• Parasites, Hosts and Diseases
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• 제60권2호
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• pp.143-147
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• 2022

Acanthamoeba keratitis (AK) is a rare ocular disease, but it is a painful and sight-threatening infectious disease. Early diagnosis and adequate treatment are necessary to prevent serious complications. While AK is frequently diagnosis via several PCR assays or Acanthamoeba-specific antibodies, a more specific and effective diagnostic method is required. This study described the production of a polyclonal peptide antibody against the periplasmic binding protein (PBP) of A. castellanii and investigated its diagnostic potential. Western blot analysis showed that the PBP antibody specifically reacted with the cell lysates of A. castellanii. However, the PBP antibody did not interact with human corneal epithelial (HCE) cells and the other 3 major causative agents of keratitis. Immunocytochemistry (ICC) results revealed the specific detection of A. castellanii trophozoites and cysts by PBP antibodies when A. castellanii were co-cultured with HCE cells. PBP antibody specificity was further confirmed by co-culture of A. castellanii trophozoites with F. solani, S. aureus, and P. aeruginosa via ICC. The PBP antibody specifically reacted with the trophozoites and cysts of A. polyphaga, A. hatchetti, A. culbertsoni, A. royreba, and A. healyi, thus demonstrated its genus-specific nature. These results showed that the PBP polyclonal peptide antibody of A. castellanii could specifically detect several species of Acanthamoeba, contributing to the development of an effective antibody-based AK diagnostics.

• 한국동물위생학회지
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• 제26권1호
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• pp.51-56
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• 2003

Avian pneumoviros(APV), also known as avian rhinotracheitis virus(ARTV), affects both turkeys and chickens and is known to be the primary causative agent of turkey rhinotracheitis (TRT). The aim of this study was to establish the presence or absence of antibodies to avian pneumovirus in the commercial poultry population of Korea. For this purpose, chicken serum samples were obtained and tested by an enzyme-linked immunosorbent assay (ELISA). The tested serum was collected in laying hens with reduction of egg production or normal in Gyeongbuk province. A total of 184 sera representing 42 different poultry farms of the Gyeongbuk region of Korea were included in this study. Laying hens of 16 different farms with reduction of egg production and laying hens of 26 different farms with clinically healthy at the time of serum sampling were considered positive to antibody against APV. In the farms with reduction of egg production, positive farm to antibody against avian pneumovirus were 11 of 16 different farms(68.8%) and positive sera were 47(58.8%) of 80 different serum. In the farms with clinically healthy flock, positive farm to antibody against avian pneumovirus were 12(46.2%) of 26 different farms and positive serum sample were 39(37.5%) of 104 different sera. According to the results tested to 42 different farms in 14 city, 8 of 14 city have flocks with antibody positive laying hens against APV, 1 of 14 city have antibody suspicious and 5 of 14 city shown antibody negative, respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제28권3호
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• pp.323-327
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• 2015

Wattle length, width, and area were measured to classify bilateral asymmetries in four lines of chickens. The lines were the S26 generation of White Leghorns selected for high (HAS) or low (LAS) response to sheep red blood cells and sublines in which selection had been relaxed for three generations (high antibody relaxed [HAR] and low antibody relaxed [LAR]). Antibody titers (AB) were greater for HAS than for HAR with both greater than for LAS and LAR which while different for males did not differ for females. The low antibody lines were heavier and reached sexual maturity at younger age than the high antibody lines. In general, wattle length, width, and area were greater in the low than high antibody lines. In 24 comparisons for bilaterality 18 exhibited fluctuating asymmetry and 6 exhibited directional asymmetry with 5 of the 6 being for wattle length. There was not a clear pattern for changes in degree of asymmetry when selection was relaxed for 3 generations. For females, the relative asymmetry (RA) of wattle area was larger ($p{\leq}0.05$) for HAR than for LAR and not different from the selected lines and relaxed lines. There were no differences among lines for RA of wattle length and width of females and wattle length, width, and area of males.

• BMB Reports
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• 제32권5호
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• pp.474-479
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• 1999

Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.93-93
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• 1997

Analysis of protein is often frustrated by the inability to isolate large amounts of purified protein from a native source. To overcome this problem, fusion protein expression systems such as pGEX system have been widely used. Using pGEX system, the desired protein could be easily obtained in a large amount in E. coli, and then the fusion protein could be used for the study of the function of the given protein. To analyze and purify the GST fusion protein, anti-GST antibody could be used as one of the system of choice. However, the production and characterization of monoclonal anti-GST antibody has not been studied extensively yet. To produce monoclonal anti-GST antibody, GST was purified from E. coli transformed with pGEX-cs, one of the pGEX system and was used as an antigen. The monoclonal antibody was produced by fusion of the immunized spleen cells with SP2-0 myeloma cells. The antibody was characterized by ELISA, western blotting, etc. The monoclonal antibody produced in this study (mAb-GSTA) showed strong and specific immunoreactivity against not only GST but also GST-fusion proteins. Also, mAb-GSTA was successfully used for the immunoaffinity purification of the GST ${\beta}$-Rc.-third intracellular-loop fusion protein. The results of the present study suggest that mAb-GSTA may be used for the identification and purification of GST fusion proteins.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.141-144
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• 2006

An antibody layer was fabricated to detect Escherichia coli O157:H7. The micropattern of 16-mercaptohexadecanoic acid (16-MHDA) as alkylthiolate was formed on the gold surface by using the PDMS stamp with microcontact printing $({\mu}CP)$ techniques. In order to form antibody patterns on the template, protein G was chemically bound to the 16-MHDA patterns, and antibody was adsorbed on a self-assembled protein G layer. The formation of the 16-MHDA micropattern, self-assembled protein G layer and antibody pattern on Au substrate was confirmed by surface plasmon resonance (SPR) spectroscopy. Finally, the micropatterning method was applied to fabricate the antibody probe for detection of E. coli O157:H7, and monitoring of antigen by using this probe was successfully achieved.

• Journal of Biosystems Engineering
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• 제28권2호
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• pp.167-172
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• 2003

This study was performed to fabricate a batch-type SPR biosensing system using a thiolated E. coli antibody coupling, and to explore the feasibility of real-time detection of E. coii in a stagnant sample solution. In advance. “O” and “K” antigenic serotype E. coli antibodies were thiolated with sulfo-LC-SPDP and dithiothreitol, and immobilized by chemisorption in the gold surface of compact SPR sensors. When the SPR biosensor immobilized with E. coli antibody monitored a E. coli solution, it took 3 to 5 min to stabilize. The SPR biosensing system developed in this study was able to detect E. coli in the range above 10$^4$ CFU/mL at the 0.05 significant level. Also, the SPR biosensor had possibility to significantly detect E. coli in the range of 10$^2$ to 10$^4$ CFU/mL in E. coli solutions. Meanwhile, when the SPR biosensor immobilized with 5. coli antibody was cleaned with NaOH solutions, its ability to detect E. coli largely decreased due to wash-out of the immobilized antibody. In order to reuse the SPR sensor, it should be antibody-immobilized newly.

• 약학회지
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• 제59권4호
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• pp.170-176
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• 2015

In vaccine development, the major points may be induction of effective and increased levels of antibody production. This is especially the case when the antigenic sources are carbohydrates. For many years, thus, we have researched various types of formulations such as liposomal and conjugate vaccines. However, the fastidious formulation process and high costs are a problem. For this reason, there is currently a focus on utilizing immunoadjuvants. In this present study, we tested if platycodin D (PLD) from Platycodon Radix have immunoadjuvant activity against the cell wall of Candida albicans (CACW). The resulting data showed that in the murine model of antibody production, CACW combined with PLD [CACW/PLD/IFA] increased the production of antibodies specific to C. albicans when compared to the antibody production by [CACW/IFA]-induction, which was used as a negative control (P<0.05). In the case of [CACW/PLD/IFA], the antibody production was 1.4 times as that of the CFA. In addition, formulations containing either had a prolonged antibody inducing activity maintaining the initial titers of antibody as compared to the CFA formula. Cytokine profiling with the antisera displayed that the PLD produced both Th1 and Th2 immunoresponses, but Th1 dominant was much greater (P<0.05). Furthermore, [CACW/PLD/IFA] formula enhanced resistance of mice against disseminated candidiasis, whereas the CFA had no such effect. In conclusion, PLD has an immunologic activity, which is protective against the disease. Thus, PLD can be a goof candidate for a new immunoadjuvant in development of the fungal vaccine.

• 대한수의학회지
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• 제19권1호
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• pp.45-51
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• 1979

The present study was undertaken to observe the immune status of breeding hens and laying hens against Newcastle disease (ND). The methods of extraction of hemagglutination inhibition (HI) antibody from egg yolk, the detection of HI antibody in egg albumen and the correlation between HI antibody titers in maternal sera and egg yolks were discussed. For the purposes of these experiments, 9 flocks of breeding hens and 16 flocks of laying hens immunized against Newcastle disease virus were investigated. The vaccination program of tested flocks was 3-3-3 or 4-4-4 in general. The results obtained are summerized as follows: Freezing-thawing was the best method far antibody extraction from egg yolk for HI test. The HI antibody against NDV was found in egg albumen (geometric mean, 4.5), but lower than that found in egg yolk (32.1). The geometric mean of HI antibody titers of egg yolks (84.1) was higher than that of maternal sera (68.4) and day-old chicken sera (25.3). There was correlation between HI antibody titers of maternal sera(Y) and those of egg yolks(X). The coefficient correlation was r=0.63, and the line of regression of Y on X was $\hat{Y}$=35.91+0.35X.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.465-470
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• 2009

The chicken Mx gene has been regarded as a candidate gene for resistance to avian influenza virus (AIV). In this study, three groups of chickens with homozygotes (AA, GG) and heterozygotes (AG) of the resistant (A) and susceptible alleles (G) to AIV of the Mx gene were constructed from a line of dwarf egg-type chickens. These chickens were not examined for their resistant activities to AIV because the differential resistance had only been detected in vitro. The birds of the three groups were vaccinated with inactivated H5N2 AIV vaccine and the level of hemagglutination inhibition (HI) antibody to AIV was detected. The association between disease resistant activity to AIV and antibody response to AIV vaccination in the three groups was analyzed. The chickens with homozygous resistant allele A showed the lowest antibody levels, whereas the heterozygous chickens (AG) presented the highest antibody level after the boosting vaccination, which indicates that the efficiency of artificial selection on the resistant allele of Mx gene will be compromised since the homozygotes of the allele presented the weakest antibody response to the corresponding vaccine.