• 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.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.147-151
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• 2004

Antigenicities of ovomucoid (OM) and ovalbumin (OA) in chicken egg white (EW) before and after NaOH, heat, and pretense treatments were examined by competitive indirect enzyme-linked immunosorbent assay (ciELISA), using rabbit anti-OM and-OA antibodies, Enzymatic hydrolysis of EW did not effectively reduce antigenicity of OM, whereas that of OA was decreased to 1/5,000-1/100,000 by treatment of plant-derived or microbial pretenses. Heat treatment below $100^{\circ}C$ for 30min did not decrease antigenicity of OM, whereas that of OA in heated EW increased maximally to 100 times, Antigenicity of OM in EW effectively decreased by NaOH treatment, disappearing at over 1% NaOH, whereas that of OA increased. Additional heat treatment of NaOH-treated EW at $70^{\circ}C$ for 15min slightly reduced antigenicities of OM and OA.

• Journal of Korean Society on Water Environment
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• v.33 no.5
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• pp.538-545
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• 2017

The purpose of this study is to investigate growth response and toxigenicity under various temperature and nutritional conditions, in order to understand the physioecological characteristics of Aphanizomenon flos-aquae, which is a bloom-forming cyanobacterium in the Nakdong River. The strain was inoculated into media under combinations of four temperatures (4, 12, 21, $30^{\circ}C$) and three nutrients (modified CB medium, P-depleted CB medium, N-depleted CB medium) for 28 days. The algae-inhibition tests were performed to assess the potential allelopathic effects of the strains' filtrates on the growth of four algae strains (Microcystis aeruginosa, Aulacoseria ambigua f. spiralis, Aphanizomenon flos-aquae, Scenedesmus obliquus). Toxin production of a strain was measured by Enzyme-Linked ImmunoSolbent Assay (ELISA). The optimal growth temperature (Topt) of strains was $19.9^{\circ}C$ ($18.3-21.2^{\circ}C$), and the temperature range for growth was from $-0.3^{\circ}C$ to $34.3^{\circ}C$. Specific growth rate (${\mu}$) in modified CB medium varied from 0.10 to $0.16day^{-1}$, and the maximum growth rate (${\mu}_{max}$) was $0.17day^{-1}$. Although growth curves under N-existed and N-depleted conditions were almost the same, growth under N-depleted condition was relatively slowed (${\mu}=0.09$ to $0.14day^{-1}$), with a decreased maximum cell density. However, growth under the P-depleted condition was restricted for all temperatures, Two stains of Aphanizomenon flos-aquae were confirmed as not producing toxins, because saxitoxin and cylindrospermopsin were not detected by ELISA. The exudates or filtrates from the Aphanizomenon flos-aquae (DGUC003) resulted in significant inhibition of algal growth on the Aulacoseira ambigua f. spiralis (DGUD001) and Aphanizomenon flos-aquae (DGUC001) (p < 0.01).

• Korean Journal of Veterinary Service
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• v.15 no.1
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• pp.32-45
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• 1992

In order to establish and enzyme-linked immunosorbent assay to ILTV, field virus strain of ILTV was propagated in chorioallantoic membrane of the embryonated eggs. purified and used as antigen. The antisera selected from the field samples and immunized chickens based on serum neutralization test were used as the standard positive and negative sera in all tests. It was found that optimal antigen concentration was $2{\mu}g$ of protein per well and a 1 : 100 dilution of standard serum showed low background optical density with negative serum and high P/N values of positive sera. A 1 : 500 dilution of the rabbit anti-chicken IgG peroxidase conjugate produced a high P/N values and thirty minutes was chosen as suitable time to read the optical density of the enzyme substrate reaction and optical density was consistent during the 16 hours after stopper was treated. When coated antigen was kept on microplate for varying time up to 16 hours at $4^{\circ}C$ or $37^{\circ}C,$ no significant difference was observed between the treatment. The coated antigen could be kept without change of antigenicity for at least one month at $-70^{\circ}C,\; -20^{\circ}C,\; 4^{\circ}C$ and room temperature. When blocking buffer contanining bovine serum albumin was mixed directly with conjugate and serum at 10% level induced higher P/N values compared to blocking antigen coated microplate with the same blocking buffer. The coefficience of correlation between ELISA and SN test was 0.577. When antibody response of chickens, vaccinated with ILTV, was examined by ELISA and SN test, antibody rising and decay pattern between the two test was similar until 11 weeks of age. However 12 weeks onward antibody titer checked on by SN test was slightly lower than that tested by ELISA.

• International Journal of Industrial Entomology and Biomaterials
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• v.29 no.2
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• pp.214-219
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• 2014

We prepared antibodies from insect glycosaminoglycans (GAGs) and assayed the titer. Nine polyclonal antibodies against insect GAGs were raised for development of an ELISA in biological fluids (mice serum). The 3th booster collection of antiserum of BALB/c mice as a primarily antibody was assayed for titer determination by ELISA method. In sandwich ELISA of GAGs derived from Isaria sinclairii or other insects, antiserum from insect GAGs gave satisfactory results for so potent antibody(100: 1~1000:1) raising (manufacturing) agent in range of 10 ng/ml.

• Parasites, Hosts and Diseases
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• v.41 no.3
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• pp.175-177
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• 2003

An ELISA was established to measure bovine IgG directed against the recombinant antigenic determinant of Nc-p43, a major surface antigen of Neospora caninum. In a previous study, two thirds of the C-terminal of the molecule was expressed as a $6{\;}{\times}{\;}His$ tagged protein (Ncp43p) for ELISA using 2/3 of the N-terminal of SAG1 from Toxoplasma gondii as a control (TgSAG1A). Among 852 cattle sera collected from stock farms scattered nation-wide, 103 sera (12.1％) were found to react with Ncp43p positively, but no positive reaction was observed with TgSAG1A. This study shows that Ncp43p could be available as an efficient antigen for the diagnosis of neosporosis in cattle. Furthermore, it together with TgSAG1A, could be useful for the differential diagnosis of N. caninum and T.gondii infections in other mammals.

• Food Science and Preservation
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• v.21 no.5
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• pp.747-756
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• 2014

The objective of this study was to examine the effects of vitamin C on the formation of aflatoxin $B_1$ ($AFB_1$)-DNA adduct and $AFB_1$-induing cellular oxidative damage in rat livers treated with radiation and $AFB_1$. Six-week-old male Sprague-Dawley rats were randomly divided into five groups: the control group, the $AFB_1$-treated group, the group treated with $AFB_1$ and vitamin C, the group treated with X-ray and $AFB_1$, and the group treated with X-ray and $AFB_1$ with vitamin C. On the first day of the experiment, only one dose of X-rays was exposed to the entire liver at 1,500 cGy. Next, vitamin C was injected at 10 mg/kg body weight via intraperitoneal injection, followed an hour later by the administration of 0.4 mg/kg of $AFB_1$ via intraperitoneal injection. These treatments were administered every three days for 15 days. On the 16th day, the animals were sacrificed. The $AFB_1$ contents of the rat sera were determined via indirect competitive ELISA. In the quantitative analysis of $AFB_1$ in the rat sera via ELISA, $5.17{\pm}0.34ng/mL$ of $AFB_1$ was detected in the $AFB_1$-treated groups, but the amount decreased more significantly to $3.23{\pm}0.76ng/mL$ in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. The effect of vitamin C on $AFB_1$-DNA adduct formation was determined via ELISA. The values of $AFB_1$-DNA adduct formation were $9.38{\pm}0.41ng/mL$ in the $AFB_1$-treated groups, but the amount decreased more significantly to $5.28{\pm}0.32ng/mL$ in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. Immunohistochemistry revealed that the accumulation of the $AFB_1$ was not observed in the normal liver tissue (G1). The $AFB_1$-positive materials were observed in the central vein and the portal vein of the liver tissue from the $AFB_1$(G2) treatment or the X-ray and $AFB_1$(G4) co-treatment, but the $AFB_1$-positive materials were observed weakly in the group treated with vitamin C (G3 and G5). These results indicate that vitamin C had ameliorating effects on the $AFB_1$ accumulation of liver tissue.

• Parasites, Hosts and Diseases
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• v.31 no.2
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• pp.141-148
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• 1993

ELISA-inhibition test using Paragonimus westermani specific monoclonal antibody (Mab) was investigated to improve the diagnostic specificity of paragonimiasis. By cell fusion, one hybridoma clone secreting un-n westemanl specific Mab was selected (Pwa-14), which reacted on bands of 28 kDa, 42.5 kDa, 89 kDa and 120.5 kDa. IFA showed Pwa-14 was located at the vitelline follicles. By micro-ELISA, 100% of 22 paragonimiasis cases were found positive, but 5 of 40 clonorchlasls cases (12.5%),3 of 26 cystlcercosis cases (7.7%) showed false positive. None of 10 sparganosis patients or 28 normal controls reacted positively. On the other hand, by ELISA-Inhibition test using a R westermcni specific Mab, 100% of patagonimlasls cases were found positive, and there were no positive in cysticercosis, sparganosis cases or normal controls, except 2 (5.0%) false-positive sera of 40 clonorchiasis cases. The ELISA-Inhlbltlon test using a Mab showed higher specificity in comparison with macro-ELISA for serodlgnosis of human paragonimlasis.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.1
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• pp.235-238
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• 2016

Hepatitis C is an ailment of liver caused by hepatitis C virus (HCV) infection. About 3% of the world population is infected by this virus. HCV infection is a leading reason for liver cirrhosis and therefore a major source of hepatocellular carcinoma. The study focused on the incidence of active HCV infection in blood donors of Mardan district of KPK, Pakistan. A total of 5318 blood donors were inspected for the presence of anti-HCV antibodies and HCV-RNA using ICT (immune-chromatographic test), ELISA and RT-PCR at Mardan Medical Complex (MMC), Mardan. Out of these, 157 (2.95%) were positive by ICT, 60 (1.12%) by ELISA and 56 (1.05%) for HCV-RNA. The frequency of active HCV infectivity amongst the blood donors from district Mardan, KPK Pakistan was 1.05 %. Application of strict measures during blood donor selection and use of proper screening assays such as ELISA in place of ICT devices can give a more accurate picture so that the incidence of this viral infection in HCV negative blood recipients can be reduced.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.6
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• pp.1030-1034
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• 2000

The ovalbumin, a most sensitive egg white protein to irradiation was purified from irradiated hen's eggs. Eggs were irradiated in their shells to 0~7 kGy. To investigate for a practical use in identifying of irradiated eggs, competitive ELISA using ovalbumin was peformed. The binding activity of ovalbumin to anti-ovalbumin IgG was reduced in a dose-dependent manner by irradiating up to 7 kGy, and consider-ably lowered after irradiating at 7 kGy. The concentration of 50% inhibition of ovalbumin to IgG was increased to 1.5~3.7 times in an irradiation dose-dependent relationship. SDS-PAGE of ovalbumin showed that the partial breakdown of ovalbumin was induced by irradiation. The lowering of binding activity was probably due to the partial breakdown of ovalbumin by irradiation. These results demonstrated that the ELISA should be quite useful and effective methods for the identification of irradiated eggs.