• Title/Summary/Keyword: Sera

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Optimized Serological Isolation of Lung-Cancer-associated Antigens from a Yeast Surface-expressed cDNA Library

  • Kim, Min-Soo;Choi, Hye-Young;Choi, Yong-Soo;Kim, Jhin-Gook;Kim, Yong-Sung
    • 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.

Evaluation of enzyme-linked immunosorbent assay (ELISA) for detection of olive flounder antibodies to viral hemorrhagic septicemia virus (VHSV, genotype IVa) using two Novirhabdovirus antigens

  • Min-Seok Jang;Myung-Joo Oh;Wi-Sik Kim
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.9-15
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    • 2024
  • An enzyme-linked immunosorbent assay (ELISA) with two Novirhabdovirus antigens (viral hemorrhagic septicemia virus, VHSV and infectious hematopoietic necrosis virus, IHNV) was used to detect specific antibodies against VHSV from olive flounder (Paralichthys olivaceus) sera. In ELISA plates with VHSV culture supernatants (VHSV-Ag plate), optical density (OD) values for sera from olive flounder with VHS history (VHS sera) ranged from 0.64±0.36, and those of sera from fish without VHS history (non-VHS sera) ranged from 0.26±0.26. In IHNV-Ag plate, the OD values (0.43±0.28) for VHS sera were quite low compared to those in VHSV-Ag plates, while the OD values for non-VHS sera were almost similar. When the OD values for each serum were calculated by subtracting the OD values in the IHNV-Ag plate from those in the VHSV-Ag plate, the corrected OD values were significantly different between VHS sera and non-VHS sera. The results were completely in line with fish histories of VHS epizootics. It was considered that the corrected OD values may represent the true values recognized by VHSV-specific antibodies.

Distribution of Antibodies Against Hepatitis B virus and Hepatitis C virus in Human Sera (사람 혈청내 B형과 C형 간염바이러스에 대한 항체분포)

  • Choi, Jin-Sub;Seong, In-Wha
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.129-135
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    • 1997
  • Serum samples from 123 males and 123 females collected by age in 1996 were analyzed for antibodies against surface antigen of Hepatitis B virus and C22-3, C200 antigens of Hepatitis C virus. Sera from the children under the age of 10 showed 30% seropositivity to the surface antigen of Hepatitis B virus, 33.3% in $10{\sim}19$ year group, 20% in $20{\sim}29$ year group, 17.6% in $30{\sim}39$ year group, 3.3% in $40{\sim}49$ year group, 5.9% in $50{\sim}59$ year group, 8,3% in $60{\sim}69$ year group, 2.9% in $70{\sim}79$ year group, but antibody could not found in $80{\sim}86$ year group. 12 out of 123 male sera were positive, 19 out of 123 female sera were positive and overall rate of positivity of antibody against surface antigen of Hepatitis B virus was 12.6%. Serum samples from peoples under the age of 30 had not antibody against C22-3, C200 antigens of Hepatitis C virus. The positivity rate was 2.9% in $30{\sim}39$ year group. 5 out of 30 sera from $40{\sim}49$ year age group were positive, and 3 positive sera showed extremely high titer (1:524,288) but the titers of two remaining sera were 1:32, 1:8,192 respectively. 5.9% was positive in $50{\sim}59$ year group, 8.3% in $60{\sim}69$ year group, 11.8% in $70{\sim}79$ year group but all negative in $80{\sim}86$ year group 6 out of 123 male sera were positive (4.9%), 9 out of 123 female sera were positive (7.3%). Overall rate of positivity of antibody against C22-3, C200 antigen of Hepatitis C virus was 6.1 %. None out of 246 sera had both antibodies against Hepatitis B virus and Hepatitis C virus.

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Serosurvey for antibodies against brucellosis in pigs (돼지 brucellosis에 대한 항체가 조사)

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.153-157
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    • 2011
  • In order to investigate serum antibodies for detection of brucellosis in pigs, a total of 1208 sera were tested by Rose Bengal test (RBT), the standard tube agglutination test (STAT) and competitive ELISA (cELISA). The sera were collected from pigs of Gyeonggi, Chungnam, Chungbuk, Jeonnam and Jeonbuk, provinces during the period 2002 to 2004. All the sera were screened by RBT, and were confirmed by STAT and cELISA. Among 1208 sera, 26 sera (2.2%) were positive in screening test. All the 26 positive sera were positive by STAT, while all the sera were negative by cELISA. On the basis of this study, farmed pigs may be exposed to Brucella species. Furthermore, these results suggest that establishment of diagnoses for detection of porcine brucellosis is necessary.

Effects of Storing Time on the Values of the Clinical Biochemistry in Sprague-Dawley(SD) Rats (랫드 혈청의 저장기간에 따른 혈액생화학치 변화)

  • Son, Hwa-Young;Lee, Hyun-Sook;Kim, Young-Hee;Kim, Yong-Beom;Kim, Il-Hwan;Ha, Chang-Su;Kang, Boo-Hyon
    • Korean Journal of Veterinary Pathology
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    • v.3 no.2
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    • pp.87-91
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    • 1999
  • The present study was undertaken to compare the variation on serum biochemical values by storage in the rats. Sera were prepared from 30 Sprague-Dawley rats of each sex. 5 aliquots from each serum were placed in a -80$^{\circ}C$ freezer with the exception of I aliquots which was analyzed immediately. The analysis was performed on the following months; 1, 2, 3, 6, and 12 months after freezing. The parameters measured) were aspartate aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(ALP), blood urea nitrogen(BUN) creatinine(CRE), glucose(GLU), total cholesterol(TCHO), triglyceride (TG), total protein(TP), albumin(ALB), total bilirubin(TBIL), calcium(Ca$\^$++/), inorganic phosphorus(IP), creatine kinase (CK), phospholipid(PL), albumin-globulin ratio(A/G), sodium(Na$\^$+/), potassium(K$\^$+/), and chloride(Cl$\^$-/) The statistical analysis with Repeated Measures ANOVA, did not show statistical significance in the parameters of AST, ALT, BUN, TG, CK, A/G, Na$\^$+/ of 1 month freezed sera, in those of AST, TG, CK, K$\^$+/) of 2 month freezed sera, in those of AST, ALT, BUN, CRE, TCHO, TP, TBIL, CK, PL, Na$\^$+/), K$\^$+/), Ct on month fteezed sera, in those of Cl$\^$-/ of 6 month fteezed sera, and in those of ALT, TG, ALB of 12 month freezed sera in male SD rats. On the other hand, it did not show statistical significance in the parameters of AST, ALT, ALP, BUN, GLU, TCHO, TG, TBIL, CK, PL, A/G, Na$\^$+/ of 1 month freezed sera, in those of AST, TCHO of 2 month freezed sera, in those of AST, BUN, CRE, TCHO, TP, TBIL, CK, PL of 3 month freezed sera, in those of TCHO, IP, PL of 6 month freezed sera, and in those of ALB of 12 month freezed sera in female SD rats. On the basis of the results, although there are some statistical variations in the biochemical values of the sera, it is suggested that if sera are analysed at the same time before 12 months storage in a -80 $^{\circ}C$ freezer, the storing time does not affect the biochemical evaluation of the sera in SD rats.

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Immunological Characterization of Antigen from Cysticercus and Sparganum, and their Application to the Development of Immunodiagnostic System 2. Immunological Characteriaation of Crude Antigenic Components from Sparganum (Cysticercus와 Sparganum에서 추출한 조항원의 면역학적 특성과 그의 면연진단에 응용 2. Sparganum의 조항원성 분의 면역학적 특성)

  • ;James Yang
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.429-440
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    • 1989
  • Spirometru erinocei의 유충인 sparganum에서 추출한 조항원 단백질을 항원으로 하여 sparganosis, cysticercosis, hydatidosis환자의 IgG 항체와 정상인의 IgG 항체를 혈청반응시켜 ELISA와 EITB에 의해 교차반응을 일으키는 비 특이항원 성분과 종특이항원 성분을 추구하였다. 1. sparganum에서 0.01 M PBS (PH 7.4)로 추출한 조항원 단백질을 SDS-PAGE로 전개하여 290 Kd에서 23 Kd범위의 분자량을 가진 25개의 단백질이 분획되었다. 2. sparganum의 조항원을 항원으로하여 ELISA로 sparganosis, cysticercosis, hydatidosis 환자의 190 항체와의 혈청 반응치는 sparganosis환자에게서는 0.44 $\pm$ 0.07에서 1.90 $\pm$ 0.03으로 negative control normal sera의 0.D (0.15 $\pm$ 0.03)를 기준으로 하였을 때 모두 양성이며 민감도(sensitivity)가 100 %이었으며 cysticercosis, hydatidosis환자혈청에서 양성반응이 나타났으며 교차반응도 있었다. 3. EITB에서는 spargancsis환자의 IgG항체에 의해 16개의 항원성분이 인지되었으며 이 중 6개의 항원성분이 정상인의 혈청에서도 인지되어 교차반응을 일으키는 항원성분이었으며 cysticercosis환자혈청에서 인지된 4개의 항원성분 중 2개의 항원성분이 sparganosis 환자혈청에서 인지된 것과 같았으며 hydatidosis환자의 190항체에 의해 인지된 19개의 항원성분 중 12개의 항원성분이 sparganosis환자혈청에서 인지된 항원 성분과 같았다. 4. 290 Kd, 200 Kd, 28 Kd의 항원성분은 sparganosis환자의 196항체에서만 인지되었고 228 Kd, 152 Kd, 66 Kd항원성분은 hydatidosis환자의 19G항체에서만 인지되었으며 66 Kd항원성분은 sparganosis, cysticercosis, hydatidosis, 정상인의 혈청에서 모두 인지되었다. We studied the serological reaction between the specific and nonspecific antigenic components from metacestode (plerocercoid) of spiromeko erimacei and IgG antibodies in sparganosis, cysticercosis. hydatidosis patients and normal human sera by ELISA and EITB. We prepared the crude extracts of sparganlim from snake, Matrix tigrina laterolis and used as antigenic components. By SDS-PAGE, we detected a total 25 peptide bands (fractions) with 290 Kd to 23 Kd molecular weight, and 8 bands of these detected bands developed strongly by silver stain. In serological test, ELISA, we recognized the cross-reaction of antigenic components reacting with IgG antibodies in heterogenous sera, cysticercosis and hydatidosis patients sera. The crude antigenic components of sparganum showed the high sensitivity in sparganosis, hydatidosis patients sera, but showed lower sensitivity in cysticercosis patients sera than the sparasanosis, hydatidosis patients sera. Sixteen antigenic components of these 25 separated bands were recognized by antibodies In sparsanosis patients sera,8 antigenic components in normal human sera, 4 antigenic components in cysticercosis patients sera and 19 antigenic compoenents in hydatidosis patients sera. The crude antigenic compnenets with 290 Kd, 200 Kd, 125 Kd and 28 Kd molecular weight was only recognized in sparganosis patients sera, but 64 Kd antigenic component was nonspecific antigenic components which were also cross-reacted with sparganosis, hydatidosis, cysticercosis patients sera and normal human sera.

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Neutralizing Effects of Antiserum by Repeated Subcutaneous Administration of Recombinant Human Growth Hormone (rhGH)

  • Song, Yeon-Jung;Park, Shin-Hye;Park, Seung-Kook;Yeon, Je-Deuk
    • Biomolecules & Therapeutics
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    • v.10 no.3
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    • pp.175-179
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    • 2002
  • Human growth hormone (hGH) forms antibody by repeated administration. The present study investigated to confirm formation of antibody by repeated subcutaneous administration of hGH for two months in rats and dogs. In this result, hGH-injected sera were significantly higher than control sera by 1:1,000,000 of dilution factor. After antibody formed sera (anti-hGH sera) and control sera were added to 30 $\mu\textrm{g}$/ml hGR, the complex incubated for overnight at $30^{\circ}C$. Anti-hGH sera decreased hGH contents about 90% compared to control sera. Also, body weight gain conducted decreased about 67% compared to control sera in hypophysectomised rat. Inconclusion, repeated administration of hGH formed antibody because hGH was foreign protein to rats and dogs. And formed antibody of hGH was blocked and decreased many efficacy of hGH, the antibody was proved to be neutralizing antibody. Thus, because neutralizing antibodies were decreased pharmacological effects of hGH, administration more than two months were no significance.

Agglutinating Antibody to Propionibacterium acnes Serotype I and Serotype II in Normal Human Sera (정상인(正常人) 혈청(血淸)의 Propionibacterium acnes Serotype I 및 Serotype II에 대한 항체(抗體))

  • Choi, Chul-S.;Seo, Yang-Y.;Yang, Yong-T.
    • The Journal of the Korean Society for Microbiology
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    • v.14 no.1
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    • pp.63-69
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    • 1979
  • Antibodies to Propionibacterium acnes(Corynebacterium parvum) serotype I and serotype II in normal human sera were measured using a microtitre bacterial agglutination test. Of 168 sera tested, 53 sera(31.0%) exhibited higher agglutinin titres to serotype I than to serotype II and 34 sera(20.2%) gave higher titers to serotype II than to serotype I. Eighty-one sera(48.3%), however, showed similar antibody titres to both types. Antibodies to serotype I(x) and serotype II(y) showed high correlation(r=0.73, p<0.01) and regression equation was Y=1,078+0.73X. The mean antibody titre($log_2$) of 529 normal sera(male 447 and female 82) to serotype I was $5.49{\pm}1.36$, but there was no significant difference between male($5.45{\pm}1.36$) and female($5.74{\pm}1.36$). Bacterial agglutinin to Propionibacterium acnes in normal sera belonged to a 2-mercaptoethanol resistant IgG class.

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Use of the enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine (ELISA를 이용한 돼지 톡소플라스마병의 조기 진단에 관한 연구)

  • Suh, Myung-deuk;Jang, Dong-hwa;Joo, Hoo-don
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.567-575
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    • 1989
  • This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

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Bactericidal Activity of Normal and Cirrhotic Patients' Sera against Vibrio vulnificus (정상인 및 간경화환자 혈청의 Vibrio vulnificus에 대한 살균효과)

  • Rhee, Joon-Haeng;Chung, Sun-Sik
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.121-126
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    • 1986
  • Bactericidal activity of normal and cirrhotic patients' sera against Vibrio vulnificus was investigated and compared each other as a part of the pathogenesis study of V. vulnificus septicemia. Bactericidal activity of serum against V. vulnificus was complement-dependent and the alternative pathway played the major role. V. vulnificus strains CDC A1402 and ATCC 27562 appeared serum-sensitive, and CDC C7184, clinical isolate CNUH1 and ATCC 29307 were serum-resistant. When bactericidal activity of cirrhotic sera against serum-sensitive CDC A1402 and serum-resistant CDC C7184 was compared with that of normal sera. showed slightly poorer bactericidal activity than normal sera. Especially in the case of serum-resistant CDC C7184 strain, cirrhotic sera showed statistically significant decrease(p<0.05) in their bactericidal activity.

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