• Biomolecules & Therapeutics
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• v.8 no.2
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• pp.107-112
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• 2000

To investigate the alteration of airway mucin in airway disease patients, immunoassay procedures were employed using monoclonal antibodies HM02 and HM03 (Hybridoma, 18,457-463, 1999). Alteration of mucin release was determined by ELISA and the integrity of mucin was determined by Western blot. In ELISA, it was found that mucin release increased from pneumonia, chronic cough, bronchiectasis, eosinophilic pneumonia, lung cancer and bronchial asthma patients. In Western blot, the increase in immunoreactivity was observed in case of pneumonia, chronic cough, bronchiectasis and bronchial asthma. In bronchial asthma, there was no obvious degradation of mucin while in other diseases, varying degree of mucin degradation was observed. The data from the present study implicate that HMO2 and HM03 are suitable for the immunological analysis of mucin in airway disease patients. The role of increased mucin release and varying degree of mucin degradation on airway diseases should be further investigated in the future.

• Korean Journal of Veterinary Research
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• v.56 no.3
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• pp.189-192
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• 2016

The virus neutralization (VN) test was used to determine potency of the infectious bronchitis (IB) vaccine. The results of VN, hemagglutination inhibition (HI), and enzyme-linked immunosorbent assay (ELISA) were compared with those of the IBV M41. The $r^2$ values between VN and HI titers and the ELISA antibody titer were 0.8782 and 0.0336, respectively, indicating a high correlation between VN and HI, but not VN and ELISA. The Cohen's kappa coefficient between the VN titer of 2 $log_{10}$ and HI titer of 5 $log_2$ was 0.909. Our results showed that VN could be replaced with HI for testing the potency of IBV M41.

• The Korean Journal of Food And Nutrition
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• v.19 no.3
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• pp.296-300
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• 2006

Immunoblotting and competitive indirect enzyme-liked immunosorbent assay(Ci-ELISA) was used for detection of ${\beta}$-lactoglobulin(BLG) in dairy products, such as milk, dried milk and fermented milk. In immunoblotting, human IgE weakly recognized proteins of fermented milk, but still responded to dried milk even though become weak. Rabbit polyclonal antibody to BLG, used as a model of antigen, and milk allergic patients' IgE was used in the ELISA. Reactivities of Abs were the highest in market milk. BLG in fermented milk was detected in a low content. This result indicates the fermented milk have the lowest BLG content and could be used as hypo-allergenic food for milk-allergic individual.

• Korean Journal of Veterinary Service
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• v.37 no.4
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• pp.241-246
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• 2014

Q fever is a zoonotic disease caused by the obligate intracellular bacterium Coxiella (C.) burnetii and affects wild and domestic animals worldwide. The aim of this study was to evaluate the seroprevalence of C. burnetii in native Korean goat (Capra hircus coreanae) in Gyeongbuk province, Korea, using ELISA. A total of 256 goat blood samples from 56 farms in Gyeongbuk province were collected between May 2012 and March 2013. Among them, 22 (8.6%) samples from 10 (17.9%) farms were seropositive for C. burnetii by ELISA. According to regional analysis, the seroprevalences among goat farms in eastern, western, southern, and northern areas of Gyeongbuk province were 0%, 18.2%, 36.8%, and 6.3%, respectively, showing the highest seroprevalence in the southern region. Among 22 counties in Gyeongbuk province, 10 (45.5%) counties had one or more farms positive to C. burnetii antibody. Accordingly, the seroprevalence of C. burnetii in high-risk humans and animals are constantly demanded by regional investigation.

• Bulletin of the Korean Chemical Society
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• v.26 no.6
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• pp.939-942
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• 2005

For the detection of cyanobacterial toxin, an Enzyme-linked immunosorbent assay (ELISA) was integrated into a PDMS microchip. The conjugates of microcystin-LR (MCLR) and keyhole limpet hemocyanin (KLH) were adsorbed on the surface of polystyrene beads and these MCLR-KLH polystyrene beads were introduced into a microchamber. MCLR on the surface of polystyrene beads reacted with horseradish peroxides (HRP) conjugated anti-MCLR monoclonal antibody (mAb) which had a competitive reaction with MCLR in water sample. After the enzyme substrate 3,3,5,5-tetramethyl benzidine (TMB) was injected into the chamber and catalyzed by HRP, the color change was detected with a liquid-cord waveguide. This integration shortened the conventional ELISA analysis time from several hours to about 30 min with only 4.2 $\mu$L MCLR sample consuming which was useful for the environmental analysis. More over, troublesome operations required for ELISA could be replaced by simple operations. The microchip based detection system showed a good sensitivity of 0.05 $\mu$g/L and maintained good reliability through its quantitative range with low coefficients of variation (2.5-10.5%).

• Bulletin of the Korean Chemical Society
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• v.30 no.1
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• pp.77-82
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• 2009

Thrombin activatable fibrinolysis inhibitor (TAFI) also known as plasma procarboxypeptidase B or U is a 60 kD glycoprotein, which is the major modulator of fibrinolysis in plasma. TAFI is a proenzyme, which is activated by proteolytic cleavage to an active carboxypeptidase B-like enzyme (TAFIa, 35.8 kD) by thrombin/thrombomodulin and plasmin. Modulation of fibrinolysis occurs when TAFIa enzymatically removes C-terminal lysine residues of partially degraded fibrin, thereby inhibiting the stimulation of tissue plasminogen activator (t-PA) modulated plasminogen activation. TAFIa undergoes a rapid conformational change at $37{^{\circ}C}$ to an inactive isoform called TAFIai. Potato tuber carboxypetidase inhibitor (PTCI) was shown to specifically bind to TAFIa as well as TAFIai. In this study, a novel immunoassay TAFIa/ai ELISA was used for quantitation of the two TAFI activation isoforms TAFIa and TAFIai. The ELISA utilizes PTCI as the capture agent and a double antibody sandwich technique for the detection. Low levels of TAFIa/ai antigen levels were detected in normal plasma and elevated levels were found in hemophilia A plasmas. TAFIa/ai antigen represents a novel marker to monitor fibrinolysis and TAFIa/ai ELISA may be a valuable assay for studying the role of TAFI in normal hemostasis and in pathological conditions.

• Parasites, Hosts and Diseases
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• v.40 no.4
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• pp.177-180
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• 2002

Gangweon-do is known to be highly endemic area of sparganosis more than other provinces in Korea. A seroepidmiologic examination for the detection of anti-Spirometra erinacei plerocercoid IgG in serum was carried out in normal inhabitants in Hongcheon-gun, Gangweon-do. Sere were tested by enzyme-linked immunosrobent assay (ELISA) for the anti-sparganum antibodies. Positive rate for anti-sparganum antibody in 719 adults was 3.3%. Data of the questionnaire for 24 ELISA positive inhabitants revealed that 20 had a history of eating raw meat of snakes, 24 had a history of eating frogs, and 24 had a history of drinking stream water. Two positive cases had a past history of sparganosis. Two positive cases showed current symptoms of sparganosis. The data revealed that ELISA would be useful to find infected cases among normal inhabitants at sparganosis-endemic areas.

• Parasites, Hosts and Diseases
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• v.26 no.1
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• pp.1-8
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• 1988

Anti-Clonorchis IgG antibody levels in serum were observed by ELISA in 129 egg positive cases and in 25 controls. The antibody levels were 0.063 to 1.216 (0.325±0.202) in clonorchiasis cases and 0.078 to 0.670 (0.255±0.133) in controls. The difference was statistically significant. However, serological diagnosis of clonorchiasis was not satisfactory in lightly infected cases because of low levels of specific lgG antibody. The antibody levels were well correlated with EPG. Changes of the IgG antibody levels were not signiscant 12∼14 days, 4 weeks and 8∼9 weeks after praziquantel treatment. Seven and 13 months after treatment, the IgG antibody levels were lowered significantly. The period for serologically negative conversion after prasiquantel treatment was between 9 weeks and 7 months in human clonorchiasls.

• Journal of Life Science
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• v.28 no.12
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• pp.1416-1423
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• 2018

With strong biotin binding affinity ($K_D=10^{-14}M$), the tetrameric feature of streptavidin could be used to increase the antigen binding activity of a camel heavy chain (VHH) antibody through their fusion, here stained with biotinylated horseradish peroxidase and subsequent immunoassays ELISA and Western blot analysis. For this application, we cloned the streptavidin gene amplified from the Streptomyces avidinii chromosome by PCR, and this was fused to the gene of the 8B9 VHH antibody which is specific to green fluorescent protein (GFP) antigens. To express a soluble fusion protein in Escherichia coli, we used the pUC119 plasmid-based expression system which uses the lacZ promoter for induction by IPTG, the pelB leader sequence at the N-terminus for secretion into the periplasmic space, and six polyhistidine tags at the C-terminus for purification of the expressed proteins using an $Ni^+$-NTA-agarose column. Although streptavidin is toxic to E. coli because of its strong biotin binding property, this soluble fusion protein was expressed successfully. In SDS-PAGE, the size of the purified fusion protein was 122.4 kDa in its native condition and 30.6 kDa once denatured by boiling, suggesting the tetramerization of the monomeric subunit by non-covalent association through the streptavidin moiety fusing to the 8B9 VHH antibody. In addition, this fusion protein showed biotin binding activity similar to streptavidin as well as GFP antigen binding activity through both ELISA and Western blot analysis. In conclusion, the protein resulting from the fusion of an 8B9 VHH antibody with streptavidin was successfully expressed and purified as a soluble tetramer in E. coli; it showed both biotin and GFP antigen binding activity suggesting the possible production of a tetrameric and bifunctional VHH antibody.

• The Journal of the Korean Society for Microbiology
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• v.20 no.1
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• pp.91-102
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• 1985

The advantages of enzyme-linked immunosorbent assay(ELISA) are its senstivity and simplicity in detecting IgG, IgM and IgA antibody. To apply ELISA to diagnosis of typhoid fever, antigen such as lipopolysaccharide of Salmonella typhi or killed whole cell must be coated on solid phase. It is easy to coat lipopolysaccharide on ELISA plate but troublesome to purify it. As it is easy to obtain the killed whole cells, the development of the appropriate method by which those antigens of S. typhi are optimally coated on solid phase is needed. To establish the appropriate method, carbonate buffer, methanol or poly-L-lysine was applied as binding substance on polystyrene or polyvinylchloride plate as solid phase when the killed whole cell antigens of S. typhi varided as follows: $10^6$, $10^7$, $10^8$ and $10^9\;cell/ml$. The criteria of the optimal method were determined as follows: 1. The optical density of positive sera is above 1.0(0.6 in IgM) at 1:10 serum dilution and is 0.3(0.2 in IgM) higher than that of negative sera: 2. The O.D. of sera is flat or lowering according to serum dilution: 3. It must be that the O.D. of negative sera is lower than 0.2 at the point of serum dilution where the O.D. of positive sera is higher than 1.0(0.5 in IgM). The results obtained were summarized as follows: 1. The methods which fitted the above criteria were to use poly-L-lysine as binding substance, polyvinylchloride plate as solid phase and $10^7\;cell/ml$ as antigen concentration of S. typhi(poly-L-lysine/polyvinylchloride/$10^7$) and poly-L-lysine/polyvinylchloride/$10^8$ in detecting IgG antibody, methanol/polystyrene/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgM and carbonate buffer/polystyrene/$10^8$, carbonate buffer/polystyrene/$10^9$, methanol/polystyrene/$10^8$, methanol/polyvinylchloride/$10^8$, methanol/polyvinylchloride/$10^9$, poly-L-lysine/polyvinylchloride/$10^8$ and poly-L-lysine/polyvinylchloride/$10^9$ in IgA. 2. The coaling method using poly-L-lysine, polyvinylchloride plate and $10^8\;cell/ml$ was best to assay IgG, IgM and IgA antibody all in one. By this method, to assay the each immunoglobulin calss with an appropriate fixed serum dilution, 1:320 dilution was best.