• Title/Summary/Keyword: detection of antibody

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Characteristics of Protein G-modified BioFET

  • Sohn, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.20 no.4
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    • pp.226-229
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    • 2011
  • Label-free detection of biomolecular interactions was performed using BioFET(Biologically sensitive Field-Effect Transistor) and SPR(Surface Plasmon Resonance). Qualitative information on the immobilization of an anti-IgG and antibody-antigen interaction was gained using the SPR analysis system. The BioFET was used to explore the pI value of the protein and to monitor biomolecular interactions which caused an effective charge change at the gate surface resulting in a drain current change. The results show that the BioFET can be a useful monitoring tool for biomolecular interactions and is complimentary to the SPR system.

Surface Plasmon Resonance Immunosensor for Detection of Legionella pneumophila

  • Oh, Byung-Keun;Lee, Woochang;Bae, Young-Min;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.112-116
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    • 2003
  • An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.

Detection of Pathogenic Salmonella Using a Surface Plasmon Resonance Biosensor (표면플라즈몬공명 바이오센서를 이용한 살모넬라 검출)

  • Cho, Han-Keun;Kim, Gi-Young;Kim, Woon-Ho;Sung, Min-Sun
    • Journal of Biosystems Engineering
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    • v.35 no.2
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    • pp.116-123
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    • 2010
  • Rapid detection of foodborne pathogens has been a major challenge for the food industry. Salmonella contamination is well known in all foods including pasteurised milk. The possibility of specific detection of Salmonella Enteritidis by surface plasmon resonance (SPR) biosensor was explored using a commercially available portable SPR sensor. Self assembly technique was adopted to immobilize anti-Salmonella antibodies on the gold sensing surface of the SPR sensor. The concentration of polyclonal antibody for use in the SPR biosensor was chosen to 1.0 mg/mL. Experiments were conducted at near real-time with results obtained for one SPR biosensor assay within 1 hour. The limit of detection for Salmonella Enteritidis was determined to be $10^6$ CFU/mL in both PBS buffer and milk samples. The assay sensitivity was not significantly affected by milk matrix. Our results showed that it would be possible for employing the SPR biosensor to detect Salmonella Enteritidis in near real-time.

Cell-SELEX Based Identification of an RNA Aptamer for Escherichia coli and Its Use in Various Detection Formats

  • Dua, Pooja;Ren, Shuo;Lee, Sang Wook;Kim, Joon-Ki;Shin, Hye-su;Jeong, OK-Chan;Kim, Soyoun;Lee, Dong-Ki
    • Molecules and Cells
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    • v.39 no.11
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    • pp.807-813
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    • 2016
  • Escherichia coli are important indicator organisms, used routinely for the monitoring of water and food safety. For quick, sensitive and real-time detection of E. coli we developed a 2'F modified RNA aptamer Ec3, by Cell-SELEX. The 31 nucleotide truncated Ec3 demonstrated improved binding and low nano-molar affinity to E. coli. The aptamer developed by us out-performs the commercial antibody and aptamer used for E. coli detection. Ec3(31) aptamer based E. coli detection was done using three different detection formats and the assay sensitivities were determined. Conventional Ec3(31)-biotin-streptavidin magnetic separation could detect E. coli with a limit of detection of $1.3{\times}10^6CFU/ml$. Although, optical analytic technique, biolayer interferometry, did not improve the sensitivity of detection for whole cells, a very significant improvement in the detection was seen with the E. coli cell lysate ($5{\times}10^4CFU/ml$). Finally we developed Electrochemical Impedance Spectroscopy (EIS) gap capacitance biosensor that has detection limits of $2{\times}10^4CFU/mL$ of E. coli cells, without any labeling and signal amplification techniques. We believe that our developed method can step towards more complex and real sample application.

Meaning of Anti Hepatitis C Virus Antibody Detection in Patients Treated with Intravenous Immunoglobulin (인민역 글로불린 제재를 투여받은 환아에서 C형 간염바이러스 항체(anti-HCV) 검출의 의의)

  • Lee, Myong-Gyun;Lee, Young-Hwan;Shin, Son-Moon;Hah, Jeong-Ok
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.306-312
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    • 1993
  • To evaluate the meaning of anti-HCV detection in patients treated with IVIG, serum levels of aspartate aminotranstferase(AST), alanine aminotransterase(ALT), HCV Ab titer were measured after treatment with IVIG in 36 patients diagnised of Kawasaki disease or neonatal sepsis. Also polymerase chain reaction (PCR) for the detection of HCV was done in 8 patients with persistent HCV Ab positivity at 3 months after IVIG treatment. The results were as follows 1) HCV Ab was positive in all 36 patients at 1 week after IVIG treatment, but in only 8 cases it was positive at 3 months after IVIG treatment. 2) AST, ALT were elevated in 9 cases at 1 week after IVIG treatment, but they were normalized in all cases at 3 months after IVIG treatment. 3) PCR for the detection of HCV was done in 8 patients with persistent HCV Ab positivity at 3 months after IVIG treatment, but HCV was not isolated in any cases. These results suggested that detection of anti-HCV was merely transitory phenominon of HCV Ab transmission, did not show any evidence of HCV infection due to HCV transmission.

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A Dipstick-Type Electrochemical Immunosensor for The Detection of The Organophosphorus Insecticide Fenthion

  • Cho, Young-Ae;Cha, Geun-Sig;Lee, Yong-Tae;Lee, Hye-Sung
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.743-746
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    • 2005
  • A dipstick-type immunochemical biosensor for the detection of the organophosphorus insecticide fenthion was developed using a screen-printed electrode system as an amperometric transducer with polyclonal antibodies against fenthion as a bioreceptor. The assay of the biosensor involved competition between the pesticide in the sample and pesticide-glucose oxidase conjugate for binding to the antibody immobilized on the membrane. This was followed by measurement of the activity of the bound enzyme by the supply of the enzyme substrate (glucose) and amperometric determination of the enzyme reaction product ($H_2O_2$). The activity of the bound enzyme was inversely proportional to the concentration of pesticide. The optimized sensor system showed a linear response against the logarithm of the pesticide concentration ranging from $10^{-2}$ to $10^3\;{\mu}g/L$.

Simple Electrochemical Immunosensor for the Detection of Hippuric Acid on the Screen-printed Carbon Electrode Modified Gold Nanoparticles

  • Choi, Young-Bong;Tae, Gun-Sik
    • Journal of the Korean Electrochemical Society
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    • v.14 no.1
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    • pp.44-49
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    • 2011
  • This paper describes an electrochemical immunosensor for simple, fast and quantitative detection of a urinary hippuric acid which is one of major biological indicator in toluene-exposed humans. The feature of this electrochemical system for immunoassay of hippuric acid is based on the direct conjugation of ferrocene to a hippuric acid. With the competition between the ferrocene-hippuric acid complex and hippuric acid for binding to the anti-hippuric acid monoclonal antibody coated onto gold nanoparticles, the electrical signals are turned out to be proportional to urinary hippuric acid in the range of 0.01-10 mg/mL, which is enough to be used for the point-of-care. The proposed electrochemical method could extend its applications to detect a wide range of different small molecules of antigens in the health care area.

Enzyme Immunoassay for Rapid Detection of the Fungicide Iprovalicarb Residues (살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 효소면역분석법)

  • Cho, H.K.;Kyung, K.S.;Lee, E.Y.
    • Journal of Biosystems Engineering
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    • v.31 no.6 s.119
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    • pp.535-540
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    • 2006
  • For a biosensor development, an enzyme-linked immunosorbent assay (ELISA) of the fungicide iprovalicarb was developed by minimizing the processing time. The time for whole incubation process was reduced from 135 minutes to 15 minutes. The concentration of antibody was varied to improve sensitivity. The total processing time was reduced from 2.5 hours to 20 minutes, the final sensitivity ($IC_{50}$ value) of 7.93 ng/mL and the lowest detection limit of 0.045 ng/mL were obtained. This ELISA was applied to potatoes and onions, and the recoveries were in the range of 98.85 $\sim$ 101.20% and 87.97 $\sim$ 102.70%, respectively. Accordingly, this method can be used as basis for a biosensor for rapid monitoring of iprovalicarb residues in crops.

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.