• Animal cells and systems
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• v.7 no.1
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• pp.89-92
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• 2003

Immunoassays have become indispensable tools and achieved great importance in scientific and medical research. However, typical immunoassays are time-consuming and use complex, multi-step procedures. In this study, we introduce a new immunoassay system for the quantification of several analytes at a time without any washing steps. It is comprised of a detector solution with fluorescence-labeled antibodies and a test strip with immobilized capture antibodies. Using a micro-array scanner, the antigen-antibody complex was quantitatively determined by measuring the intensities of fluorescence on the capture lines or dots of nitrocellulose membrane. This method demonstrated its rapid quantitative determination of analytes without many processing steps as well as specific identification of multiple analytes in biological specimens.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.421-421
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• 2005

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1152-1161
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• 2007

An immunochromatography (ICG) strip test based on a monoclonal antibody for the rapid detection of L. monocytogenes in meat and processed-meat samples was developed in this study. A monoclonal antibody (MAb) specific to L. monocytogenes was produced from cloned hybridoma cells (FKLM-3B12-37) and used to develop an ICG strip test. The antibody showed a stronger binding to L. monocytogenes than other Listeria species, and a weak cross-reaction to S. aureus based on an ELISA. The detection limit of the ICG strip test was $10^5\;cell/ml$. In total, 116 meat and processed-meat samples were collected and analyzed using both the ICG strip test and a PCR. The ICG strip test and PCR indicated L. monocytogenes contamination in 34 and 27 meat samples, respectively. The 7 meat samples not identified as L. monocytogenes positive by the PCR were also tested using an API kit and found to be contaminated by Listeria species. In conclusion, the ICG strip test results agreed well with those obtained using the PCR and API kit. Thus, the developed ICG has potential use as a primary screening tool for L. monocytogenes in various foods and agricultural products, generating results within 20 min without complicated steps.

• Journal of environmental and Sanitary engineering
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• v.1 no.1 s.1
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• pp.69-79
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• 1986

This studies were carried out to investigate the high infection sites from various specimens, cultural isolation on the susceptible media and specific antibody titres of M. pulmonis in experimental 310 mice and 330 rats obtained from two breeding facilities. Efficiency of complement faxation test (CF test) for detection of M. pulmonis antibody in mice and rats were compared directly with the diagnostic cultural isolation method. 1. Isolation rates of M. pulmonis among infection sites were about 30% from the oral cavities and $40\%$ from the middle ear of mice. The rates were $100\%$from the nasal cavaties and $90\%$ from the oral cavities of rats. 2. The infection rates were $12\%$ to A group and $16\%$ to B group of mice. The rates in the rats were $55\%$ to A group and $70\%$ to B group. 3. The M. pulmonis antibody titres by CF test were $73\%$ of total 100 mice in serum dilution below 1:5 (< 1:5), and $24\%$ of total samples in antibody titres above 1:5 (> 1:5), but 3 samples were not showed anticomplementary activities. The antibody titres in rats were $35\%$ of 120 rats in below 1:5 (< 1:5), and $61\%$ of total samples in above 1:5 (> 1:5), but the remained were not showed anticomplementary activities.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.546-547
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• 2005

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.425-431
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• 2006

The development of fermentation conditions for the production of C595 diabody fragment (dbFv) in E. coli HB2151 clone has been explored. Investigations were carried out to study the effect of carbon supplements over the expression period, the comparison of C595 dbfv production in synthetic and complex media, the influence of acetic acid upon antibody production, and comparison of one-stage and two-stage processes operated at batch or fed-batch modes in bioreactor. Yeast extract supplied during expression yielded more antibody fragment than any other carbon supply. The synthetic medium presented higher specific productivity (0.066 mg dbFv $g^{-1}$ dry cell weight) when compared to the complex medium (0.044 mg dbFv $g^{-1}$ DCW). The comparison of fermentation strategies demonstrated that (1) one-stage fed-batch fermentation performed higher C595 dbFv production than that operated in batch mode which was significantly affected by acetate concentration; (2) a two-stage batch operation could enhance C595 dbFv production. It was found that a concentration of 12.3 mg $L^{-1}$ broth of C595 dbFv and a cell concentration of 10.8g $L^{-1}$ broth were achieved at the end of two-stage operation in 5-L fermentation.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.663-669
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• 2000

The fur (ferric uptake regulation) expression vector pMON2064 was modified to produce a Fur-fusion expression vector. A kinker site, factor Xa cleavage site, and several restriction endonuclease sites were introduced to facilitate easy cloning and isolating of the fusion protein. The resulting fusion expression vector, pMONSTER, was then used to make fusion expression vector, pMONSTER, was then used to make fusion proteins with $\beta$-galactosidase and the protease of the human immunodeficiency virus type 1 (HIV-1 PR). Strain SW4020 harboring the Fur $\beta$-galactosidase fusion vector produced blue colonies on a 5-bromo-4-chloro-3-indolyl-$\beta$-D-galactoside plate and the resulting 133 kDa fusion protein reacted with an anti-Fur antibody. The strain harboring the Fur-HIV-1 PR fusion vector produced a 29 kDa fusion protein, which also reacted with an anti-Fur antibody. The Fur-HIV-1 PR fusion protein was purified by a single column application that was designed to isolate the Fur protein. The purified Fur-HIV-1 PR fusion protein digested with factor Xa cleaved a recombinant Gag protein to release smaller fragments, including a p24 capsid protein. The Fur-HIV-1 PR fusion protein itself did not exhibit any proteolytic activity.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.21
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• pp.9375-9378
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• 2014

Background: A piezoelectric immunosensor for early cervical cancer detection was developed involving short analyis time and less invasive technique for $p16^{INK4a}$, a protein that has been linked to cervical cancer. Materials and Methods: $5{\mu}L$ of 5.0 mg/mL $p16^{INK4a}$ antibody and then supernatant from different clinical samples from West China Second University Hospital (Sichuan, China) were dripped on the center of the AT-cut crystal through a micro-injector. Absorption of the $p16^{INK4a}$ by antibody caused a shift in the resonant frequency of the immunosensor, and the resonant frequency was correlated to the amount of the $p16^{INK4a}$ in the supernatant. Results: The greater severity of lesion grading, the greater the expression level of $p16^{INK4a}$. Conclusion: Degree of cervical cancer lesion development could be determined by detected amount of $p16^{INK4a}$ in different clinical samples.

• Journal of Sensor Science and Technology
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• v.20 no.6
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• pp.400-405
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• 2011

This study introduced the development of a strip type disposable enzyme-linked immunosensor platform for the detection of IgG. Strips of the strip sensor were fabricated by using commercial nitrocellulose filter membranes and a housing holder for the strips was manufactured by using a standard injection molding process for a plastic material. An IgG-urease conjugate was prepared and used for the competitive immune-binding with sample IgG. From the enzymatic reaction between the conjugated urease and urea added, ammonia was generated and caused a localized alkaline pH change on the immobilized antibody band which was coated onto the sensor strips. This pH increase subsequently caused a color change of the antibody band in the presence of a pH indicator, phenol red. Used in conjunction with a competitive immunoassay format, the intensity of the color produced is directly linked with the concentration of target analyte, IgG, and specific measurement of IgG in a lateral flow immunoassay format was achieved over the range 100 ppb to 2000 ppb IgG.

• Journal of Biosystems Engineering
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• v.34 no.4
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• pp.254-259
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• 2009

Conventional methods for pathogen detection and identification are labor-intensive and take several days to complete. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella typhimurium. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on either avidin-biotin binding or self assembled monolayer (SAM) on the surface of the IME to form an active sensing layer. To evaluate effect of antibody immobilization methods on sensitivity of the sensor, detection limit of the biosensor was analyzed with Salmonella samples innoculated in phosphate buffered saline (PBS) or food extract. The impedimetric biosensor based on SAM immobilization method produced better detection limit. The biosensor could detect 107 CFU/mL of Salmonella in pork meat extract. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.