• Korean Journal of Veterinary Service
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• v.14 no.1
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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.

• Korean Journal of Veterinary Service
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• v.16 no.1
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• pp.34-40
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• 1993

The present study was conducted to investigate the isolation frequency, biochemical prop erties and antimicrobial susceptibility of B. bronchiseptica isolated from slaughtered pigs during the period from March to December, 1992. In Kyeonggi province. A serological survey for antibody of B. bronchiseptica in 200 slaughtered pigs was carried out by agglutination and tetrazolium reduction methods. The results were summarized as follows ; 1. From 80 slaughtered pigs, 27(33.8%) case were isolated and all isolate strains were resistant to Penicillin, Streptomycin, Chloramphenicol, Tetracycline and Ampicllin, while the majority of them were susceptible to Gentamicin, Cloxacin, Colistin, Neomycin, and Kanamycin. 2. Incidence of B. bronchiseptica antibody in 200 slaughtered pigs were measured by agglutination and tetrazolium reduction methods. Agglutination method was shown 38 (19%) of 200 with a titer of below 1：20 and 20(10%) of 200 with a titer of above 1：640. Tetrazolium reduction method was observed 33(16.5%) of 200 with a titer of below 1 : 20 and 32(15%) of 200 with a titer of above 1：640. 3. LSD analysis indicated that the difference of the responses between agglutination test and tetrazolium reduction test was not significant.

• BMB Reports
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• v.29 no.5
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• pp.462-467
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• 1996

Acetolactate synthase (ALS, EC 4.1.3.18) is the first common enzyme in the biosynthesis of leucine, isoleucine, and valine. It is the target enzyme for several classes of herbicides, including the sulfonylureas, the imidazolinones, the mazolopyrimidines, the pyrimidyl-oxy-benzoates, the pyrimidyl-thio-benzens, and the 4,6-dimethoxypyrimidines. An amino-terminal fragment of the sulfonylurea-resistant ALS gene (SurB) from Nicotiana tabaccum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS1 was used to transform Escherichia coli strain BL21, and the tobacco ALS was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). Polyclonal antibodies against the fusion product (GST-ALS) were produced, and the sensitivity of GST-ALS with the rabbit anti-GST-ALS IgG was up to 50 ng. This antibody was used for Western blot analysis of the partially purified ALS from barley shoots. The results suggest that the polyclonal antibody produced in this study can be used to detect plant ALS.

• Biomedical Science Letters
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• v.9 no.3
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• pp.111-121
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• 2003

Recombinant single chain Fv (scFv) antibodies offer many advantages over mouse monoclonal antibodies such as faster clearance from blood, improved tumor localization, reduced human anti-mouse antibody (HAMA) response, and the availability to manipulate the scFv through genetic approaches. The recombinant phage display was constructed using lym-l hybridoma cells as a source of genetic starting material. mRNA was isolated from the corresponding antibodies hybridoma cells. VH and VL cDNA were amplified with RT-PCR and linked with ScFv by linker DNA to form ScFv DNA, which then were inserted into phagemid pCANTAB5E. The phage of positive clones selected with tube containing raji lymphoma cell and infected by competent E. coli HB2151 to express soluble scFv. The scFv lym-l was secreted into the cytosol and culture supernatant and shown to be of expected size (approximately 32 kDa) by western blot. An active scFv lym-l could be produced in E. coli with soluble form and high yield from hybridoma cell line, using phage display system. Immunoreactivity indicated that scFv lym1 showed a potential biding affinity against the raji lymphoma cell as its parental antibody (intact lym-l Ab).

• Animal cells and systems
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• v.8 no.2
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• pp.117-123
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• 2004

The purpose of this study was to analyze inhibitory effects of anti-4-1BB monoclonal antibody on melanoma metastasis The 4-1BB (CD137) T cell molecule is a member of the TNF receptor family and its activation by either 4-1BB ligand or antibody induces T cell activation and growth. In the present study, administration of anti-4-1BB mAb induced inhibition of melanoma metastasis. Agonistic anti-4-1BB mAb induced not only CD$8^+$4-1BBT cells but also CD$8^+$IFN-${\gamma}$$^{+}$ T cell population. In the presence of anti-CD3 antibody, lymphocytes produced high levels of IFN-${\gamma}$ and low levels of IL-4 in anti-4-1BB mAb treated group. Exposure of melanoma cells to IFN-${\gamma}$ induced expression of MHC-I molecules. Thus, the increase in number of CD$8^+$T cells and enhanced MHC-I expression on B16F10 cells by augmented IFN-${\gamma}$ production in response to anti-4-1BB mAb may result in suppression of tumor growth and metastasis.s.

• Journal of Microbiology and Biotechnology
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• v.32 no.12
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• pp.1615-1621
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• 2022

Tissue regeneration is the ultimate treatment for many degenerative diseases, however, repair and regeneration of damaged organs or tissues remains a challenge. Previously, we showed that B1 Ab and H3 Ab induce stem cells to differentiate into microglia and brown adipocyte-like cells, while trafficking to the brain and heart, respectively. Here, we present data showing that another selected agonist antibody, P1 antibody, induces the migration of cells to the pancreatic islets and differentiates human stem cells into beta-like cells. Interestingly, our results suggest the purified P1 Ab induces beta-like cells from fresh, human CD34⁺ hematopoietic stem cells and mouse bone marrow. In addition, stem cells with P1 Ab bound to expressed periostin (POSTN), an extracellular matrix protein that regulates tissue remodeling, selectively migrate to mouse pancreatic islets. Thus, these results confirm that our in vivo selection system can be used to identify antibodies from our library which are capable of inducing stem cell differentiation and cell migration to select tissues for the purpose of regenerating and remodeling damaged organ systems.

• Korean Journal of Veterinary Service
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• v.43 no.4
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• pp.217-225
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• 2020

This study was conducted to survey pathogens of canine coronavirus (CCV), canine distemper virus (CDV), canine influenza virus (CIV), canine parvovirus (CPV), severe fever with thrombocytopenia syndrome virus (SFTSV), Dirofilaria (D.) immitis, Giardia and antibodies against Anaplasma (A.) phagocytophilum, Borrelia (B.) burgdorferi, Brucella (B.) canis and Ehrlichia (E.) canis among stray dogs in Gyeonggi province. We collected 271 feces, 291 bloods, 311 nasal and ocular swab samples from 311 of dogs in the Gyeonggi province assistance dogs sharing center from January to December, 2019. Among canine infectious disease pathogens, Giardia was highly detected in 46/271 (17.0%) samples. Subsequently, CCV 10.3% (28/271), D. immitis 8.2% (24/291), CPV 4.1% (11/271), CDV 1.0% (3/311), A. phagocytophilum (antibody) 0.3% (1/291), E. canis (antibody) 0.3% (1/291) were detected. Based on the results, this study is expected to provide a useful reference for disease control and management of stray dogs.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.1
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• pp.57-71
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• 2009

Objectives : The aim of this study was to know the immunity reponse of Esubwhaltong-tang(hereafter referred to ESWTT) to rheumatoid arthrits in CIA(collagen induced arthritis) mice. Methods : For this purpose, ESWTT was orally administerd to mice with arthritis induced by collagen II and then value of immunocyte in paw joint, cytokine(IL-6, $TNF-{\alpha}$), rheumatoid factor(IgG and IgM) and collagen II specific antibody in the serum were measured. Results : 1. The cytotoxicity was not shown on hFLSs and liver. 2. Marginal erosion, necrotic chodrocytes, cartilage and bone degradation were improved in histological section of paw joints from CIA mice(ESWTT extract administration group). 3. Total cell number of paw joint in CIA mice(ESWTT extract administration group) was decreased significantly. 4. The absolute number of CD3+, CD3+/CD69+, CD4+, CD4+/CD25+, CD49b+, CD3+/CD49b+ cells in CIA mice(ESWTT extract administration group) were decreased significantly. 5. The levels of IL-6 and $TNF-{\alpha}$ in the serum of CIA mice(ESWTT extract administration group) were decreased significantly. 6. The levels of total IgG and IgM in the serum of CIA mice(ESWTT extract administration group) were decreased significantly. 7. The level of collagen II specific antibody in the serum of CIA mice(ESWTT extract administration group) was decreased significantly. Conclusions : Comparison of the results for this study showed that ESWTT had immunomodulatory effects of suppressing. So we expect that ESWTT could be used as an effective drugs for not only rheumatoid arthritis but also auto-immune disease.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.148-156
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• 2021

Single-chain antibodies against epidermal growth factor receptor variant III (EGFRvIII) are potentially promising agents for developing antibody-based cancer treatment strategies. We described in our previous study the successful expression of an anti-EGFRvIII scFv antibody in Escherichia coli. However, we could also observe the formation of insoluble aggregates in the periplasmic space, limiting the production yield of the active product. In the present study, we investigated the mechanisms by which growth conditions could affect the expression of the soluble anti-EGFRvIII scFv antibody in small-scale E. coli NiCo21(DE3) cultures, attempting to maximize production. The secreted scFv molecules were purified using Ni-NTA magnetic beads and protein characterization was performed using SDS-PAGE and western blot analyses. We used the ImageJ software for protein quantification and determined the antigen-binding activity of the scFv antibody against the EGFRvIII protein. Our results showed that the highest percentage of soluble scFv expression could be achieved under culture conditions that combined low IPTG concentration (0.1 mM), low growth temperature (18℃), and large culture dish surface area. We found moderate-yield soluble scFv production in the culture medium after lactose-mediated induction, which was also beneficial for downstream protein processing. These findings were confirmed by conducting western blot analysis, indicating that the soluble, approximately 30-kDa scFv molecule was localized in the periplasm and the extracellular space. Moreover, the antigen-binding assay confirmed the scFv affinity against the EGFRvIII antigen. In conclusion, our study reveals that low-speed protein expression is preferable to obtain more soluble anti-EGFRvIII scFv protein in an E. coli expression system.

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.445-451
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• 2021

In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.