. Virological examination using RT-PCR showed that ARV were detected in 26.0% of the tested farms (34/131),with the highest positive rates in broilers of 6 days old or more in summer season. In serological test using ELISA, broiler and broiler breeder farms examined were all ARV antibody positive. In broiler, the positive rate and antibody titers showed a tendency to decrease with age in contrast to those of broiler breeders. Our results indicate that ARV is ubiquitous in broilers and broiler breeders in the province.