• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.147-151
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• 2004

Antigenicities of ovomucoid (OM) and ovalbumin (OA) in chicken egg white (EW) before and after NaOH, heat, and pretense treatments were examined by competitive indirect enzyme-linked immunosorbent assay (ciELISA), using rabbit anti-OM and-OA antibodies, Enzymatic hydrolysis of EW did not effectively reduce antigenicity of OM, whereas that of OA was decreased to 1/5,000-1/100,000 by treatment of plant-derived or microbial pretenses. Heat treatment below $100^{\circ}C$ for 30min did not decrease antigenicity of OM, whereas that of OA in heated EW increased maximally to 100 times, Antigenicity of OM in EW effectively decreased by NaOH treatment, disappearing at over 1% NaOH, whereas that of OA increased. Additional heat treatment of NaOH-treated EW at $70^{\circ}C$ for 15min slightly reduced antigenicities of OM and OA.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.677-681
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• 2003

Currently, both the indirect fluorescent antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA) have been used to detect Neospora caninum antibodies. Several factors such as the buffers, the conjugate, the pattern of fluorescence, and the cross reactivity with other apicornplexan protozoan, may result in poorly correlated data. The present study was undertaken to develop and evaluate the Neospora agglutination test (NAT) for the detection and quantification of IgG antibodies to N. caninum from various animal species. Compared to the ELISA method, the NAT with a cutoff value of 1:512 gave a high index of coincidence (kappa=0.807) and no cross reactivity to Toxoplasma gondii antiserum. Hence, this NAT method, which did not require a species-specific secondary antibody and expensive tools, would be easily available for the detection of antibodies to N. caninllm of various animal species.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.457-463
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• 2016

The goal of this work was to investigate the correlations between MyHC mRNA transcription and their corresponding protein expressions in porcine longissimus muscle (LM) during postnatal growth of pigs. Five DLY ($Duroc{\times}Landrace{\times}Yorkshire$) crossbred pigs were selected, slaughtered and sampled at postnatal 7, 30, 60, 120, and 180 days, respectively. Each muscle was subjected to quantity MyHCs protein contents through an indirect enzyme-linked immunosorbent assay (ELISA), to quantity myosin heavy-chains (MyHCs) mRNA abundances using real-time polymerase chain reaction. We calculated the proportion (%) of each MyHC to total of four MyHC for two levels, respectively. Moreover, the activities of several key energy metabolism enzymes were determined in LM. The result showed that mRNA transcription and protein expression of MyHC I, IIa, IIx and IIb in LM all presented some obvious changes with postnatal aging of pigs, especially at the early stage after birth, and their mRNA transcriptions were easy to be influenced than their protein expressions. The relative proportion of each MyHC mRNA was significantly positively related to that of its corresponding protein (p<0.01), and MyHC I mRNA proportion was positively correlated with creatine kinase (CK), succinate dehydrogenase (SDH), malate dehydrogenase (MDH) activities (p<0.05). These data suggested that MyHC mRNA transcription can be used to reflect MyHC expression, metabolism property and adaptive plasticity of porcine skeletal muscles, and MyHC mRNA composition could be a molecular index reflecting muscle fiber type characteristics.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.4
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• pp.573-583
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• 2015

B-32 is one of a panel of monoclonal anti-idiotypic antibodies to growth hormone (GH) that we developed. To characterize and identify its potential role as a novel growth hormone receptor (GHR) agonist, we determined that B-32 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assay assays. The results of fluorescence-activated cell sorting, indirect immunofluorescence and competitive receptor binding assays demonstrated that B-32 specifically binds to the GHR expressed on target cells. Next, we examined the resulting signal transduction pathways triggered by this antibody in primary porcine hepatocytes. We found that B-32 can activate the GHR and Janus kinase (2)/signal transducers and activators of transcription (JAK2/STAT5) signalling pathways. The phosphorylation kinetics of JAK2/STAT5 induced by either GH or B-32 were analysed in dose-response and time course experiments. In addition, B32 could also stimulate porcine hepatocytes to secrete insulin-like growth factors-1. Our work indicates that a monoclonal anti-idiotypic antibody to GH (B-32) can serve as a GHR agonist or GH mimic and has application potential in domestic animal (pig) production.

• The Plant Pathology Journal
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• v.29 no.2
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• pp.193-200
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• 2013

Trichoderma asperellum SKT-1 is a microbial pesticide that is very effective against various diseases. Our study was undertaken to evaluate T. asperellum SKT-1 for induction of resistance against yellow strain of Cucumber mosaic virus (CMV-Y) in Arabidopsis plants. Disease severity was rated at 2 weeks post inoculation (WPI). CMV titre in Arabidopsis leaves was determined by indirect enzyme-linked immunosorbent assay (ELISA) at 2 WPI. Our results demonstrated that among all Arabidopsis plants treated with barley grain inoculum (BGI) of SKT-1 NahG and npr1 plants showed no significant reduction in disease severity and CMV titre as compared with control plants. In contrast, disease severity and CMV titre were significantly reduced in all Arabidopsis plants treated with culture filtrate (CF) of SKT-1 as compared with control plants. RT-PCR results showed increased expression levels of SA-inducible genes, but not JA/ET-inducible genes, in leaves of BGI treated plants. Moreover, expression levels of SA- and JA/ET-inducible genes were increased in leaves of CF treated plants. In conclusion, BGI treatment induced systemic resistance against CMV through SA signaling cascade in Arabidopsis plants. While, treatment with CF of SKT-1 mediated the expression of a majority of the various pathogen related genes, which led to the increased defense mechanism against CMV infection.

• Food Science and Biotechnology
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• v.16 no.6
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• pp.1011-1017
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• 2007

Saeujeot (salted and fermented shrimp) and kimchi are traditional Korean fermented foods. Even though shrimp have often induced severe allergic reactions in sensitized individuals, few studies have investigated the allergenicity of shrimp. The aim of this study was to observe the changes of pH and allergenicity of raw shrimp (Acetes japonicus) and saeujeot in cabbage kimchi during fermentation using competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Fermentation was carried out at different temperatures (25, 15, and $5^{\circ}C$). The pH of cabbage kimchi added with raw shrimp or saeujeot slowly decreased at lower temperature ($5^{\circ}C$) at the end stage of the fermentation process. The binding ability of serum obtained from patients allergic to raw shrimp against shrimp tropomyosin and saeujeot in kimchi rapidly decreased during longer fermentation periods and higher temperature ($25^{\circ}C$). In conclusion, the allergenicity of both raw shrimp and saeujeot in kimchi decreased during fermentation but the decrease in allergenicity of saeujeot was greater than observed for raw shrimp.

• Korean Journal of Veterinary Service
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• v.31 no.1
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• pp.101-111
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• 2008

The purpose of this study was to investigate the protective effects against porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in suckling piglets by oral administration of IgY. Twenty piglets were divided into two groups with the same number: group I (treated with IgY) and group II (not treated). Group I was administerd orally with IgY for three days from one-day-old and experimentally challenged with PEDV and TGEV at four-day-old. The other was administered with saline solution and challenged with PEDV and TGEV at four-day-old. Serum antibody titers against PEDV and TGEV were examined by enzyme-linked immunosorbent assay (ELISA) and the detection of PEDV or TGEV antigen from feces and small intestines was performed by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescence (IFA). The antibody titers of the group I was higher than that of the other, and lasted at the end of experiment. In the detection tests of both virus from feces and small intestine, the rate of the group I was lower. Based on these results, oral administration of IgY may be effective to prevent the diarrhea caused by PEDV and TGEV.

• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1409-1414
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• 1998

To develop zearalenone-specific monoclonal antibodies, hybridoma cells were produced by fusion of myeloma cells $(P3{\times}63Ag\;V653)$ and spleen cells from BALB/c female mice immunized with zearalenone-oxime coupled to bovine serum albumin (BSA). After screening of antibody titer of them with a sandwich type enzyme-linked immunosorbent assay (ELISA), 5 hybridomas which could produced monoclonal antibodies with a high affinity for zearalenone were selected. The monoclonal antibody produced by Z-2-M26 hybridoma exhibited the high sensitivity to zearalenone and a little cross-reactivity to ${\alpha}-zearalenol$ (11%), but did not react with ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ and DON. In conclusion, the developed monoclonal antibody appeared to be a very promising immunoreagent for the future development of a specific and sensitive quantitative ELISA for zearalenone.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.197-208
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• 1987

Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.

• Journal of Veterinary Science
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• v.22 no.4
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• pp.45.1-45.13
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• 2021

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 10⁶ cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.