• Title/Summary/Keyword: Antisera

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The serodiagnosis of a lymphocystis disease virus infection using an antibody raised against a recombinant major capsid protein

  • Seo, Ja-Young;Kang, Bong-Jo;Oh, Hyoung-Jong;Lee, Jae-Il;Kim, Tae-Jung
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.175-180
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    • 2008
  • Lymphocystis is a viral disease of fish primarily in marine and brackishwaters. Here we report the cloning, expression, and the serological applications of the lymphocystis disease virus (LCDV) major capsid protein (MCP). The MCP gene was amplified by PCR from the genomic DNA of LCDV isolated from Schlegel's black rockfish, Sebastes schlegeli, and expressed in E. coli. Mouse antisera raised against the purified recombinant MCP (rMCP) reacted with the viral MCP in an immunofluorescence assay, indicating that this rMCP would be useful for serological studies of field samples.

Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Insecticide Cyanophos

  • Park, Jae-Hyun;Park, Won-Chul;Kim, Yoo-Jung;Lee, Yong-Tae
    • Bulletin of the Korean Chemical Society
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    • v.23 no.4
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    • pp.605-609
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    • 2002
  • A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of organophosphorus insecticide cyanophos. An analogue (hapten) of cyanophos was synthesized and was coupled to BSA to produce polyclonal antibodi es from rabbits. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 310 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivities with other organophosphorus pesticides except for parathion-methyl, which makes the assay suitable for the selective detection of cyanophos.

Biological, Physico-chemical and Serological Characteristics of TMV Strains Isolated from Tobacco, Tomato and Pepper Plants (담배, 토마토 및 고추에서 분리된 TMV 계통의 생물학적, 물리화학적 및 혈청학적 특성)

  • 박은경;이청호;이영기;김영호
    • Journal of the Korean Society of Tobacco Science
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    • v.19 no.1
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    • pp.5-10
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    • 1997
  • Three strains of W isolated from tobacco, tomato and Pepper plants in Korea were characterized based on biological response, serological relationship, and peptide mapping of the capsid Proteins. The strains designated as TMV-common, TMV-Pepper, and TMV-tomato could be distinguishable by different visual symptoms on 3 varieties of tobacco, one variety of tomato and Pepper for each among 27 plant specieces. Serological relationships were examined by agar gel double diffusion test. Only traceable or weak reaction was observed in the incompatible antigen-antibody combinations. The Pepper strain, however, showed trace in reaction with other two antisera. Peptide maps of the capsid proteins digested by V8 protease or by trypsin were also distinguishable, suggesting differences in composition and/or sequence of the amino acids among the strains.

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Physicochemical Study on the Synthesis of Progesterone Immunogen (프로제스테론의 면역원 합성에 관한 물리화학적 연구)

  • Park, Jun-hong;Kwun, Jong-kuk
    • Korean Journal of Veterinary Research
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    • v.26 no.2
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    • pp.225-228
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    • 1986
  • Progesterone immunogen has been synthesized and its melting point, Rf-value, UV and IR spectrum have been measured to develope the essential step in antisera production against low molecular weight substance. Mixed anhydride reaction was used to conjugate $11{\alpha}$-hydroxy-progesterone with succinic anhydride. Melting point of one intermediate compound was $156^{\circ}C$, and Rf-value was 0.41 in benzene : acetone : methanol (5 : 5 : 2). Maximum absorbance was measured at 242nm and ${\varepsilon}$ was $1.641{\times}10^4cm^2/mole$. Loss of hydroxy group was observed at 3450nm, and carbonyl group was appeared at 1160nm, 1250nm and 2960nm. These results indicated that the intermediate compound was progesterone hemisuccinate. Maximum absorbance of progesterone bovine-serum albumin(BSA) conjugate was observed at 250nm. Molar ratio of progesterone to BSA was average 15.4 on UV spectrum.

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Immunohistochemical study on the endocrine cells of the pig stomach (돼지 위점막의 내분비세포에 관한 면역조직화학적 연구)

  • Lee, Jae-hyun;Kim, Jeong-mi;Lee, Hyung-sik
    • Korean Journal of Veterinary Research
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    • v.37 no.1
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    • pp.1-8
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    • 1997
  • The relative frequency and distribution of occurrence of immunoreactive cells in the proventriculus, diverticulum, cardia, fundus and pylorus of the stomach of pigs were investigated by PAP method using specific antisera against BCG, Gas/CCK, 5-HT, somatostatin, glucagon, BPP, motilin and insulin. In the diverticulum and cardia, BCG-, 5-HT-, somatostatin- and glucagon-immunoreactive cells were detected. In the fundus, BCG-, 5-HT- and somatostatin-immunoreactive cells were also found. In the pylorus, BCG-, Gas/CCK-, 5-HT-, somatostatin- and glucagon-immunoreactive cells were observed. However, no BPP-, motilin- and insulin-immunoreactive cells were found in the stomach epithelium of the pigs. These results showed that the occurrence of the endocrine cells confirmed in the diverticulum as the cardia and suggest that the function of diverticulum may be similar to that of cardia in the pigs.

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The Characteristics of Fish Metallothionein and Its Application to the Biomonitoring for the Evaluation of Water Pollution (어류 Metallothionein의 툭성 및 수질오염 평가를 위한 생물모니터링에의 응용)

  • 황갑수
    • Environmental Analysis Health and Toxicology
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    • v.12 no.3_4
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    • pp.15-22
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    • 1997
  • This experiment was performed to examine the immuno-reactive characteristics of fish metal-binding protein, metallothionein (MT), and gain the practical understandings for the proposed use of fish MT as a biomarker. Liver MT induced by Cd in the silver carp was seperated and purified by gel filtration chromatography and ion exchange chromatography. The immuno-reactivity of fish MT was examined with 3 rabbit antisera. Fish MT showed little reactivity with rabbit anti-rat MT antiserum and a weak reactivity with anti-MT peptide antiserum while showed a strong reactivity with rabbit anti-fish MT antiserum. The time-course change of liver MT in the silver carp, after waterborne exposure to 1 ppm of Cd, was checked by Cd-hem method and established competitive ELISA. In both cases, the induction of liver MT showed a good increasing relationship with the exposure days. The results indicate that the fish MT can be developed as a useful biomonitoring means in the toxicological study and for the evaluation of water pollution.

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Isolation and Partial Chemical Characterization of the Yolk Proteins from Drosophila sp. (robusta species group) (Drosophila sp.(robusta species group)의 난황 단백질의 분리 및 부분적 화학적 특성)

  • Kim, Se-Jae;Gi
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.17-22
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    • 1992
  • The three yolk polypeptides have been isolated and partially characterized. Their molecular weights of YPI, YP2, and YP3 were 48, 000, 47, 000, and 46, 000, respectivelv, as judged by SDS-polyacrvlamide gel electrophoresis. They have different digestion products upon in situ peptide mapping by limited proteolysis. Two-dimensional gel electrophoresis showed that their isoelectric points were heterogeneous from 5.92 to 6.54. And thew showed three different antigen-antibody reactions when each polvpeptides is reacted with antisera made to a mixture of all of three. These data reported here indicate that the yolk proteins are consisted of distinctive polypeptides in Drosophlla sp. (robusta species group).

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Induction of Cd-binding High Molecular Weight Protein [Cd-BP(II)] in Rat Tissues (흰쥐 조직에서의 카드뮴 결합 고분자량 단백질 [Cd-BP(II)]의 유도)

  • 천기정;김봉희
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.591-597
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    • 1999
  • The study was carried out on the biochemical characters of Cd-BP(II) after isolation and purification of the protein from the liver of rat ip injection with Cd. A continued study has been doing whether Cd-BP(II) could be induced by Cd or by the other metals such as Zn and Cu. Antisera were made against the antigen of Cd-BP(II) from New Zealand white rabbits. We carried out g-globulin purification, then Ouchterlony test and gel immunodiffusion test. Cd-BP(II) was also found in normal tissues of rat. It was induced up to a considerable level by Cd, whose induced level was higher than that Cu or Zn treatment. The level of induction by Cu or Zn pretreatment plus Cd treatment was lower than that by single treatment of Cu or Zn. Such a result was presumably related to the Cd toxicity.

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Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocyte for Producing Fat-reduced High Quality Pork (저지방 고품질 돈육 생산을 위한 돼지 복강 및 피하지방 항체 개발)

  • Choi, Chang-Weon;Baek, Kyung-Hoon;Choi, Chang-Bon;Oh, Young-Kyoon;Hong, Seong-Koo
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.87-94
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    • 2010
  • The aim of the present study was to develop polyclonal antibodies to regional inedible adipocytes of pigs and investigate the effect of these antibodies on adipocytes in vitro. As antigens, abdominal and subcutaneous adipocyte PMPs from pigs were injected into sheep 3 times per 3 wk intervals for passive immunization, and non-immunized serum, antisera against abodominal (AAb) or subcutaneous adipocyte PMPs (SAb) were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with the heart, kidney, liver, lung, muscle, and spleen of pig were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes from pigs were performed to analyze LDH concentration. At a 1:1,000 dilution, little antibody reactivity was observed for non-immunized serum whereas both AAb and SAb had relatively strong reactivity up to a dilution of 1:16,000. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivity of AAb and SAb was detected with the PMPs of the organs. Both antisera most strongly reacted with each adipocyte PMPs and showed statistically (p<0.05) higher cross-reactivities compared with the non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and are safe against cross-reactivities with the organs of pigs. Further studies on the in vivo nutritional safety and fat reduction of these antibodies in pigs will be required fat-reduced high quality pork production.

Expression, Purification and Antiserum Production of the Avian Influenza H9N2 Virus HA and NA Proteins (Avian Influenza H9N2 Virus의 HA와 NA 단백질 발현, 정제 및 항혈청 생산)

  • Lee, Hyun-Ji;Song, Byung-Hak;Kim, Jeong-Min;Yun, Sang-Im;Kim, Jin-Kyoung;Kang, Young-Sik;Koo, Yong-Bum;Jeon, Ik-Soo;Byun, Sung-June;Lee, Youn-Jeong;Kwon, Jun-Hun;Park, Jong-Hyeon;Joo, Yi-Seok;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.178-185
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    • 2008
  • Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.