• Title/Summary/Keyword: Thiolation

Search Result 13, Processing Time 0.026 seconds

Detection of IgG Using Thiolated Protein G Modified SPR Sensor Chip (Thiolated protein G로 개질된 SPR 센서 칩을 이용한 IgG 검출)

  • Sin, Eun-Jung;Lee, Yeon-Kyung;Sohn, Young-Soo
    • Journal of Sensor Science and Technology
    • /
    • v.20 no.6
    • /
    • pp.434-438
    • /
    • 2011
  • A portable surface plasmon resonance(SPR) based immunosensor using thiolated protein G and protein G was developed for the detection of immunoglobulin G(IgG). The protein G has specific affinity with Fc fragment of IgG and was thiolated by 2-Iminothiolane for introduction of thiol groups. Anti-IgG, bovine serum albumin(BSA), and IgG have been sequently injected after surface modification of gold sensor chip with protein G and thiolated protein G. The output signal was increased with the injection of each protein and the actual signal was measured by subtracting signal of reference channel from signal of sample injected channel. The experimental results showed the higher detection capability of IgG using thiolated protein G compared with protein G. From these results, we can conclude that the current surface modification technique and the portable SPR sensor system can be applied to various immunosensors for diagnosis.

Induction of Oxidative Stress and Cytoskeleton Damage by Cadmium in WB-F344 Rat Liver Epithelial Cells (랫드간장상피세포에서 카드뮴에 의한 산화적 스트레스 및 Cytoskeleton 손상 유발에 관한 연구)

  • 정상희;조명행;조준형
    • Toxicological Research
    • /
    • v.14 no.4
    • /
    • pp.577-585
    • /
    • 1998
  • Cadmium is an important industrial and environmental pollutant and has adverse effects on cell growth and metabolism, although the mechanisms of its cellular toxicity are still unclear. This study was performed to elucidate the cytotoxic mechanism of cadmium in the viewpoint of oxidative stress and cytoskeleton alterations in WB-F344 rat liver epithelial cells. 200 $\mu\textrm{M}$ $CdCl_2$ caused a severe disassembling of microtubule and micro filament and an apparent cell retraction under an observation with fluorescence micoscope. (equation omitted)-tubulin and F-actin protein were highly thiolated at 20 min and then disappeared from 1 hour after the treatment of 200 $\mu$M CdCl$_2$in the immunoblot analysis. Intracellular GSH was decreased from 1hr to 24 hrs by 66.6 or 200 $\mu\textrm{M}$ of $CdCl_2$. Intracellular protein thiol was also decreased by 22.2, 66.6 and 200 $\mu\textrm{M}$ of $CdCl_2$ at 1 hour after its treatment. The product of lipid peroxidation (malondialdehyde) was increased from 4 hrs by 66.6 and 200$\mu\textrm{M}$ of $CdCl_2$. These data indicate that cadmium induces oxidative stress involving disassembling of microtubule and micro filament, thiolation of (equation omitted)-tubulin and actin protein, depletion of GSH and protein thiol, and increase of lipid peroxidation.

  • PDF

Detection of Carp Vitellogenin with Piezoelectric Immunosensor (압전류적 면역센서에 의한 잉어 Vitellogenin 검출)

  • Kim, Nam-Soo;Park, In-Seon;Kim, Woo-Yeon
    • Applied Biological Chemistry
    • /
    • v.49 no.3
    • /
    • pp.254-258
    • /
    • 2006
  • Vitellogenin has been known as a potent biomarker protein for the estrogenic activity in fish exposed to endocrine disruptors. In this study, a piezoelectric immunosensor making use of an anticarp vitellogenin antibody and an AT-cut quartz crystal microbalance as the biological component and transducer was prepared, followed by its application to the analysis of carp vitellogenin as follows. Antibody immobilization was conducted by chemisorption of a thiolated antibody with a heterobifunctional thiolation cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. The reaction buffer for the immunosensor system was optimized as 0.1 M sodium phosphate (pH 7.4). Concentration-dependent sensor responses were obtained in the vitellogenin concentrations ranging from 0.4864 to 486.4000 nM, with a linear correlation between vitellogenin concentration and frequency shift in double-logarithmic scale. The limit of detection of the immunosensor for carp vitellogenin was presumed as 0.4864 nM.