• Title/Summary/Keyword: Coating tracer

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The optimization of ELISA for methamphetamine determination : the effect of immunogen, tracer and antibody purification method on the sensitivity

  • Choi, Jeongeun;Choi, Myung-Ja;Kim, Choonmi;Cho, Young-Shik;Chin, Jaeho;Jo, Young-Ah
    • Archives of Pharmacal Research
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    • v.20 no.1
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    • pp.46-52
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    • 1997
  • To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein G-purified antibodies. The cross-reactivity of the purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations.

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Microcapsules Containing Self-Healing Agent with Red Dye (빨간 색소를 함유한 자가치료제 마이크로캡슐)

  • Guang, Yang;Lee, Jong Keun
    • Polymer(Korea)
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    • v.37 no.3
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    • pp.356-361
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    • 2013
  • Microcapsules of two different self-healing agents, 5-ethylidene-2-norbornene (ENB) and ENB with a crosslinker, surrounded by a melamine-urea-formaldehyde shell were manufactured. In this work, a red dye was incorporated into the self-healing agents as a tracer for better visual observations. It revealed that the incorporation of a red dye into self-healing agents did not disturb the formation of microcapsules from the examination of thermal resistance, particle size/size distribution and morphology of the resulting microcapsules. Releasing of self-healing liquid into the induced crack from ruptured microcapsules and filling between crack planes were observed using an optical microscope. Also observed was the reaction of filled healing agent with embedded Grubbs' catalyst in an epoxy coating layer.

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.

Development of the Micro Metal Balloon Using Sirasu-balloons as a Core Material

  • Uezono, Tsuyoshi;Sodeyama, Ken-ichi;Onomae, Hiroshi;Sakka, Yoshio
    • Proceedings of the Korean Powder Metallurgy Institute Conference
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    • 2006.09a
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    • pp.604-605
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    • 2006
  • Recently the Marangoni convention is supposed to be an important phenomenon that significantly affects the solidification. For understanding the Marangoni convection mechanism, visualizing the convention phenomenon of molten tin with ultrasonic has been conducted. This paper reports developing a tracer material of micro metal balloon that is used in the molten system. We have succeeded in coating the surface of Shirasu-ballons with nickel by plating process. The obtained metal balloon is spherical and some characterizations were conducted.

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Enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus (축우 부루셀라병의 ELISA 진단법에 관한 연구)

  • Lim, Yoon-kyu;Lee, Doo-sick;Park, Jun-hong;Yang, Ki-chun;Kim, Seung-ho;Kim, Kong-sick;Hyun, Kwan-jong;Kim, Woo-tack;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.131-135
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    • 1993
  • Enzyme-linked Immuno sorbent Assay (ELISA) for the serological diagnosis of Brucella abortus was developed and compared with plate agglutination test. Cell wall antigen was extracted from Brucella abortus 1119-3 by sonication and with a sodium deoxychlate solution. Optimum protein concentration of coating antigen were $0.4{\mu}g/100{\mu}{\ell}$ protein on each microtiter plate well. Horse radish peroxidase (HRP) labeled protein-G was used as a tracer of reacted antibodies. ELISA confirmed the agreeable results of 40 cases out of 43 cases by plate aggulutination test. ELISA diagnosed positive cases(10 out of 12) and negative cases (1 out of 12) with dubious sera by plate agglutination test. From this results ELISA could be used for the early diagnostic tools of Brucellosis in cattle.

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Optimization of Immunosasay Procedures for the measurement of Progesterone (Progesterone 측정을 위한 면역분석법의 최적조건에 관한 연구)

  • 강원준;고대환;이경광;김종배;정길생
    • Korean Journal of Animal Reproduction
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    • v.9 no.2
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    • pp.105-112
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    • 1985
  • This research was carried out to investigate the optimal conditions ssociated with the RIA procedures such as a bridging phenomena, prozone effects and a new separation methods etc. The results obtained were summarized as follows: 1. The lgG fractions of donkey anti-rabbit IgG sera were purified by Protein-A-Sepharose affinity column, which indicates that Protein-A an affinity for IgG class of donkey antiserum. 2. In coating the IgG fraction on polystryene tubes, incubation conditions made no differences between 2 hr at room temperature and overnight at 4$^{\circ}C$. 3. There were no significant differences between 1st antibody-coated tube and 2nd antibody-coated tube as a separation method when compared in terms of reproducbility. A better reproducibility may be expected if the titers of 1st antibody for the progesterone to be assayed and of corresponding 2nd antibody are reasonably high. 4. The titers of anti-progesterone antibody for 3H-progesterone and progesterone-11HS-125I were 1:300 and 1:700 in liquid-phase, and 1:100 and 1:300 in solid-phse for the separation methods. 5. A bridging phenomena in which a standard curve is long and shallow were observed when progesterone-11HS-125I was used for the tracer, but not in 3H-progesterone. 6. A prozone effect in a solid-phase system, especially 1st antibody-coated tube method was observed which the degree of inhibition was significantly different although zero bindings look the same. In this case, the titration curve should be made both in the absence and in the presence of a, pp.opriate amount of competiter, standard, respectively.

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