• Title/Summary/Keyword: ELISA antibody

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Conformation of Group "a" Epitope in Hepatitis B Surface Antigen

  • Chun, Mun-Ho;Park, Won-Bong;Bok, Jin-Woo;Kim, Ha-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • v.15 no.4
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    • pp.347-355
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    • 1992
  • To elucidate structure of group "a" epitope, mouse antibodies that express idiotype monoclonal antibody and anti-idiotype monoclonal antibody against the group specific "a" determinant were purified by hydroxyapatite column. To obtain hepatitis B surface antigens (HBsAg). HBsAg positive blood was sequencially purified by ammonium sulfate precipitation, hydroxyapatite, sepharose 4B column chromatography and ultracentrifugation. The major protein (p25) and glycoprotein (gp30) of HBsAg were isolated by concanavalin-A-sepharose 4B. The ability of p25-gp30 among the HBsAg to inhibit the idiotype-anti-idiotype reaction was dependent on conformation, since reduced and alkylated p25-gp30 virtualy lost their inhibitory capacity when compared to native HBsAg. The data suggest that hepatitis B antigen is a conformational antigen critically dependent upon the disulfide bonds of p25-gp30.

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Comparison of immunogenecities of three beta-nodavirus proteins, capsid protein, non-structural protein B1 and B2 in olive flounder

  • Cha, Seung-Ju;Do, Jeong-Wan;Ko, Myoung-Seok;Kim, Jin-Woo;Park, Jeong-Woo
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.219-228
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    • 2009
  • The genomic and subgenomic RNAs of fish nodavirus encode the four proteins, protein A, capsid protein, non-structural protein B1 and B2. In this study, we describe the immune response of olive flounder Paralichthys olivaceus immunized with live fish nodavirus or recombinant capsid protein, non-structural protein B1 and B2 expressed in E. coli. Nodavirus-infected flounder produced antibodies to capsid protein, B1 and B2 and nodavirus-neutralizing activities were detected in the serum of the nodavirus-infected flounder. The flounder were immunized against the three recombinant proteins of fish nodavirus and the sera from these immunized fishes were assayed for nodavirus-specific antibody by ELISA and a neutralization test. In the immunized flounder, all three recombinant proteins induced the production of similar levels of antibody, but only the antibody to capsid protein significantly neutralized nodavirus. These results indicate that all three nodaviral proteins are immunogenic in flounder, but only the capsid protein can induce neutralizing antibody against nodavirus.

Effects of Socheongryong-Tang on Immunoglobulin Production in Asthmatic Mice (소청용탕(小靑龍湯)이 천식(喘息)이 유발된 생쥐의 면역 글로불린 분비에 미치는 영향)

  • Jung, Sun;Cho, Su-Jin;Moon, Kyoung-Il;Kim, Hyung-Woo;Kim, Bu-Yeo;Cho, Su-In
    • The Korea Journal of Herbology
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    • v.23 no.1
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    • pp.23-28
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    • 2008
  • Objectives : Socheongryong-Tang(小靑龍湯, SCRT), a herbal remedy, has been widely used to treat respiratory disease such as cough and asthma in Oriental countries. Recent years SCRT was known as anti-allergic agent. However, its therapeutic mechanisms including immunoglobuline such as IgE, IgG1, IgG2a productions are unclear. Methods : We investigated the effects of SCRT on levels of antigen specific total antibody, IgE, IgG1, IgG2a using ELISA method in serum from allergen-induced asthma mice. Results : SCRT decreased level of antigen specific IgE significantly. And SCRT treated mice showed downward tendency of IgG1, a Th1 relative antibody, level. But, SCRT did not affect levels of antigen specific total antibody and IgG2a, a Th1 relative antibody. Conclusions : we demonstrated the strong possibility of SCRT as a complementary or alternative drug to western drug also demonstrated that regulation of Th1/Th2 imbalance may be one of mechanism contributed to treatment for respiratory disease by SCRT.

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Immunological Properties of Recombinant Hepatitis B Surface Antigen Expressed in Mammalian Cell(C127)

  • Lee, Young-Soo;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.5
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    • pp.543-548
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    • 1998
  • We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross-immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. from the measurement of $ED_{50}$ after formalin- or heat-inactivation, both r-HBsAg and p-HBsAg and p-HBsAg showed $ED_{50}$ of 0.2-0.3 in formalin-inactivaton, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat-inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with $AIPO_{4}$ adjuvant. the mutagenicity of r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity.

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Production of monoclonal antibody to 45 kDa somatic protein of Trichuris suis (돼지편층의 45kDa 항원단백질에 대한 단클론항체 생산)

  • Lee, Jong-Kyung;Kim, Jung-Tae;Seo, Hun-Su;Park, Jong-Yeol;Yun, Hee-Jeong
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.625-635
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    • 2004
  • Trichnuris suis does not excrete eggs during larval stage as well as in particular adult stage, It is impossible to diagnose by use of fecal examination method in those periods. Therefore, serological diagnostic method can be very useful for those stages. In order to produce monoclonal antibody, specific somatic and secretory-excretory (SE) antigens of T. suis were identified and analyzed by SDS-PAGE and Western blot. Monoclonal antibody-producing hybridoma cells were cloned, which were made of popliteal lymph node of BALB/c mice immunized with a 45 kDa somatic antigen of T. suis. Five clones (1B9, 2C4, n2C5, 2D7 and 2D8) showing strong responses to T. suis antigens were selected and the isotype identified. All monoclonal antibodies were IgG1 isotype and the light chains were k chain. Established monoclonal antibodies reacted specifically to somatic and SE antigens of T. suis and did not cross-reacted to antigens of ascaris suum, trichuris vulpis, or Trichinella spiralis. The sensitivity of somatic and SE antigens against these monoclonal antibodies were significant (p<0.01) associated with those of positive and negative sera.

Seropositivity Rates of Strongyloides stercoralis Antibody in the Southeastern Region of Republic of Korea: A Single-Center Retrospective Study

  • Kim, Taehwa;Lim, Seungjin
    • Parasites, Hosts and Diseases
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    • v.60 no.3
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    • pp.181-185
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    • 2022
  • Strongyloides stercoralis infection is not endemic in the Republic of Korea (Korea) with a positivity rate of <1% in stool examination. However, there is a risk of hyperinfection in immunosuppressed individuals. It is necessary to determine the seropositivity of S. stercoralis antibodies in Korea. This study investigated the seropositivity of S. stercoralis antibodies in the southeastern area of Korea. From January 2019 to June 2021, serum samples were collected from participants who visited the study center in the southeastern region of Korea for routine health check-ups. We determined serum levels of specific anti-Strongyloides IgG antibodies in 834 samples by enzyme-linked immunosorbent assay. We observed that 92 samples (11.0%) tested showed a positive response. The age of the participants was 51±10.7 years, and 43.4% of them were men. The antibody positivity rate based on the location of the participants' residence were 12.3% (Gyoungsangnam-do), 10.2% (Busan), and 10.1% (Ulsan), respectively. Total eosinophil count was associated with positive test results (154.8±152.0 per mm3 versus 202.1±178.9 per mm3, P=0.006). Logistic regression analysis revealed that blood eosinophil count, age above 50 years, and residence in Sacheon were factors associated with the positive status of S. stercoralis antibody. Our finding suggests that it is necessary to test for S. stercoralis in actual clinical settings in Korea.

Measurement of 150 kDa protein of Taenia solium metacestodes by antibody-sandwich ELISA In cerebrospinal fluld of neurocystlcercosls patients (뇌 유구낭미충증 각자 혈청 및 뇌척수액에서의 유구낭미충 낭액항원의 측정)

  • Cho, Seung-Yull;Kong, Yoon;Kim, Suk-Il;Kang, Shin-Yong
    • Parasites, Hosts and Diseases
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    • v.30 no.4
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    • pp.299-307
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    • 1992
  • An antigenic protein in cystic quid of Taenia solium metacestodes (CF) of 150 kDa was measured by antibody-sandwich ELISA in serum and cerebrospinal quid (CSF) of neurocysticercosis patients. Capture antibodies were rabbit antisera against CF (RACF) and a monoclonal antibody (MAb) against 150 kDa protein in CF. Lower limit of antibody-sandwich ELISA was 8 ng/ml of the protein. Except CF, no tested helminthes extracts reacted. Levels of the protein in 351 sera from 255 patients (55 surgery confirmed and 202 antibody and CT/MRl confirmed) were below sensitivity of the assay. Of 276 CSF from 212 patients, 31 samples (11.2%) showed positive findings. This assay, therefore, was not sensitive enough to be a diagnostic. Instead, the 150 kDa protein appeared in CSF in such situations as in 2 days after prasiquantel treatment, or as in a patient infected with a racemose cysticercus with degenerated cyst wall. Of cases whose follow-up CSF were assayed, 2 cases showed that the protein appeared intermittently, These results suggest strongly that appearance of free 150 kDa protein is associated with cyst wall rupture. In CSF which contained the 150 kDa protein over 61 ng/ml, the protein was recognized in 505-PAGE before and after immunoprecipitation.

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Development of an Indirect Enzyme-Linked Immunosorbent assay for Rapid Detection of Adulteration of Food Allergen Mackerel in Processed Marine Foods (수산가공식품 중 알레르겐 고등어 혼입여부 신속 검출을 위한 간접효소면역분석법의 개발)

  • Lee, Jeong-Eun;Kim, Ah-Yoon;Kim, Sol-A;Kim, Hyo-In;Park, Ji-Hye;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.185-192
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    • 2018
  • The purpose of this study was to develop an indirect enzyme-linked immunosorbent assay (indirect ELISA) based on a monoclonal antibody (MAb) that is specific to mackerel thermal stable-soluble protein (TSSP), that can be used for the rapid detection of mackerel in processed marine foods. Among the four MAbs (3A5-1, 2, 9, and 12) developed in previous studies, the 3A5-2 MAb that showed high specificity and sensitivity were selected and used to develop the indirect ELISA method. The detection range of the indirect ELISA was 0.02%-0.001% and the detection limit of 0.001% was shown. No cross-reaction to other marine products and food ingredients was observed by the indirect ELISA. Processed marine foods containing mackerel with ${\geq}0.3$ O.D. value at 405 nm were estimated as positive samples by the indirect ELISA. Therefore, the indirect ELISA can be used as a rapid and sensitive method to identify mackerel authenticity and adulteration in processed marine foods.

Production and Characterization of a Monoclonal Antibody against Human ${\beta}_2$-adrenergic receptor

  • Kang, Suk-Jo;Shin, Chan-Young;Song, Mi-Ryoung;Lee, Chung-Jae;Cheong, Jae-Hoon;Lee, Sang-Bong;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.5 no.4
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    • pp.344-350
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    • 1997
  • The purpose of the present study was to produce and characterize a monoclonal antibody against human ${\beta}_2$-adrenergic receptor. Male BALB/c mice were immunized with glutathione S-transferase (GST) fusion protein of the C-terminal portion of the human ${\beta}_2$-adrenergic receptor which was expressed in E.Coli. The immunized splenocytes were fused with myeloma SP2/0-Agl4 cells. The resulting hybridomas were screened for the production of a monoclonal antibody which can recognize human ${\beta}_2$-adrenergic receptor, and then subcloned by limiting dilution. The resulting monoclonal antibody was named as mAb$\beta$CO2. The mono-clonal antibody $\beta$CO2 was determined as IgM subtype and then purified by anti-mouse IgM-agarose affinity chromatography. The results of ELISA, Western blot, and immunocytochemistry showed that mAb$\beta$CO2 recognized human ${\beta}_2$-adrenergic receptor in the ${\beta}_2$-adrenergic receptor-GST fusion protein and human spider-moid carcinoma cell line A431 with highly specific immunoreactivity, The monoclonal antibody $\beta$CO2 may provide useful tools for the study of the $\beta$-adrenergic receptor of human and other species including rats.

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Production and Characterization of DDT Antibodies and Its Application to Enzyme Immunoassay: Relation of Response and Affinity to Coating Ligand

  • Hong, Ji-Youn;Kim, Jong-Hyun;Park, Song-Ja;Lho, Dong-Seok;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • v.24 no.11
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    • pp.1605-1608
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    • 2003
  • To development an immunodetection method for DDT, 1,1,1-trichloro-2,2-bis(4-chorophenyl)ethane (p,p'-DDT) and its metabolites (p,p'-DDA, p,p'-DDE, p,p'-DDD), five derivatives of DDT haptens have been synthesised and characterized as coating ligands for antibody evaluation. The appropriate lengths of linkers were introduced to investigate a matching pair of coating ligand and antibody. Among these hapten derivatives, 2,2-bis(4-chlorophenyl)acetic acid (DDA), 5,5-bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for its use as an immunogen. The bovine serum albumin (BSA) conjugates of these derivatives were prepared as a coating ligand for monoclonal antibody screening. Fifteen monoclonal antibody clones were screened using these probes. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP), in addition to the above hapten derivatives, were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for their use as coating ligands to measure the titration level of the antibody and the displacement of free analytes. The indirect competitive ELISA results indicate that the titration level and free analyte displacement were greatly influenced by the DDT derivatives and carrier proteins used. Three matching pairs of monoclonal antibodies and coating ligands were selected for the DDT immunoassay: antibody clone 1A3 and coating ligand DDA-OVA, 1A1 and DDHHAP-BSA, and 1A4 and DDHP-OVA.