• Journal of fish pathology
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• v.11 no.1
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• pp.43-50
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• 1998

In order to determine whether the tumor-inducing iridoviruses and the mortality-associated iridoviruses from cultured marine fish in Korea belong to same type, we compared the immunological characteristics of these viruses. The electrophoretical pattern of structural proteins of the tumor-inducing was different from that of the mortality-associated iridoviruses. The antigenicity of structural proteins of these viruses were identified by Western blotting using two monoclonal antibodies against tumor-inducing iridovirus. Two monoclonal antibodies recognized a 150 kDa structural protein of tumor-inducing iridoviruses showed. However, the structural proteins of the mortality-associated iridoviruses did not react with these monoclonal antibodies. These results demonstrate that the antigenicity of the structural proteins of tumor-inducing iridovirus is different from that of mortality-associated iridovirus, indicating that these two iridoviruses belong to different types.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.848-853
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• 2001

This research was conducted to study the effects on antigenicities (allergenicity) and structural changes of gamma-irradiated hen’s egg albumin (ovalbumin, OVA) by heating. Three groups of OVA solution (2.0 mg/mL) were prepared; 1) heat treatment; 2) irradiation after heating; 3) heating after irradiation. Samples were isothermally heated and/or irradiated at the absorption dose of 10 kGy. Competitive indirect ELISA was individually formatted with egg-allergic patients IgE (P-IgE), and mouse murine monoclonal IgG (M-IgG) and rabbit polyclonal IgG (R-IgG) for evaluating bindinhg abilities of antibodies to OVA in the sample solutions. Binding abilities of antibodies to thermally denatured OVA were changed : R-IgG to the sample treated with above 6$0^{\circ}C$, M-IgG to that above 7$0^{\circ}C$, and P-IgE to that above 8$0^{\circ}C$, respectively. P-IgE did not well recognize OVA heated at 8$0^{\circ}C$ and the above. However, binding abilities of M-IgG and R-IgG highly in creased. Significant differences of binding abilities were not observed in all samples with the combination of heat treatment and irradiation, regardless the order of the treatment. Turbidity of samples in creased both by heating and by irradiation, and the increase by irradiation was much higher than by heating. These results showed that allergenicity of OVA reduced by gamma irradiation was not affected by heating.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.8
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• pp.417-424
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• 2019

Despite its superior ability to show distinct cellular morphology and for long-term storage, conventional tissue fixation by formalin has many drawback, including slower fixation, the exposure to harmful chemicals and extensive protein modification. Herein, we assessed the effects of rapid microwave-assisted tissue fixation on histological examination and on protein integrity by comparing these microwave irradiation fixated tissues with the formalin-fixed tissues. One of the paired mouse tissues (liver and kidney) was fixed in formalin and the other was fixed by using microwave irradiation in phosphate buffered saline. Each slide from the paraffin-embedded tissues was examined by H & E staining for the adequacy of fixation and by immunohistochemical staining for antigenicity in a blinded fashion. Evaluation of protein recovery and the protein quality from the fixed tissues were analyzed by the BCA method and Western blotting, respectively. The results from H & E staining and immunohistochemical staining showed that the sections obtained from microwave-fixed tissues under our experimental conditions were comparable to those of the formalin-fixed tissues except for the integrity of RBCs. Furthermore, proteins were effectively extracted from the microwave-fixed tissues with acceptable preservation of the proteins' quality. Taken together, this microwave-assisted tissue processing yields a quick fixation and better protein recovery in higher amounts, as well as the adequacy of fixation and the antigenicity being comparable to formalin-fixed tissues, and this all suggests that this new fixation technique can be applied in an environment where rapid tissue fixation is required.

• Journal of Veterinary Science
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• v.23 no.4
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• pp.55.1-55.12
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• 2022

Background: ASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection. Objective: To identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays. Method: We used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide. Results: The results of our prediction revealed that the possible antigen epitope regions were A23-29, A36-45, A72-94, A114-120, A124-130, and A137-150. The indirect ELISA showed that the peptides A23-29, A36-45, A72-94, A114-120, and A137-150 have good antigenicity. Moreover, the A36-45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44. Conclusions: Our study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.

• Parasites, Hosts and Diseases
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• v.54 no.2
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• pp.239-241
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• 2016

Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

• Journal of Korean Dental Science
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• v.9 no.2
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• pp.55-62
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• 2016

Purpose: Alveolar bone develops with tooth eruption and is absorbed following tooth extraction. Various ridge preservation techniques have sought to prevent ridge atrophy, with no superior technique evident. Collagen has a long history as a biocompatible material. Its usefulness and safety have been amply verified. The related compound, atelocollagen, is also safe and displays reduced antigenicity since telopeptides are not present. Materials and Methods: The current study evaluated whether the $Rapiderm^{(R)}$ atelocollagen plug (Dalim Tissen, Seoul, Korea) improves tissue healing of extraction sockets and assessed the sequential pattern of bone regeneration using histology and microcomputed tomography in six beagle dogs. To assess the change of extraction socket, hard tissues were examined 2, 4, 6, and 8 weeks after tooth extraction. Result: The experimental groups showed better bone fill with slow remodeling process compared to the control groups although there was no statistical difference between groups. Conclusion: The atelocollagen seems to have a tendency to slow bone remodeling in the early phase of healing period and maintain remodeling capacity until late phase of remodeling. Also, use of atelocollagen increased the bone-to-tissue ratio compared to healing of untreated extraction socket.

• Journal of Environmental Health Sciences
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• v.28 no.5
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• pp.28-34
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• 2002

Antigenicity of Rhodotrula rubra isolated from pulmonary tissue of pulmonary tuberculosis patients was studied by means of agglutination reaction with R. rubra whole cell antiserum. And the serological reactivity of crude polyfac charide from R. frubra, Candida albicans, Candida tropicalis, Candida, glabrata, and Saccharomyces cerevisiae ATCC 26603 with antiserum to R. rubra whole cell was studied by means of immunodiffusion test. R. rubra showed stationary phase after 48h when it was cultured in GYEP broth. While agglutinogen titer was 1:64 at lag phase, agglutinogen titer was 1 :256 after 20h. After growth of R. rubra on different 11 media, nutritional environment showed similar agglu-tination reartivity. The agglutinogen titer of C. albicans, C. tropicalis, C. giabrata, which were isolated from patient's expectoration, to R. rubra antiserum by means of agglutination reaction were 1:16, respectively. But, Sacch. cervisiae ATCC26603 was negative. Those results were lower than that of R. rubra agglutinogen titer 1:256. As a result of immu-nodiffusion test with crude polysaccharide extracted from cell wall of R. rubra, C. albicans, C. tropicalis, C. glabrata, Sacch. cervisiae ATCC26603, precipitin line was found only with R. rubra, of which antibody titer was 8.

• BMB Reports
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• v.28 no.4
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• pp.353-358
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• 1995

Epitope tagging is the process of fusing a set of amino acid residues that are recognized as an antigenic determinant to a protein of interest. Tagging a protein with an epitope facilitates various immunochemical analyses of the tagged protein with a specific monoclonal antibody. The monoclonal antibody H8 has subtype specificity for an epitope derived from the preS2 region of hepatitis B virus surface antigen. Previous studies on serial deletions of the preS2 region indicated that the preS2 epitope was located in amino acid residues 130~142. To test whether the amino acid sequence in this interval is sufficient to confer on proteins the antigenicity recognizable by the antibody H8, the set of amino acid residues in the interval was tagged to the amino terminal of ${\beta}$-galactosidase and to the carboxyl terminal of the truncated $p56^{lck}$ fragment. The tagged ${\beta}$-galactosidase, expressed in Escherichia coli, maintained the enzymatic activity and was immunoprecipitated efficiently with H8. The tagged $p56^{lck}$ fragment, synthesized in an in vitro translation system, was also immunoprecipitated specifically with H8. These results demonstrate that the amino acid sequence of the preS2 region can be used efficiently for the epitope tagging approach.

• Biomolecules & Therapeutics
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• v.6 no.1
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• pp.50-55
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• 1998

Antigenic potential of a recombinant human erythropoietin (rhEPO) produced by Dong-A charm. Co. Ltd. was examined by active systemic anaphylaxis (ASA) test in guinea pigs, mouse-rat passive cutaneous anaphylaxis (PCA) reaction and passive hemagglutination (PHA) test. In ASA test, rhEPO induced the signs of restlessness, rubbing or licking nose, sneezing and coughing in the animals immunized with rhEPO 1000 lU/kg alone or rhEPO 1000 lU/kg incorporated into Freund\\\\`s complete adjuvant. In the mouse-rat PCA test, only one of six sera from the animals immunized with rhEPO 1000 lUng incorporated into Alum showed positive result. In the PHA test, rhEPO revealed negative results in all of the rhEPO-immunized groups. From these results, rhEPO was considered to produce IgE in guinea pigs and mice, but not IgG and/or IsM in mice. The results of this study were similar to those of the other recombinant human erythropoietin and these positive results were thought to be caused due to the fact that rhEPO were heterogeneous proteins to guinea pigs and mice. Considering the fact that rhErO has an identical structure with indigenous human erythropoietin, rhEPO is not thought to cause immunological problems in clinical use.

• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.565-570
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• 2001

The diagnosis of swine sarcoptic mange (Sarcoptes scabiei var. suis) was investigated by ELISA in order to replace current diagnostic methods such as skin scraping, scratching index, or lesion score of dermatitis. The current methods need many efforts and much times and cost much. They can not handle many samples simultaneously. Therefore, in this research we developed ELISA that can handle many samples at a time. The antigens of swine sarcoptic mite were isolated and examined by 12.5% SDS-PAGE and silver staining. The antigenicity of antigen was confirmed by Western blotting using the swine from the artificailly-infested swine with swine sarcoptic mite. The optical density (OD) values of the artificailly-infested positive sera and the naturally-infested positive sera of sows were measured and read in order to confirm the stability of antigens, the reproducibility and validity (sensitivity and specificity) of the manufactured ELISA of swine sarcoptic antigens. In above results, the developed ELISA would be possible to use the diagnostic tool of sarcoptic mange if OD values of piglets, fattening pigs and sows are interpreted reasonably and classified as mange-free and infested.