• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.879-890
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• 2007

The identification of tumor antigens is essential for the development of anticancer therapeutic vaccines and clinical diagnosis of cancer. SEREX (serological analysis of recombinant cDNA expression libraries) has been used to identify such tumor antigens by screening sera of patients with cDNA expression libraries. SEREX-defined antigens provide markers for the diagnosis of cancers. Potential diagnostic values of these SEREX-defined antigens have been evaluated. SEREX is also a powerful method for the development of anticancer therapeutics. The development of anticancer vaccines requires that tumor antigens can elicit antigen-specific antibodies or T lymphocytes. More than 2,000 antigens have been discovered by SEFEX. Peptides derived from some of these antigens have been evaluated in clinical trials. This review provides information on the application of SEREX for identification of tumor-associated antigens (TAA) for the development of cancer diagnostics and anticancer therapeutics.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.993-1001
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• 2007

The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.301-310
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• 1999

Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases. The bacterium produces several virulence factors. Of the factors, protective antigen (PA) of tripatite toxin has been identified as a central component in the pathogenesis of anthrax. However, precise roles of PA and other cellular components in the reaction with the target cells remain to be elucidated, especially in the initial stage of the disease. Three B anthracis antigens were prepared for investigation; PA, sonicated cellular antigens (S-Ag) and formalin-inactivaed whole cell antigens (W-Ag). PA was purified from culture supernatant of the bacterium using FPLC system with MonoQ. S-Ag and W-Ag were prepared by sonication and formalin inactivation of the cultured cells, respectively. Purity of the antigens was confirmed by SDS-PAGE and Western blot analysis. The roles of these antigens in the production of inflammatory mediators such as NO, IL-6 and $TNF{\alpha}$ from mouse peritoneal macrophages were investigated. PA alone did not induce the production of the inflammatory mediators while the other antigens, S-Ag and W-Ag, did in a dose and time dependent manner. These results suggested that in addition to major virulence factors, other cellular antigens are also involved in the initial stage of the disease by the induction of inflammatory mediators.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.841-846
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• 2007

The identification of tumor antigens is essential for the development of anticancer therapeutic vaccines and clinical diagnosis of cancer. SEREX (serological analysis of recombinant cDNA expression library)has been used to identify such tumor antigens by screening sera of cancer patients with cDNA ex-pression libraries. SEREX-defined antigens provide markers for the diagnosis of cancers. SEREX is also a powerful method for the development of anticancer therapeutics. The development of anticancer vaccines requires that tumor antigens can elicit antigen-specific antibodies or T lymphocytes. This re-view provides information on the application of SEREX for discovery of tumor antigens.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.73-85
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• 1983

The advantages of the enzyme-linked immunosorbent assay(ELISA) are its sensitivity and its simplicity in detecting IgM and IgG antibodies. For applying the ELISA to the diagnosis of typhoid fever, first of all, experiments were performed to determine which concentration of killed whole cell antigens and lipopolysaccharide(LPS) antigens of S. typhi(0.901 w) were optimally coated to the wells of the polystyrene and polyvinylchloride microplate, using the hyperimmune sera from rabbits against S. typhi. By using both kinds of antigens of S. typhi adsorbed to the ELISA microplate, the changing patterns of IgG and IgM antibodies in the sera from rabbits responding to the killed whole cell antigens of S. typhi(0901 w) during the prolonged immunization were serially traced by the ELISA. At the same time, the level of antibodies against S. typhi in sera fron patients with typhoid fever and from normal healthy persons were measured by the ELISA employing the killed whole cell antigens and LPS antigens as the coating antigens. The results obtained were summerized as follow: 1. The optimal concentration of the killed whole cell antigens, which were more easily adsorbed to the polystyrene plate than the polyvinylchloride plate, was $10^8cells/ml$ of carbonate buffer(pH. 9.6) on the wells of the polystyrene plate when treated at $37^{\circ}C$ for 4 hours. On the other hand, the optimal concentration of lipopolysaccharide antigens, which were adsorbed only to the polyvinylchloride plate, was $100{\mu}g/ml$ of carbonate buffer(pH. 9.6) on the wells of the polyvinylchloride plate when treated at $37^{\circ}C$ for 4 hours. 2. IgM antibody response were dominating in rabbits responding to the killed whole cell antigens of S. typhi(0.901 w), and were more specific to the LPS antigens than to the killed whole cell antigens in the ELISA. Good correlations were made between the IgM titers by the ELISA and the aggglutinating titers of sera from the immunized rabbits. 3. Both IgG and IgM agglutination titers by the ELISA in sera from most of patients with typhoid fever were above 1:320 but those in sera from most of normal, healthy persons were below 1:80. 4. There were close correlations between the antibody titers by the ELISA and the agglutinating titers to the killed whole cell antigens in the tested human sera, IgM titers being more correlated with the agglutinating titers than IgG titers. But a little correlations were made between the antibody titers by the ELISA and the agglutinating titers to the LPS antigens. 5. IgM titers in the tested human sera were similar to IgG titers detected by the ELISA employing the killd whole cells antigens and the LPS antigens. 6. Good correlations were made between the antibody titers demonstrated by the ELISA performed on the killed whole cell antigens and the LPS antigens as the different, coating antigens on the ELISA microplates.

• Annals of Laboratory Medicine
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• v.38 no.6
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• pp.599-603
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• 2018

Frequencies of red blood cell (RBC) blood group antigens differ by ethnicity. Since the number of immigrants is increasing in Korea, RBC antigens should be assessed in children/youths with parents of different ethnicities to ensure safe transfusions. We investigated the frequency of RBC antigens, except for ABO and RhD, in 382 children and youths with parents having Korean and non-Korean ethnicities. Subjects were divided into those with ethnically Korean parents (Korean group; N=252) and those with at least one parent of non-Korean ethnicity (non-Korean group; N=130). The 37 RBC antigens were genotyped using the ID CORE XT system (Progenika Biopharma-Grifols, Bizkaia, Spain). The frequencies of the Rh (E, C, e, $hr^S$, and $hr^B$), Duffy ($Fy^a$), MNS ($Mi^a$), and Cartwright ($Yt^b$) antigens differed significantly between the two groups. Eight and 11 subjects in the Korean and non-Korean groups, respectively, exhibited negative expression of high-frequency antigens, whereas 14 subjects in the non-Korean group showed positive expression of low-frequency antigens. The frequency of RBC antigens has altered alongside demographic changes in Korea and might lead to changes in distribution of RBC antibodies that cause acute or delayed hemolytic transfusion reaction.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.62-66
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• 2001

This study was performed to investigate antigenic expression patterns in the course of HSV-1 infection. In SDS-PAGE analysis, HSV-1 antigens were detected, and among them, antigens in the size of 39, 47, 63, 86, 101, 105, 135, 159, and 181 kDa appear to be expressed in the most dominant forms. BALB/c mice were infected with HSV-1 for 29 days and antigenic expression from HSV-1 was investigated by Western blot analysis using anti-HSV-1 sera collected every two days from BALB/c mice infected with HSV-1. Most of HSV-1 antigens appeared sporadically as the infection progressed. However, antigens in the sizes of 63kDa and 135kDa were expressed from day 1 and 3, respectively, and existed continuously during the course of infection for 29 days, suggesting that they are the most dominant antigens inducing immune response durign HSV-1 infection, and they could be the target antigens for the development of vaccines. The isotype levels of IgA, IgGl, and IgM increased till the 17 th day infection and then started to decrease. During this course. IgGl was the most dominant isotype. In an indirect immunofluorescent assay, antibodies exhibited surface binding to the Vero cell infected with HSV-1, demonstrating that HSV-1 antigens are expressed on the surface of Vero cells.

• Korean Journal of Veterinary Service
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• v.25 no.3
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• pp.295-297
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• 2002

This experiment was done to set up the immunohistochemical detection method for infectious hematopoietic necrosis virus(IHNV) antigens in the monolayers of CHSE-214 cell cultures inoculated with IHNV. Specific identification of IHNV antigens was detected in the cytoplasms of infected cells by the use of monoclonal antibodies to glycoproteins. The specific positive signal was observed as a distinct red color. The result showed that streptavidin alkaline phosphatase immunohistochemistry specifically identified IHNV antigens in infected cultured cells.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.171-176
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• 1986

In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay (ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: 1. The specificity (95% confidence interval in negative sera of bovine fascioliasis; Mean+$2{\times}SD$ of absorbence) of the first (MW>150,000) and the second antigens(MW 120,000) were 93.7%, but those of others including crude antigen showed 100%. 2. The sensitivity(positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6%, 87.5% and 0%, respectively, but those of others showed all 100%. 3. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8. 43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. 4. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the fourth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. 5. The wheal size for bovine infected with F. hepatica was $2.46{\pm}0.15cm$ in the intradermal test antigen(saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that. of intradermal test antigen, whereas those of the fouth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.105-111
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• 2005

Canine sarcoptic mite (Sarcoptes scabiei var. canis) is ectoparasite which burrow usually in the stratum corneum of the skin of dogs. Antigens from the burrowing mites induce humoral and cellmediated immune responses in the hosts. The effect of antigenecity induced by somatic antigens of house dust mite (Dermatophagoides pteronyssinus) isolated by continuous elution has been evaluated in canine sarcoptic mites infestation. Continuous elution was carried out in 7.5% SDS-PAGE to isolate proteins of common antigens from somatic antigens of house dust mite. These eluted proteins from somatic antigens of house dust mite were confirmed by Western blotting in 7.5% SDS-PAGE, and eluted proteins (65, 60 kDa) were isolated. To evaluate the antigenetic effect of eluted proteins, eight dogs were divided as 4 groups such as non-vaccinated and non-challenged control (Group I), challenged control (Group II), vaccinated (Group III), and vaccinatedandchallenged (Group IV) groups. Group II and IV were artificially infested canine sarcoptic mites. Group III and IV were immunized with eluted proteins (65, 60 kDa). At the 6th week of the vaccination, the antibody titers of Group of IV were statistically significant higher than those of Group II (p<0.05). And antibody titers of Group III were also statistically significant higher than those of Group I (p<0.05). From these result, it is possible to replace somatic antigens of canine sarcoptic mites with eluted proteins from somatic antigens of house dust mites in order to diagnose and prevent the canine sarcoptic mite infestations.