Agrobacterium tumefaciens strain CP4 유래 5-enolpyruvylshikimate- 3-phosphate synthase (EPSPS) 유전자를 포함하는 유전자변형생물체 (Living modified organism, LMO)가 개발되었다. 이 같은 LMO는 국내 승인되어 사료용, 식품용, 가공용으로 이용 중이다. 간이면역 검사키트 개발을 위해서는 고효율의 단클론 항체 개발이 필수적이다. 본 연구에서는 대장균 BL21 (DE3)에서 재조합 CP4 EPSPS 단백질을 정제하였으며 SDS-PAGE와 MALDI-TOF MS 분석으로 단백질 특성을 분석하였다. 단클론 항체 제작은 ㈜앱 클론의 SOP 매뉴얼에 따라 진행하였다. 본 연구 결과 5개의 단클론 항체 클론 (2F2, 4B9, 6C11, 10A9, 10G9)를 확보하였다. 5종의 단클론 항체의 효율과 특이도 검정을 위해서 LM 면화 추출액을 이용한 western blotting 분석을 실시하였다. 모든 단클론 항체는 CP4 EPSPS를 함유하는 MON1445와 MON88913을 특이적으로 검출하였으며 비변형 면화 및 타종의 LM 면화에서는 검출되지 않았다. 이러한 결과들을 바탕으로 CP4 EPSPS 단클론 항체는 LMO 에 함유된 CP4 EPSPS 단백질을 타겟으로 항체 기반 검출법 개발에 활용될 것으로 사료된다.

Since the 2010 foot and mouth (FMD) epidemic, the Korean government has applied a FMD vaccination to pigs. A FMD vaccine is injected to pigs by intramuscular (IM) route. One of the drawbacks in FMD vaccine IM injection is that it would result in an abnormal meat on the injection spot. An abnormal meat due to FMD vaccine IM injection would cause economic loss to pig farmers. An intradermal (ID) injection would be an alternative method for FMD vaccination. The goal of current study was to compare the antibody formation rate between FMD vaccine IM injection and ID injection. The antibody formation rate was measured by the FMD serotype O vaccination percent inhibition (PI) value. In total 350 pigs (175 for FMD vaccine IM injection and 175 for FMD vaccine ID injection) were included in the study. In results, the PI values of FMD vaccine IM injection group were significantly higher than it of FMD vaccine ID injection group. However, the proportions of pigs with PI value was higher than 50, which is a legislative requirement for marketing pigs, for both FMD vaccine IM and ID injection groups at week 20 or 23 were not significantly different. The current results indicated that a FMD vaccine ID injection could be an alternative method of IM injection.

Rabies is a zoonotic disease that is caused by rabies virus and transmitted only in mammals. Domestic dogs are the most common reservoir of the virus, which is associated with more than 99% human deaths caused by dog-mediated rabies in the world. Rabies is one of the most fatal diseases, but it is fully preventable in animals by vaccination. Serological test of rabies virus antibody for samples collected from dogs and cats in Seoul during 2017–2019 was carried out in this study. To investigate antibody seroprevalence of rabies virus for dogs and cats, 2,769 serum samples were taken from 2,408 dogs and 361 cats in various regions of Seoul during 2017– 2019. Antibodies to rabies virus were detected by an indirect ELISA. Of 2,769 tested animal sera, 934 (33.7%) were positive; 886 (36.8%) of 2408 dogs and 48 (13.3%) of 361 cats. Of 990 companion animals, 547 (55.3%) was positive and 387 (21.8%) of 1779 stray animals was positive. These results indicate that antibody seroprevalence to rabies virus is still not enough to prevent rabies and rabies vaccination is required to enhance the antibody seroprevalence for rabies. To improve the situation, much public awareness and policy is needed to prevent the rabies. In addition, reducing stray animals and keeping companion animals from contact with wild animals are indispensable for the prevention of rabies.

ultrafiltration, diafiltr바이오의약품 중에서도 항체의약품의 수요가 증가함에 따라 항체를 상업용으로 개발하기 위한 연구가 활발히 진 행되고 있다. 항체의약품의 제조공정 중 downstream 공정은 의약품 성능에 직접적으로 영향을 미치기 때문에 중요하게 다뤄 지며, 본 총설은 그 중에서도 농축 및 제제화를 목적으로 행해지는 한외여과 및 정용여과 공정에서 사용되는 한외여과막의 주요 제품 및 공정 특성을 살펴보고, 최근 연구 동향에 대해 소개하고자 한다.ation, concentration, formulation, antibody-based therapeutics

In this study, antibody responses after vaccination against equine influenza were investigated among 1,591 horses in the Republic of Korea using the hemagglutination inhibition (HI) test. Equine influenza has not occurred since 2011 and a commercial vaccine against H3N8 has been used. The equine influenza virus, A/equine/South Africa/4/03 (H3N8), was used as the antigen in the HI assay. The mean seropositive rate was 90.5% in 2019. Except for stallion whose seropositive rate was 78.5%, all seropositive rates of other horse types were over 90%. Regionally, except for Gangwon-do and Jeju-do whose seropositive rates were 89.0% and 87.1%, all seropositive rates in other provinces were over 90%. In the future, more through vaccination against equine influenza needs to be done based on this investigation result.

Porphyromonas gingivalis is among the major etiological pathogens of chronic periodontitis. The virulence mechanisms of P. gingivalis is yet to be identified as its activity is largely unknown in actual disease process. The purpose of this study is to identify antigens of P. gingivalis expressed only in patients with chronic periodontitis using a unique immunoscreening technique. Change Mediated Antigen Technology (CMAT), an antibody-based screening technique, was used to identify virulence-associated proteins of P. gingivalis that are expressed only during infection stage in patients having chronic periodontitis. Out of 13,000 recombinant clones screened, 22 tested positive for reproducible reactivity with rabbit hyperimmune anti-sera prepared against dental plaque samples acquired from periodontitis patients. The DNA sequences of these 18 genes were determined. CMAT-identified protein antigens of P. gingivalis included proteins involved in energy metabolism and biosynthesis, heme and iron binding, drug resistance, specific enzyme activities, and unknown functions. Further analysis of these genes could result in a novel insight into the virulence mechanisms of P. gingivalis.

The purpose of this study was to investigate the understanding of immunization and antibody test by region and age. The questionnaire was conducted for those who raised their dogs for more than two years. The survey consisted of 500 people. SNS. The results showed that the perception of the reinforcement vaccination was more prevalent than Seoul / Gyeonggi province. However, in the Seoul / Gyeonggi area, the recognition and comprehension of the antibody test was higher and the participation rate by age was the highest in the 20s. In addition, the percentage of people who knew about vaccination was much higher, and the understanding of reinforcement vaccination showed a similar rate. The results of the questionnaire showed that the importance of antibody test is more important than antibody vaccination. In the case of a large number of antibodies, the reinforcement vaccination should be performed at the appropriate time after the antibody test.

The purpose of this study was to investigate the lesions of a mouse collagen antibody-induced arthritis (CAIA) model using fluorescence bioimaging and micro-computed tomography (micro-CT) and to compare it with histopathological examination. Twelve mice were randomly divided into three groups: group 1 (G1) as control, group 2 (G2) as fluorescence probe control and group 3 (G3) as collagen antibodyinduced arthritis. The mice of G3 intravenously received anti-type II collagen 5-clone antibody cocktail (2 mg/mouse) on day 0 and intraperitoneally received lipopolysaccharide (50 μg/mouse) on day 3. On the while, the mice of G1 and G2 received 0.9% saline in equal volumes at equivalent times. Fluorescence bioimaging and micro-CT analysis were carried out to assess arthritis. Treatment with the collagen antibody cocktail increased the paw thickness of mice compared to those in both the control and probe-treated groups. Fluorescence bioimaging using a near infrared imaging agent showed high intensity in the joints of collagen antibody- treated mice, whereas those of control mice showed no signal. Micro-CT analysis of the knee joints of collagen antibody-treated mice showed rough and irregular articular appearance, whereas those of control mice showed normal appearance. Histopathological examination of the knee joints of collagen antibody-treated mice revealed destruction of cartilage and bony structure, synovial hyperplasia and infiltration of inflammatory cells. No cartilage destruction or inflammation was observed in control or probe control mice. Taken together, it is concluded that analyses of fluorescent bioimaging made it possible to evaluate CAIA lesions, comparable with those by micro-CT and histopathological examination in mice.

For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without crossreaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

The object of this study was to evaluate Japanese encephalitis virus (JEV) antibody titer changes in broodmares and foals. Antibodies of 112 sera were detected by applying hemagglutination inhibition test. To the best of our knowledge, this is the first report that compares antibody titers of foals to that of their dams in order for evaluate optimal time of JEV vaccination. Most mares` antibody titers were variable. However, the highest titers in foals presented in their first month, and antibodies titers in all foals decreased gradually over time. This study provides important benchmarks that can be used to select optimum time JEV of vaccination.

Antibodies are used for preventing several infectious diseases in food animals. Though, antibiotics can deal with infection however, their widespread usage may cause developments of resistance and can also get transferred to humans through animal products. Therefore, production of antibodies against infectious agents in the egg yolk could be an interesting alternative. The present study was conducted with a focus on producing the antibody against Escherichia coli E68 and H28 in egg yolk of laying hens, which are believed to be a cause for diarrhea and beriberi in piglets. The result received from the experiment were promising and have shown efficiency of antibody production in the range of 2-7.3% in the groups which are infected with E68 only and the one which was infected with both strains. The outcome of present research has revealed the potentiality of egg yolk in production of antibody for laying hens, could open a new approach for production of antibodies to manage diarrhea and beriberi

Black queen cell virus (BQCV), one of the most prevalent viruse, causes the death of queen larvae and pupae. The RNA-dependent RNA polymerases (RdRPs) are central components in the life cycle of RNA viruses that catalyzes the replication of RNA from an RNA template without DNA stage. Inhibition of RdRP gene is importantly significant for application of monoclonal antibody generation as a diagnosis tool for identifying BQCV infection in honey bee..In this study, the presence of BQCV in honey bee samples was confirmed by PCR using BQCV F/R primer set to multiply of 700 bp DNA fragment. For ampification of BQCV Rdrp gene, a primer set attached BamHI/SalI restriction site was designed based on the best homogenization between BQCV RdRP sequences in NCBI, a PCR product containing BQCV RdRP gene with 1576 bp in length was amplified. Furthermore, BQCV RdRP gene will be cloned into pBlueXcm vector for future researches.

본 연구는 Sprague-Dawley 흰쥐 16마리를 이용하여 흰쥐 복강 및 피하지방 감소 후보 항체가 체중, in vivo 타장기 안전성 및 혈액성상 등 영양생리대사에 미치는 영향을 조사하고자 실시하였다. 무처리구(control), 비면역항체 처리구(NAb), 면양을 이용하여 기 개발된 흰쥐 복강(AAb) 및 피하지방 감소 후보항체(SAb)를 복강 주사하였을 때 주사 후 1주차에 SAb 처리구에서 사료 및 물 섭취량의 일시적인 감소가 나타났으나 그 이후에는 정상적으로 회복되는 것으로 보아 항체의 효과보다는 개체 차이에 기인하는 것으로 판단된다. 흰쥐 체중의 경우 항체 주사 처리 전․후 및 실험구 간 통계적인 유의차는 나타나지 않았다(p>0.05). SAb 처리구에서 나타난 주사 처리 후 1-2주 동안 일시적인 체중 감소 현상이 발생했으나, 통계적 유의성은 없었다(p>0.05). 후보 항체 주사 시 흰쥐 AAb 처리구는 control과 비교해서 생체 신장의 무게가 유의적으로 낮은 것으로 나타났다(p<0.05). 하지만, NAb 처리구의 신장 무게 역시 유의적인 감소를 보여(p<0.05) 결국 AAb 처리구의 신장 무게 감소는 주사에 의한 스트레스 또는 개체 간의 변이 때문인 것으로 생각된다. 신장을 제외하고는 후보 항체에 의한 주요 장기들(심장, 간장, 폐 및 비장)의 무게에는 실험구 간 유의적인 차이가 나타나지 않았다(p>0.05). 후보 항체에 의해 혈중 glucose, triglyceride 및 total cholesterol 농도에는 실험구 간 유의적인 변화가 나타나지 않았으며(p>0.05), 각 혈액대사물질의 농도는 일반적인 수준으로 나타났다. 이상의 결과를 볼 때, 본 연구에서 이용된 흰쥐AAb 및 SAb는 in vivo 영양생리대사에 부정적 영향을 미치지 않는 안전한 항체로 판단된다.

Gingival fibroblasts (GF) are the most abundant cell type in periodontal connective tissues, andhave distinct functional activities in the repair of periodontal tissues and in inflammatory periodontal diseases. Human gingival fibroblasts (hGF) can be used for periodontal tissue engineering. This study examined whether the alkaline phosphatase of hGF is enhanced by recombinant human BMP-4 and/or Anti human BMP-4 antibody. hGF was obtained from the excised gingival tissue of an implant patient undergoing 2nd surgery. The tissue was incubated at 37℃ in 5% CO2 and 95% humidity, and the cultivating media was changed every 2 days. The 2nd passage hGF cells were cultured in a medium containing Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and 1 X antibiotic antimycotic solution. The control hGF was cultured for 7 days without rhBMP-4/Anti human BMP-4 antibody. The experimental groups were cultured for 7 with BMP-4 (10 ng/ml) and/or Anti human BMP-4 antibody. This study evaluated the differentiation of hGF to osteoblasts using alkaline phosphatase assay. In the experimental groups, the hGF showed abundant positive ALP staining. Among the experimental groups, the experimental group 3 (mixture of rhBMP-4 (20ng/㎖) and Anti human BMP-4 antibody (50000ng/㎖) showed most abundant positive ALP staining. In the control group, the hGF showed weak positive ALP staining. Overall, these results suggest that the ALP expression of hGF can enhanced by rhBMP-4 or mixture of rhBMP-4/ anti human BMP-4 antibody.

We prepared the polyclonal antibody anti-20α-hydroxysteroid dehydrogenase (anti-20α-HSD) against the recombinant full-length protein bovine 20α-HSD in Escherichia coli. The specificity of anti-20α-HSD was demonstrated using Chinese hamster ovary (CHO) cells transfected with recombinant bovine 20α-HSD and bovine placental tissues. According to western blot analysis, anti-20α-HSD specifically recognizes the 37-kDa protein bovine 20α-HSD. The protein is not present in untransfected CHO cells. Anti-20α-HSD also recognizes a specific protein in the ovaries and placenta of other animals. Immunostaining was used to detect expression of bovine 20α-HSD protein in the cultured luteal cells during the estrous cycle later.

The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

Equine coital exanthema (ECE) caused by equine herpes virus type 3 (EHV-3) is a sexually transmitted disease which is resulted in the failure of mating, declination of horse productivity and finally economic loss in horse industry. In this thesis, diagnosis studies on ECE have been performed in order to develop a serological diagnosis method of enzyme-linked immunosorbent assay (ELISA). ELISA has been developed to detect antibodies against EHV-3. Whole EHV-3 viruses were purified from cell culture and coated on ELISA plate, which successfully captured by anti-EHV-3 antibody in serum samples. The positive cut-off value of developed ELISA was 0.334 at OD405 using 15 negative control horse sera. The positive rate of 20 sera from ECE positive horses by PCR was 65% and the positive rate of 12 sera from ECE negative horses by PCR was 25%. The positive rate of 72 sera from horses showing clinical signs was 59.7% and the positive rate of 72 sera from horses showing no clinical signs was 13.9%. The correlation of serum positivity between broodmares and their sucklings was analysed using 12 pairs of cases (y=0.5418x—0.0158, R2= 0.4931). The data suggested that specific antibody against EHV-3 from broodmares might be transferred to their sucklings by nursing. When compared with the results of PCR, the sensitivity and specificity of ELISA were 65%, 81.8% respectively.