• Journal of the Korean Society of Visualization
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• v.20 no.3
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• pp.136-145
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• 2022

Later flow immunoassay (LFIA) is a protein analytical method based on immunoreaction. On the LFIA based protein analytical method, bioreceptor molecule plays a key role, and so a system that evaluates and manages the binding affinity of bioreceptor is needed to secure detection reliability. In this study, Lateral Flow Immunoassay based rapid Bioreceptor Screening Method (rBSM) is presented that provide a simple and quick evaluating method for the binding affinity to the target protein of the antibody as model bioreceptor. To verify this evaluation method, Virus-like particles (VLP) and anti-VLP antibodies are selected as a model norovirus, which is target protein, and the candidate bioreceptors respectively. Among the 5 different candidate antibodies, appropriate antibody could be sorted out within 30 minutes through rBSM. In addition, selected antibodies were applied to two representative LFIA based techniques, sandwich assay and competitive assay. Among these methods, sandwich assay showed more effective VLP detection method. Through applying selected antibodies and techniques to the commercialized mass production lines, an VLP detecting LFIA kit was developed with a detection limit of 10¹² copies/g of VLPs in real samples. Since this proposed method in this study could be easily transformable into other combinations with bioreceptors, it is expected that this technique would be applied to LFIA kit development system and bioreceptor quality management.

• Parasites, Hosts and Diseases
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• v.33 no.3
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• pp.201-210
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• 1995

This study aims to assess the possible strain-dependent variations in detection of ToxopLosmn antigens and antibodies. The virulent RH strain or avirulent Beverley strain of T gondii were injected into mice, intraperitoneally, and their antigens, antibodies and parasites were identified from the blood or tissues: liver, brain and spleen by ELISA, Western blot and PCR. In mice infected with RH strain, circulating antigens and parasitemia were first detected from 2 days after infection, and ToxopIasma DNA were found in the blood, liver, brain and spleen from 3 days after infection. It was impossible to detect specific IgM and IgG antibodies to T gondij and any specific band was not found by Western blot. In mice infected with Beverley strain, circulating antigens were detected between day 10 and day 35. The Toxoplusma DNA was found in the blood and liver from day 15 until day 60, and in the brain from day 20. But Toxoplosma DNA in the spleen were mainly detected between day 10 and day 30. The IgM antibodies were first appeared on day 10 post-infection, and were noted obviously increased between day 15 and 25. The IgG antibodies were first detected on day 15, and showed progressively increased titers. The antibody binding bands were specific according to infection period. Sera from mice infected with Beverley strain reacted mainly with the antigen of 27.5-kDa and 32.5-kDa. In conclusion, mice infected with RH strain revealed Toxoplosma antigens strongly, but not antibodies. However. mice infected with Beverley strain revealed both the Toxoplasma antigens and antibodies. The present results showed that immune responses are different between avirulent and virulent T gonnii.

• Korean Journal of Veterinary Service
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• v.22 no.4
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• pp.371-375
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• 1999

Total 1,434 sera collected from 72 pig farms were tested for the detection of porcine reproductive and respiratory syndrome (PRRS) virus antibodies. The overall seroprevalence of PRRS virus antibodies was 49.3% (707/727). Of 72 farms tested 59 (81.9%) farms had at least one or more than one pigs with PRRS virus antibodies. The seroprevalence of PRRS virus antibody varied with age. Seroprevalence of PRRS virus antibody in 1 to 30-day-old, 31 to 40-day-old, 41 to 50-day-old, 51 to 60-day-old, and over 61-day-old pig were 27.4%, 52.3%, 57.9%, 52.7%, and 68.2%, respectively. Gilt showed relatively higher seroprevalence (61.2%) than sow (29.2%) and boar (38.3%). In most farms, the infection of PRRS virus was chronic and confined to grower or finisher. This pattern of infection suggests that partial depopulation of the infected herds appears be one of the measures to eradicate the PRRS virus infection. High seroprevalence of the PRRS virus antibody in gilts and boars indicates that the infected gilts and boars in the breeding farms are the major source of the PRRS virus infection, and also play an important role in spreading the PRRS virus between fan mates or herds.

• Clinical and Experimental Reproductive Medicine
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• v.12 no.1
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• pp.41-46
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• 1985

Various immunoserologic and cellular immunity techniques have been used to explore the presence of antisperm antibodies in the serum and seminal plasma of male patients and in the blood and genital fluid of infertile women. Several recent comparative investigations using various assays to detect and quantitate levels of antibody to human spermatozoa have produced widely varying results. So the first WHO workshop on iso- and autonatibodies to human spermatozoa in 1974 tried to establish some unification in the techniques used. The purpose of this study is to compare the results of two methods-the Kibrick macro-agglutination test and the Isojima micro-immobilization test-using the same test materials based on recommandation from WHO workshop. The results are as follows: 1. Twenty normal controls showed negative reactions in all the 2 tests. Out of 25 patients, the positive sera were noted in 15 (60%) on the Kibrick test and 13 (51%) on the Isojima test. 2. Twelve (48%) out of 25 patients showed positive reactions in the two tests, and 16 (64%) out of 25 patients showed positive reaction in one or more tests. 3. The titers of the antisperm antibodies on the Kibrick test was higher than that on the Isojima test. Therefore, it seems to be possible to increase the chances of detection of the antisperm antibodies, if two tests are imployed.

• The Journal of the Korean Society for Microbiology
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• v.20 no.1
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• pp.103-107
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• 1985

Female ICR mice were vaccinated against Toxoplasma gondii(RH strain), infected 2 weeks later and after recovery mated to normal ICR males. Control matings were with normal ICR females. The progeny of the above matings were weaned at 1 week, vaccinated at 1, 2, 3, 4 or 5 weeks of age and infected 2 weeks later with lethal Toxoplasma tachyzoites. As assessed by survival, the effectiveness of vaccination among offspring of vaccinated-recovered mothers was greatly impaired than that of control mice, with respective of age : where mice did survive, recovery was greatly delayed relative to the controls. The protective effect of vaccination among infants born to control mothers was also blocked by maternal specific antibodies, by administration of high-titered specific antibodies or by transfer of nylon wool adherent immune-spleen cells, but was augmented by transfer of nylon wool passed immune-spleen cells. These results indicate that this impairment of vaccination may be due to the transmission of maternal specific antibodies to the offspring which acts to suppress both priming by the vaccine and the generation of parasite-specific helper T cells.

• Korean Journal of Veterinary Service
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• v.43 no.3
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• pp.173-179
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• 2020

The objective of this study was to evaluate the usefulness of detection of PRRSV and PRRSV-specific antibodies in oral fluids for monitoring of PRRSV infection in endemic farms. The level of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in five age groups of pigs (6, 9, 12, 16 weeks of age and gilts). The samples (25 serums and 5 oral fluids/per a farm) were collected from 5 different farms endemically infected by PRRSV. Both serum and oral fluid samples were tested for PRRSV by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and for anti-PRRSV antibodies by two commercial PRRSV ELISA kits. ELISA mean s/p ratios (2.98 vs 1.63) and positive rate (84.0% vs 68.8%) of the oral fluid samples showed significantly higher levels but had similar patterns to the seroprofile of the blood samples. The PRRSV positive rate of oral fluid and serum samples was 40.0% and 44.0% respectively. In conclusion, the use of oral fluids for PRRS monitoring in endemic farms is strongly recommended.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.395-403
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• 1996

Monoclonal antibodies(MAbs) against field isolates of the bovine rotavirus A strain(G6), V strain(G10) and reference I-801 strain(G8) were produced and characterized. Six MAbs(4C2, 4D9, 5E1, 5E7, 5D5, 3E4) against A strain had neutralizing activity and reacted only with the G6 bovine rotaviruses determined by fluorescence focus neutralization (FFN) test. Otherwise, five neutralizing MAbs(1G2, 2G6, 5E2, 5E12, 5H7) against I-801 strain neutralized the G6 and G8 bovine rotaviruses. Five non-neutralizing MAbs(5F12, 7F12, 5E11, 2A11, 2B12) were VP6-specific and cross-reacted with all bovine and porcine rotaviruses examined by fluorescence antibody(FA) test. None of the MAbs reacted with bovie viral diarrhea virus(BVDV) and bovine coronavirus(BCV) determined by FA and FFN test.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.337-348
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• 1996

The proportion of leukocyte subpopulation in the host is of a great importance in understanding their functions and disease progress. Many methods have been developed to seperate leukocytes and to measure their activities. Characterization of immune cell subpopulations in Korean native cattle was performed using a set of monoclonal antibodies specific which are specific to bovine leukocyte differentiation antigen. Peripheral blood leukocytes from fifty Korean native and ten Holstein cattle were collected and analyzed for the investigation of leukocyte subpopulation by using monoclonal antibodies and flow cytometry. The result indicated that Korean native cattle have significantly higher proportion of leukocyte subpopulations expressing MHC class II molecules and BoCD4 than Holstein cattle.

• BMB Reports
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• v.33 no.2
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• pp.188-192
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• 2000

Recombinant protein G has been utilized in the purification of antibodies from various mammalian species based on the interaction of antibodies with protein G. The interaction between immunoglobulin and protein G may not be restricted to the Fc protion of antibodies, as many different $F(ab)_2$ or Fab fragments can also bind to protein G. I found both FAb $F(ab)_2$ of 145-2C11, a hamster anti-mouse $CD3{\varepsilon}$ antibody, bound to the protein G-sepharose. Interestingly, Fab and $F(ab)_2$ of 145-2C11 did not bind to the protein A-sepharose. The binding of Fab and $F(ab)_2$ of 145-2C11 to protein G provided a useful method to remove proteases, chopped fragments of the Fc region, and other contaminating proteins. The remaining intact antibody in the protease reaction mixture can be removed by using a protein A-sepharose, because the Fab and $F(ab)_2$ portions of 145-2C11 did not bind to protein A-sepharose. The specific binding of Fab and $F(ab)_2$ portions of 145-sC11 to a protein G-sepharose (though not to a protein A-sepharose) and binding of intact 145-2C11 to both protein A- and G-sepharose will be useful in developing an effective purification protocol for Fab and $F(ab)_2$ portions of 145-2C11.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.2
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• pp.534-537
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• 2004

We produced twelve monoclonal antibodies(mAbs) against thyroglobulin and characterized the bindig profiles. Among them, three mAbs(TN-1, TN-2 and TN-3) were further characterized their binding specificities. TN-2 had a potent lupus anticoagulant activity and potentiated the anticoagulant effect of venom phospholipase A₂. he anticoagulant mechanism of TN-2 was elongation of the partial thromboplastin time and binding to phosphatidylserine which may have a pivot role in blood coagulation. And TN-2 was cross-reacted with ss-DNA and ds-DNA and had a characteristic of autoantibody. These results suggest that TN-2 may provide a useful tool for studying the correlation between autoimmune thyroiditis and its therapeutic effect.