• Journal of Biomedical Engineering Research
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• v.30 no.2
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• pp.122-128
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• 2009

Cancer serum biomarkers have advanced our ability to more accurately predict tumor classification, prognostic/metastatic potential, and response potential to novel chemotherapies. Serum amyloid A (SAA) and Vascular endothelial growth factor (VEGF) have potential utility as a serum biomarker for lung cancer. Quantum dots, nanometer-sized crystals, have a high quantum yield, sensitivity, and pronounced photostability. The properties of quantum dots can be efficiently applied to the detection of serum biomarkers in immunoassays as fluorescent probe. We used quantum dots as fluorescent probes in immunoassays and attempted to detect serum amyloid A and vascular endothelial growth factor as serum biomarkers of lung cancer. This fluorescence immunoassay based on the properties of quantum dots is applicable to the detection of serum biomarkers for lung cancer. The fluorescence immunoassay with quantum dots should allow the efficient and specific detection of serum amyloid A (SAA) for the possible diagnosis of lung cancer.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1216-1221
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• 2006

A protein array was fabricated for a miniaturized immunoassay using microcontact printing ($\mu$CP). A polydimethylsiloxane (PDMS) stamp with a 5 $\mu$m$\times$5 /$\mu$m dimension was molded from a silicon master developed by photolithography. Under optimal fabrication conditions, including the baking, incubation, and exposure time, a silicon master was successfully fabricated with a definite aspect ratio. An antibody fragment was utilized as the ink for the $\mu$CP, and transferred to an Au substrate because of the Au-thiol (-SH) interaction. The immobilization and antibody-antigen interaction were investigated with fluorescence microscopy. When human serum albumin (HSA) was applied to the protein array fabricated with an antibody against HSA, the detection limit was 100 pg/ml of HSA when using a secondary antibody labeled with a fluorescence tag. The fabricated protein array maintained its activity for 14 days.

• Toxicological Research
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• v.7 no.1
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• pp.29-35
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• 1991

Barbiturates are commonly abused tranquilizer and a rapid method to determine these drugs in biological samples is needed. In this study, was screened barbiturates in urine specimens by the fluorescence polarization immunoassay method(FPIA) and the positive samples were confirmed and identified by the more definitive GC/MS method. Fifteen positive smples which have barbiturate values higher than 0.5 ng/ml were analyzed by the GC/MS method. Eight samples were identified as phenobarbital and five samples were identified as crotilbarbitone.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.979-985
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• 2001

A fluorometric detection technique for HDL (High Density Lipoprotein) and LDL (Low Density Lipoprotein) was developed for application in a fiber-optic immunosensor using a protein A Langmuir-Blodgget (LB) film. For the fluorescence immunoassay, antibodies specific to HDL or LDL were imobilied on the protein A LB film, and a fluorescence amplification method was developed to overcome their weak fluorescence. The deposition of protein A using the LB technique was monitored using a surface pressure-are $({\pi}-A)$ curve, and the antibody immobilization of the protein A LB film was experimentally verified. The immobilized antibody was used to separate only HDL and LDL from a sample, then the fluorescence of he separated HDL or LDL was amplified. The amount of LDL or HDL was measured using the developed fiber optic fluorescence detection system. The optical properties resulting from the reaction of HDL or LDL with o-phtaldialdehyde, detection range, response time, and stability of the immunoassay were all investigated. The respective detection ranges for HDL and LDL were sufficient to diagnose the risk of coronary heart disease. The amplification step increased the sensitivity, while selective separation using the immobilized antibody led to linearity in the sensor signal. The regeneration of the antibody-immobilized substrate could produce a stable and reproducible immunosensor.

• 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.

• Biomedical Science Letters
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• v.22 no.4
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• pp.160-166
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• 2016

To provide a basis for a homogeneous fluorescence resonance energy transfer (FRET) immunoassay for celiac disease, we carried out a FRET experiment using guinea pig tissue transglutaminase (tTG) and antibodies to tTG (anti-tTG) purified from rat serum. Fluorescein was utilized as the probe, and a nonfluorescent dye, QSY 7 served as the quencher. We labeled anti-tTG and tTG with fluorescein isothiocyanate and QSY 7 succinimidyl ester, respectively. Fluorescein-labeled anti-tTG was the donor, and QSY 7-labeled tTG was the acceptor of the FRET experiment. When we titrated fluorescein-labeled anti-tTG with QSY 7-labeled tTG, we observed a large decrease in the steady-state fluorescence intensity, which was due to strong FRET from fluorescein-labeled anti-tTG to QSY 7-labeled tTG. Using time-resolved fluorescence spectroscopy, we could also observe a decrease in the fluorescence lifetime, which confirms the steady-state data. We expect that these results might be useful in the development of a novel fluorescence immunoassay for an easy screening and follow-up of celiac patients.

• Korean Journal of Veterinary Research
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• v.48 no.2
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• pp.175-179
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• 2008

Parallux, a solid-phase fluorescence immunoassay (SPFIA) developed for detection antibiotics residue in milk, was applied for analysis of antibiotics in muscle tissue of olive flounder (Paralichthys olivaceus), rockfish (Sebastes schlegeli), and red sea bream (Pagrus major). Fishes were dipped in neomycin 140 mg/ton water, the recommended therapeutic dose, for 24 h. Muscle samples were obtained on 1st, 2nd, 3rd, 4th and 5th day after drug treatment. The concentration of neomycin in muscle was determined using an internal standard (100 ppb as neomycin). The absorbance ratio of sample to internal standard (S/C) was employed as an index to determine the muscle residues in fishes. To investigate the recovery rate, the standard solutions were added to muscle samples to give final concentrations in muscle of 0.2 and 0.5 mg/ml. The recovery rates of all spiked samples were > 85% of the spiked value. Neomycin was detected in muscles of fishes treated after the 1st day of withdrawal period. On the 2nd day after drug treatment, all muscle samples showed negative reaction (S/C ration ${\leq}$ 1.0). The present study showed that the SPFIA can be applied for predicting residues of neomycin in muscle tissues of farmed fishes.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.291-294
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• 2006

$Parallux^{TM}$, a solid-phase fluorescence immunoassay(SPFIA) developed for antibiotics residue detection in milk, was applied for analysis of fish muscle. The recommended therapeutic dose of ampicillin(100 mg/kg body weight, withdrawal period 7 days) was orally administered to a group of 25 olive flounders(Paralichthys olivaceus) for consecutive five days. Muscle was sampled after drug treatment 1st, 2nd, 3rd, 4th and 5th day. The concentration of ampicillin in muscle, determined by SPFIA, was compared with that of internal standard(10 ppb as ampicillin). The absorbance ratio of sample to internal standard(Bs/Bo) was employed as an index to determine the muscle residue in olive flounder. To investigate the recovery rate, the standard solutions were added to muscle samples to give final concentrations in muscle of 10 and 50 ng/ml. The recovery rates of all spiked samples were > 89% of the spiked value. Ampicillin was detected in muscle of fishes treated until the 3rd day of withdrawal period. The present study showed that the SPFIA can be easily adopted in predicting tissue residues for ampicillin in farmed fishes.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.28 no.9
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• pp.1429-1434
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• 2004

Immunoassay is one of the important analytical methods for clinical diagnoses and biochemical studies, but needs a long time, troublesome procedures and expensive reagents. In this study, therefore, we propose the micro filter chip with microbeads for immunoassay, which has pillar structures. The advantage of the proposed micro filter chip is to use simple fabrication process and cheap materials. The mold was made by the photolithography technique with Si wafer and negative photoresist SU-8. The replica was made of PDMS, bonded on the pyrex glass. The micro filter chip consists of inlet channel, filter chamber and outlet channel. HBV (Hepatitius B virus) monoclonal antibody (Ag1) labeled with biotin were immobilized onto streptavidin coated beads of 30∼50 $\mu$m size. Fluorescein isothiocyanate (FITC)-labeled HBV monoclonal antibody (Ag8) was used to detect HBsAg (Hebatitis B virus surface Antigen), and fluorescence intensity was monitored by epi-fluorescence microscope. In this study, the immune response of less than 30 min was obtained with with the use of 100 $m\ell$ of sample.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.325-333
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• 1993

A competitive type immunoassay method for 17$\beta$-estradiol(E2) based on the idiotypic anti-idiotypic antibody and time-resolved fluorescence is described. The anti-idiotypic antibody(Ab2) produced to E2 binding site of the primary idiotype antibody (Ab1) was labelled with europium and was allowed to compete with E2 standards or serum sample for the binding sites of Ab1 which was bound to 2nd antibody captured ontothe surface of microtitre plates. Fluorescence measured by time-resolved fluorometer was inversely proportional to the concentration of E2 over the range 5~500pg/well. The sensitivity of the assay was 5pg per well which was compatible with that ofradioimmunoassay using the same Ab1 and 3H-E2 as a tracer. One great advantage of this method described here was to enable antibodies to be labelled instead of haptens, and thus makes it easier to develop sensitive and robust immunoassay systems specially for haptens.