• Title/Summary/Keyword: Antibody specificity

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Preliminary Studies on the Measurement of the Free Thyroxine in the Serum by the Radioimmunoassay (방사면역측정법(放射免疫測定法)에 의한 혈중(血中) 유리(遊離) Thyroxine 농도측정(濃度測定)에 대(對)한 기초적(基礎的) 검토(檢討))

  • Lee, Joon-Il
    • Journal of radiological science and technology
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    • v.10 no.1
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    • pp.85-90
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    • 1987
  • Preliminary studies on the measurement of the free thyroxine in the serum with Amerlex $FT_{4}$ RIA kit were investigated using a tracer as $^{125}I-T_{4}$ derivative which is not almostly bound to thyroxine binding globulin, etc. The results are followed: 1. Linearity was tested on standards at various concentrations, and reproducibility and accuracy was excellent. 2. The antibody specificity is also excellent, and standard calibration curve of total $T_{4}$ was similar that of adding the TBG inhibitor. 3. Each value of $T_{4}$ in serum (the normal group, the hypothyroidism patients the pregnant women and the TBG dificiency patients) was not significant. As mentioned above, this method is more simple and rapid, compared to the other method.

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Enoylpyruvate Transferase Isozymes in Bacillus megaterium

  • Choi, Seung-Tae;Katsuji Tani;Ryoka Matsuno
    • Journal of Life Science
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    • v.2 no.4
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    • pp.232-239
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    • 1992
  • UDP_GlcNAc is metabolized to form vegetative cell wall, cortical peptidoglycans, and outermost layer consisting of galactosamine-6-phosphate ploysaccharide in life cycle of Bacillus megaterium. To obtain a better understanding of the UDP-GlcNAc regulation, we examined the activity of the common first enzyme for the synthesis of nucleotide precursors of peptidoglycans, enoylpyruvate transferase by newly developed method. Both the specific and the total activity decreased after the end of exponential growth followed by and increase from t5 but decreased again parallel to the appearance of the activity of UDP_GlcNAc-4-epimerase. Antibody specificity to anti-transferase IgG and the elution profile on DEAE-Sepharose revealed that B. megaterium has at least two enoylpyruvate transferase isozymes, and UDP_GlcNAc was metabolized to vegetative cell wall and cortical peptidoglycan by each isozme in exponential growth and in sporulation, respectively in life cycle.

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Development of an ELISA for the Organophosphorus Insecticide Chlorpyrifos

  • Cho, Young Ae;Lee, Hye-Sung;Park, Eun-Yeong;Lee, Yong-Tae;Hammock, Bruce D.;Ahn, Ki-Chang;Lee, Jae-Koo
    • Bulletin of the Korean Chemical Society
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    • v.23 no.3
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    • pp.481-487
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    • 2002
  • A selective enzyme-linked immunosorbent assay (ELISA) for the insecticide chlorpyrifos was developed. Four haptens for chlorpyrifos were synthesized and two of them were used as immunogens after coupling to keyhole limpet hemocyanin by two differe nt approaches. Rabbits were immunized with either of them and the sera were screened against 4 haptens coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigencoated ELISA was developed, which shows an I50 of 160 ppb with a detection limit of 10 ppb. An antibody-coated ELISA was also developed, which shows an $I_{50}$ of 20 ppb with a detection limit of 0.1 ppb. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except for insecticides chlorpyrifos-methyl and bromophos-ethyl, which makes these assays suitable for the selective detection of chlorpyrifos.

Functional Display of Maackia amurensis Hemagglutinin (MAH) on Bacteriophage (박테리오파아지 표면 발현 시스템을 이용한 Maackia amurensis Hemagglutinin (MAH)의 기능적 발현)

  • 임미정
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.176-179
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    • 2003
  • A library of unlimited number of novel lectins with diverse specificities has been previously generated by randomly mutating the carbohydrate-recognition domain of Maackia amurensis hemagglutinin (MAH). To establish the experimental environment capable of selecting high affinity mutant lectins in E. coli, phage display system was adapted. Carbohydrate binding capacity of two phagemid vectors, pComb3 and pComb8 displaying wild-type MAH lectin was assessed. Specific bindings of pComb3 and pComb8 phages expressing w.t. MAH to affinity-purified polyclonal anti-MAH antibody and to glycophorin was demonstrated. Both phages also showed strong hemagglutinating activity to intact but not sialidase-treated human erythrocytes, which is consistent to the specificity of native MAH. Taken together, two different phage display vectors successfully allowed the expression of active MAH as a fusion protein on the surface of bacteriophage, which will lead to preparation of unique plant lectins with high affinity toward a variety of carbohydrate chains.

Development of an ELISA for the Organophosphorus Insecticide Isofenphos

  • Park, Han-Jin;Park, Won-Chul;Jung, Tae-Owan;Rha, Choon-Sup;Lee, Yong-Tae
    • Bulletin of the Korean Chemical Society
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    • v.23 no.4
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    • pp.599-603
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    • 2002
  • A selective enzyme-linked immunosorbent assay (ELISA) for the insecticide isofenphos was developed. Three different analogues (haptens) of isofenphos were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group t o use as immunogens or coating antigens. Rabbits were immunized with one of the haptens coupled to BSA for production of polyclonal antibodies and the sera were screened against each of the other two haptens coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 96 ng/mL with the detection limit of 2 ng/mL. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides and the phenol metabolite of isofenphos, which makes the developed assay suitable for the selective detection of isofenphos. An antibody-coated ELISA was also developed, which showed an I50 of 580 ng/mL with a detection limit of 70 ng/mL.

Development of an Enzyme-Linked Immunosorbent Assay for the Organophorus Insecticide Bromophos

  • Park, Won-Cheol;Cho, Young-Ae;Kim, Yoo-Jung;Hammock, Bruce D.;Lee, Yong-Tae;Lee, Hye-Sung
    • Bulletin of the Korean Chemical Society
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    • v.23 no.10
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    • pp.1399-1426
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    • 2002
  • A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos. Three bromophos analogues (haptens) were synthesized and were coupled to carrier proteins to use as immunogens or coating antigens. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin (BSA) for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin (OVA). Using the serum with highest specificity and an enzyme tracer, an antibody-coated ELISA was developed, which showed an $IC_{50}$ of 40 ng/mL with a detection limit of 7 ng/mL. The antibodies in this assay showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides chlorpyrifos and fenitrothion.

Immunomodulatory activity of a polysaccharide isolated from cell culture of Acantopanax senticosus

  • Ahn, Hyo-Jung;Han, Sang-Bae;Yoon, Yeo-Dae;Lee, Haeng-Soon;Moon, Jae-Sun;Lee, Chang-Woo;Lee, Ki-Hoon;Park, Song-Kyu;Kim, Hyung-Chin;Kim, Hwan-Mook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.259.1-259.1
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    • 2002
  • Many polysaccharides isolated from plants are considered to be biological response modifiers and have been shown to enhance various immune responses in vivo and in vitro. Here we demonstrate that a polysaccharide isolated from cell culture of Acanthopanax senticosus (AS) has a unique mode of immunostimulation with regard to its cell-type specificity. AS was found to markedly increase polyclonal IgM antibody production and the proliferation of B cells. and to activate iNOS transcription and NO production in macrophages. (omitted)

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Selective B cell activation by polysaccharide isolated from the root of Acanthopanax koreanum

  • Han, Sang-Bae;Ahn, Hyo-Jung;Yoon, Yeo-Dae;Kim, Young-Ho;Lee, Jung-Joon;Moon, Jae-Sun;Lee, Chang-Woo;Lee, Ki-Hoon;Park, Song-Kyu;Kim, Hyung-Chin;Kim, Hwan-Mook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.257.1-257.1
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    • 2002
  • Many polysaccharides isolated from plants have been shown to enhance various immune responses in vivo and in vitro. Here we demonstrate that polysaccharide isolated from the root of Acanthopanax koreanum (AK) has a unique mode of immunostimulation with regard to its cell-type specificity. AK was found to markedly increase polyclonal 1gM antibody production and the proliferation of B cells. (omitted)

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An epidemiological study of human paragonimiasis by means of micro-ELISA (면역효소진단법을 이용한 폐흡충증 유행의 역학조사)

  • 조승열;이동근강신영김석일
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.246-256
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    • 1983
  • As epidemiological parameters of human paragonimiasis, the positive rates of intradermal test and the sputum/stool ekaminations have long been employed in population surveys. However, both the specificity of the intradermal test and the sensitivity of sputumjstool examination have been gradually declined as the endemicity was lowered; thus the gap between above two parameters widened. In such context, the development of a new epidemiologic parameter or tool which makes it possible to accurately discriminate the active paragonimiasis cases was necessary. In the present study, the detection rate of Paragonimus-speclac IgG antibody by micro-ELISA was evaluated as an indicator of epidemiologic status of human paragonimiasis in a population. A total of 4, 285 students and inhabitants living in Bukpyeong Myeon and Bukil Myeon, Haenam Gun, Jeonlanam Do was surveyed in October, 1983 by intradermal test first. Out of them, 244 cases (5.7%) were found positively reacted to VBS antigen of F. westermani. Out of 168 positive reactors, 7 cases (4.2%) were egg positive either by two times of sputum examination or by one stool examination. That indicated that only 0. 16% of total surveyed were confirmed as active paragonimiasis by egg detection. When sera collected from 239 positive reactors of Intradermal test were tested by micro-ELISA for their specific IgG antibody, 40 cases(16.7%) were found to be positive. All of 7 egg positive cases were again positive for specific IgG antibody. Among remaining 232 intradermal test positive cases, 33 cases were positive for IgG antibody. In contrast to those, none of 42 positive reactors to intradermal test for Cloncrchis and of 128 intradermal test negative cases showed positive for Paragcnimus-specIfic IgG antibody. The rate of specific IgG antibody as detected by micro-ELISA appeared to be increased with the wheal size of the intradermal test. When the wheal sixte was over 13mm in diameter, about 50% of them were positive for specific IgG antibody. Thirty-one specific antibody positive cases were clinically evaluated by laboratory examinations (repeated sputum examination, peripheral eosinophil count and chest roentgenogram) and by history taking. Out of them 24 cases were associated with one or more positive laboratory findings: thus considered as active paragonimiasis cases. Out of 7 lab. finding-free cases 3 revealed past history of typical paragonimiasis symptoms, suggesting that they were in chronic or in convalescent stages. The remaining 4 cases were considered as either mild or ectopic infection cases; the possibility of cross-reaction with other helminthiases could not be ruled out. From the above results, it was inferred that the detection of Paragonimus-specIfic IgG antibody by micro-ELISA was very much helpful in detecting the active cases as well as in proper evaluation of the endemicity of human paragonimiasis in a population. The convenience of mass haildling of sera in micro-ELISA was considered another superiority as an epidemiologic tool.

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Evaluation of the antibodies against Pseudomonas tolaasii and Pseudomonas agarici for Immunoassays and Its Application (항 Pseudomonas tolaasii 및 Pseudomonas agarici 항체의 면역측정법을 위한 평가와 그 이용)

  • Jeon, Nak-Beom;Lee, Hyang-Burm;Shon, Dong-Hwa;Yu, Seung Hun
    • Research in Plant Disease
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    • v.9 no.2
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    • pp.94-98
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    • 2003
  • To produce antibodies against Pseudomonas tolaasii and P. agarici, lyophilized P. tolaasii and P. agarici ($5{\times}10^7$ cfu/ml) and Freund, s adjuvant were immunized into rabbits 4 times. The specificity and sensitivity of the antibodies were evaluated by immunodiffusion test and indirect enzyme-linked immunosorbent assay (id ELISA). The ${\alpha}$-P. tolaasii antibody was very specific only against P. tolaasii, while ${\alpha}$-P. agarici antibody was not specific and showed a high cross reactivity toward P. tolaasii with detection limit concentration of $2{\times}10^3$ cfu/ml. However, the cross reactivities of ${\alpha}$-P. agarici antibody toward the related species including P. reactans were very low. Our results showed that ${\alpha}$-P. tolaasii and ${\alpha}$-P. agarici antibodies against P. tolaasii and P. agarici, respectively, might be useful for rapid and simple detection of the causal agents of bacterial brown and yellow blotches in cultivated oyster mushrooms.