• Title/Summary/Keyword: horseradish peroxidase

Search Result 140, Processing Time 0.028 seconds

Use of Plant Materials for Decontamination of Waste Water Polluted with 2,4-Dichlorophenol (2,4-Dichlorophenol로 오염된 폐수의 정화를 위한 식물체의 이용)

  • Lee, Jung-Eun;Park, Jong-Woo;Kim, Jang-Eok
    • Korean Journal of Environmental Agriculture
    • /
    • v.18 no.3
    • /
    • pp.292-297
    • /
    • 1999
  • This study was performed to estimate the possibility of use of plant materials as catalytic agents fur the decontamination of waste waters contaminated with organic pollutants by using 2,4-dichlorophenol(2,4-DCP) as a model pollutant. Plant materials containing high peroxidase activity were selected as catalysts for the removal of 2,4-DCP. Peroxidase activity, which plant materials were containing, was measured, and the greatest peroxidase activity was observed in shepherd's purse, followed by turnip, sweet potato, Chinese cabbage and white radish. The peroxidase activity in shepherd's purse was four times higher than that of horseradish purchased in U.S.A. Using shepherd' s purse and turnip, it was investigated the effect of various factors on the decontamination of 2,4-DCP through oxidative coupling. The removal of 2,4-DCP was extremely fast, and a maximal removal could be achieved within 3 min for shepherd' s purse and 15min for turnip. The pH range was from 3.0 to 8.0 and the amount of $H_2O_2$ added was 9mM when maximal removal was achieved(over 90%). No increasing removal of 2,4-DCP was observed due to increasing the amount of $H_2O_2$ added (over 9mM). The initial concentration affected the transformation of 2,4-DCP incubated with plant materials. When turnip was used as catalytic agent, it was observed decreasing transformation of 2,4-DCP due to increasing initial concentration.

  • PDF

In Situ Identification of Cyanobacteria

  • Ahn Tae-Seok;Hong Sun-Hee;Chung Hyun-Mi;Belkova Natalia L.
    • Proceedings of the Microbiological Society of Korea Conference
    • /
    • 2001.11a
    • /
    • pp.121-127
    • /
    • 2001
  • Seven cyanobacteria strains (Anabaena macrospora NIERl0016, Oscillatoria sp. NIER10042, Microcystis aeruginosa NIER10015, M. ichtyoblabe BIER10025, BIER10040, M. novacekii NIER10029, M. wesenbergii NIER10068) were tested with four rRNA - targeted oligonucleotide probes labelled with horseradish peroxidase (HRP) and specific for cyanobacteria. Non- fluorescent detection of hybridization signal was used. The hybridization with artificial mixture of cyanobacteria have shown the possibility to use 2 species-specific probes in duplicate hybridization and detection with different colored substrates.

  • PDF

Enzyme Immunoassay for Human Chorionic Gonadotropin Using Monoclonal Antibodies (단일크론성 항체를 이용한 융모막 성선자극 호르몬의 효소 면역측정법)

  • 차상훈;김희주;김원배;양중익
    • YAKHAK HOEJI
    • /
    • v.31 no.2
    • /
    • pp.64-69
    • /
    • 1987
  • Monoclonal antibodies against human chorionic gonadotropin (hCG) were prepared and characterized by examining isotype, epitope binding, cross reactivity and affinity constants. And a sandwich type enzyme immunoassay for native hCG was developed with solid phase monoclonal antibody against the conformational determinant expressed only on native hCG and horseradish peroxidase conjugated monoclonal antibody against the $\beta$-subunit of hCG. The assay was sensitive to 1 mIU hCG/ml and shown a linear response up to 200 mIU hCG/ml. The cross reactivity for luteinizing hormone and $\beta$-subunit of hCG were 1% and 0.18%, respectively.

  • PDF

Development of a One-step Two-site Enzyme Immunoassay for Measuring Human Alpha-fetoprotein by Eliminating Hook-effect

  • Kim, Se-Ho
    • BMB Reports
    • /
    • v.34 no.1
    • /
    • pp.47-50
    • /
    • 2001
  • A one-step, two-site enzyme immunoassay was developed for measuring human alpha-fetoprotein (AFP) in serum and amniotic fluid using monoclonal antibodies (McAb) by eliminating the high-dose hook effect. Three McAbs that recognize different epitopes were selected among 16 different clones on the basis of epitope mapping, two for immobilization and one for horseradish peroxidase conjugation. This one-step immunoassay system is more convenient and rapid compared to a conventional two-step sandwich immunoassay system. It did not exhibit the hook effect to around 2.7 mg/ml of AFP, which is probably one of the highest concentrations of AFP in the serum. The dose-response curve of the system was linear to 500 mg/ml of AFP and the system could differentiate as low as 1 mg/ml of AFP The intra- and inter-assay variations were in an acceptable range; 95~104% and 97~105% respectively Its correlation with other commercial systems was around 95%.

  • PDF

Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Clostridium botulinum Type F Toxin (Clostridium botulinum Type F Toxin의 면역학적 효소방법에 의한 검출에 관한 연구)

  • Lee, Jeong-Kug;K. H. Yang
    • Microbiology and Biotechnology Letters
    • /
    • v.10 no.3
    • /
    • pp.205-209
    • /
    • 1982
  • The enzyme-linked immunosorbent assay using the so-called "double-sandwich"technique was applied to determine Clostridium botulinum type F toxin. Polystyrene tubes were coated with horse anti-type F toxin serum and then toxin sample was added. The tubes were subsequently treated with rabbit anti-type F toxin IgG and sheep anti-rabbit serum IgG-horseradish peroxidase conjugate. By this technique, about 10 mouse intraperitoneal 50% lethal doses (ip LD/50/) of type F toxin could be detected. Low back-ground reading was achieved with the use of phosphate-buffered saline containing 0.05% Tween 20 and 1% bovine serum albumin as diluents of rabbit IgG and conjugate. Addition of EDTA in the diluents of toxin increased ELISA extinction value significantly. No cross-reaction was observed with botulinum type A and B toxin, but type E toxin gave sleight cross-reaction.

  • PDF

A Biotin-avidin Labeled Enzyme Immunoassay for the Quantitation of Serum TSH Using Protein-layered Solid Phase

  • Choi, Myung-Ja;Song, Eun-Young;Chung, Tai-Wha
    • Archives of Pharmacal Research
    • /
    • v.21 no.3
    • /
    • pp.231-235
    • /
    • 1998
  • A sensitive enzyme immunoassay for serum TSH has been developed utilizing the tight binding between biotin and avidin, and three layered protein polystyrene beads as solid phase. To increase binding capacity of TSH and sensitivity of the assay, the polystyrene beads were coated sequentially with mouse immunoglobulin as first layer, rabbit antimouse immunoglobulin as second layer and monoclonal anti-TSH as third layer. A serum sample was incubated simultaneously with a monoclonal anti-TSH immobilized polystyrene beads and a second monoclonal anti-TSH covalently attached to biotin. After washing, the antibody bound serum TSH-anti-TSH-biotin complex is reacted with horseradish peroxidase (HRP)-labeled avidin. Following second wash, the bound HRP activity was measured calorimetrically. Reproducible results were obtained within 4 hours for serum TSH in the range between $0{\mu}\textrm{IU}$ml and ${50}{\mu}\textrm{IU}$ml with detection limit of $0.1{\mu}\textrm{IU}$ per test.

  • PDF

Tri-enzyme modified electrochemical biosensor for paracetamol detection (파라세타몰 검출을 위한 전기화학적 다중효소 바이오센서)

  • Park, Deog-Su;Shim, Yoon-Bo;Chang, Seung-Cheol
    • Journal of Sensor Science and Technology
    • /
    • v.17 no.1
    • /
    • pp.29-34
    • /
    • 2008
  • A new disposable amperometric tri-enzyme biosensor for the detection of paracetamol has been developed. The paracetamol sensors developed uses horseradish peroxidase modified screen-printed carbon electrodes (HRP-SPCEs) coupled with immobilized enzymes, tyrosinase and aryl acylamidase, prepared using a poly (vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ) matrix. Optimization of the experimental parameters has been performed and the paracetamol biosensor showed detection limit for paracetamol is as low as $100{\mu}M$ and the sensitivity of the sensor is $1.46nA{\mu}M^{-1}cm^{-2}$.

Detection of luminol chemiluminescence using photodiode (화학 발광 검출을 위한 포토다이오드 제작)

  • Kim, Hyung-Il;Kang, Chul-Goo;Kang, Moon-Sik;Park, Jung-Ho;Min, Nam-Ki
    • Proceedings of the KIEE Conference
    • /
    • 2002.07c
    • /
    • pp.1552-1554
    • /
    • 2002
  • 본 논문에서는 실리콘을 기초로 한 포토다이오드를 제작하여 현재 바이오센서에서 이용되고 있는 luminol 화학 발광을 전기적으로 검출하였다. 우선, 실리콘 웨이퍼에 이온주입을 통해 p-n 접합을 형성하고 Al 전극을 형성시켜 포토다이오드를 제작하였다. PDMS을 passivation 막으로 사용하여 horseradish peroxidase(HRP) 농도 변화에 따른 발광의 최적조건과 luminol, HRP가 섞인 용액에 서로 다른 양의 과산화수소를 넣어 발광이 최대로 발생하는 혼합비를 측정하였다. 최종적으로 2mM luminol, 25U HRP 조건 하에서 과산화수소 농도에 따른 화학 발광을 측정하였으며, $10{\mu}M$부터 $500{\mu}M$ 범위의 과산화수소 농도 변화에 따른 포토다이오드의 감도는 단위 면적당 23.429pA/ ${\mu}M$이다.

  • PDF

Studies on Development of Microplate-EIA for the Determination of Serum Progesterone (혈청 Progesterone 측정을 위한 효소면역분석법 개발에 관한 연구)

  • 김정우;이욱연
    • Korean Journal of Animal Reproduction
    • /
    • v.17 no.4
    • /
    • pp.347-356
    • /
    • 1994
  • A simpled and sensitive microplate enzyme immunoassay(EIA) was developed for the determination of progesterone concentration in serum, based on progesterone monoclonal antibody as anti-progesterone, horseradish peroxidase(HRP) as enzyme-label and tetramethylbenzidine(TMB) as substrate. The assay has a sensitivity of 5 pg-120pg/well and intra- and inter-assay coefficients of variation for progesterone standard curve (1.0ng~10.0ng/ml) were ranged 2.5~9.9% and 1.7.8.0%, respectively, determination coefficient of the regressio equation of our standard curve(R2=0.990$\pm$0.007) were high, and this is the same level as that of commercial kit(Hormonost Bio-Lab, Germany, R2=0.98~0.99). The progesterone concentration of serum determined by both kits (Work & Bio-Lab) were significantly correlated (r=0.95, P<0.01) although a little higher value were resulted in our kit than that of commercial kit. It generally is these results indicated that the microplate-EIA can be cused for the determination of progesterone in serum, as well as, for the determination of the early pregnancy diagnosis.

  • PDF

Experimental Study on Development of ELISA Method for the Detection of Sulfamethazine Residues (잔류 Sulfamethazine 검출용 ELISA 개발에 관한 실험적 연구)

  • 임윤규;김성희
    • Journal of Food Hygiene and Safety
    • /
    • v.10 no.4
    • /
    • pp.213-217
    • /
    • 1995
  • A screening method has been developed for detecting sulfamethazine(SMZ) contamination of meat or feeds by using horseradish peroxidase (HRP) labeled protein A (Prot AHRP)and an indirect competitve enzyme-linked immunosorbent assay(ELISA). The assay is based on competitve binding of guinea pig anti-SMZ with SMZ in smaple and SMZ-gelatin conjugate(SMZ.GEL). Percent binding (B.Bo$\times$100) was calculated from the absorbance in the absence (B0) and presence (B) of SMZ. By the sandard curve prepared by plotting log(SMZ) vs percent binding of each known reference solution, the detection limit was 1.0ppb or less. Cross reacton with sulfadimethoxine, sulfaguaniding, sulfamerazine, sulfamthoxpyridazine, sulfanilamide, sulfisomidine and sufisoxazole were not observed. But sulfamerazine crossreacted in the test. The EC-50 value (concentration causing 50% inhibition of color development compared with blank) of sulfamerazine was 2.0 ppm. Further quality control will make the ELISA system ideal for the detection of SMZ in meat or feeds.

  • PDF