• 제목/요약/키워드: ELISA method

검색결과 595건 처리시간 0.031초

파상풍 톡소이드를 함유한 생체분해성 미립구의 특성 (Characteristics of Tetanus Toxoid Loaded in Biodegradable Microparticles)

  • 김지윤;김수남;백선영;이명숙;민홍기;홍성화
    • 약학회지
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    • 제44권4호
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    • pp.293-299
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    • 2000
  • Biodegradable microspheres made from poly-lactide-co-glycolide polymers have been considered as a new delivery system for single-dose vaccine. Purified tetanus toxoid (TT) was encapsulated in poly-lactide(PLA) and poly-lactide-co-glycolide (PLGA) microparticles using a solvent evaporation method in a multiple emulsion system (water-in oil-in water). The morphology of 77-loaded microparticles was spherical and the suface of them was smooth. The particle size was in a range of 2-10. Protein loading efficiency was 68-97.8%. PLGA (85:15) microparticle showed the highest efficiency. Protein release pattern was influenced by polymer molecular weight and composition. The release rate of PLA(Mw 100,000) microsphere was higher than any other microspheres. In consequence of the hydrolysis of PLGA(50:50) microspheres, environmental pH decreased from 7.4 to 5.0. The PLA, PLGA (75:25) and PLGA (85:15) microshperes showed no significant pH change. The antigenicity or n in microshperes was assayed by indirect sandwich ELISA using equine polyclonal tetanus antitoxin for capture antibody and human polyclonal tetanus antitoxin for primary antibody. The antigenicity of TT in PLA (Mw 100,000), PLGA(50:50, Mw 100,000) and PLGA (75:25, Mw 73,300) after 30 days incubation showed 54, 40.9 and 76.7%, respectively.

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Comparison of Some 3-(Substituted-Benzylidene)-1, 3-Dihydro-Indolin Derivatives as Ligands of Tyrosine Kinase Based on Binding Mode Studies and Biological Assay

  • Olgen, Sureyya
    • Archives of Pharmacal Research
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    • 제29권11호
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    • pp.1006-1017
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    • 2006
  • A series of 3-(substituted-benylidene)-1, 3-dihydro- indolin-2-one, 3-(substituted-benylidene)-1, 3-dihydro- indolin-2-thione and 2, 2'-dithiobis 3-(substituted-benylidene)-1, 3-dihydro-indole derivatives was investigated as inhibitor of $p60^{c-Src}$tyrosine kinase by performing receptor docking studies and inhibitory activity toward tyrosine phosphorylation. Some compounds were shown to be docked at the site, where the selective inhibitor PP1 [1-tert-Butyl-3-p-tolyl-1H-pyrazolo[3,4-d]pyrimidine-4-yl-amine] was embedded at the enzyme active site. Evaluation of all compounds for the interactions with the parameters of lowest binding energy levels, capability of hydrogen bond formations and superimposibility on enzyme active site by docking studies, it can be assumed that 3-(substituted-benzylidene)-1, 3-dihydro-indolin-2-one and thione derivatives have better interaction with enzyme active site then 2, 2'-dithiobis 3-(substituted-benzylidene)-1, 3-dihydro indole derivatives. The test results for the inhibitory activity against tyrosine kinase by Elisa method revealed that 3-(substituted-benylidene)-1, 3-dihydro- indolin-2-thione derivatives have more activity then 3-(substituted-benylidene)-1, 3-dihydro- indolin-2-one derivatives.

십전대보탕약침액(十全大補湯藥鍼液)의 사람 피부아세포의 콜라게나제 활성 및 프로콜라겐 합성과 티로시나제 활성에 미치는 영향 (Effects of Sipgeondaebo-tang Pharmacopuncture Extracts on the Collagenase Activity and Procollagen Synthesis in HS68 Human Fibroblasts and Tyrosinase Activity Original Articles)

  • 이세나;김명규;김명희;김형준;조학준;임강현
    • 대한약침학회지
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    • 제14권1호
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    • pp.51-59
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    • 2011
  • Objectives : This study was designed to investigate the collagen metabolism and tyrosinase activity of Sipgeondaebo-tang Pharmacopuncture extracts (SP). Methods : The effect of SP on type I procollagen production and collagenase activity in human normal fibroblasts HS68 after UVB (312 nm) irradiation was measured by ELISA method. The tyrosinase activity after treatment of SP was measured as well. Results : Type I procollagen production was recovered by SP in UVB damaged HS68 cells. The increased collagenase activity after UVB damage was significantly recovered by SP. The tyrosinase activity was significantly reduced as well. However, the L-DOPA oxidation was not changed. Conclusion : SP showed the anti-wrinkle effects and whitening effects in vitro. These results suggest that SP may be a potential pharmacopuncture as an anti-aging pharmacopuncture treatments.

동충하초(冬蟲夏草)추출물이 Ovalbumin으로 유도된 천식(喘息)의 Cytokine에 미치는 영향 (Effects of Cordyceps sinensis Water Extract on the Cytokine in Ovalbumin-induced Asthma Mouse)

  • 정희선;이종록;김상찬
    • 동의생리병리학회지
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    • 제20권4호
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    • pp.973-979
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    • 2006
  • Cordyceps sinensis has been clinically used for the treatment of recovering fatigue, promoting the production of body fluid and qi, resolving sputum and arresting cough. Recent studies showed that Cordyceps sinensis produced immune-modulatory, anticarcinogenic and antioxidative effects. But, there are lack of studies regarding the effects of Cordyceps sinensis on the asthma. So, this study was performed to investigate the oral administrated effects of this herb against the disease. Asthma was induced to Balb/c mouse by i.p. injection and aerosol immunization with ovalbumin, and the change of the eosinophil number in the bronchoalveolar avage fluid(BALF) was observed. Concentrations of Interleukine-4(IL-4), Interleukine-5(IL-5) in BALF and splenocyte were assessed by ELISA, ImmunoglobinG(IgG) and ImmunoglobinE(IgE) from serum were calculated by same method. We found that the effects of Cordyceps sinensis in asthma mouse was implicated in reductions of IL-4, IL-S released from type2 T helper (Th2) cell, and decreases of IgE from plasma cell. These findings suggest that Cordyceps sinensis can produce anti-asthmatic effect, which may play a role in allergen-induced asthma therapy.

제주도 가임연령 여교사의 톡소포자충 항체 양성률 (The Seroprevalence of Toxoplasma gondii Iefection in Teachers of Child-bearing Age in Cheju Island)

  • 양현종;홍성철;배종면
    • Journal of Preventive Medicine and Public Health
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    • 제34권4호
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    • pp.444-446
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    • 2001
  • Objectives : Toxoplasmosis is a member of the zoonosis group and may cause congenital infection . Antibody positive rates of toxoplasmosis were examined in high school students in Cheju, Korea to facilitate the study aim of examining the seroprevalence of Toxoplasma gondii in school teachers of child-bearing age in Cheju Island. Methods : The study population comprised teachers of child-bearing age in primary, middle and high schools, aged 35 years and younger, who wished to be tested for Toxoplasma gondii antibodies (IgG) by the indirect latex agglutination test (ILA) and the enzyme-linked immunosorbent assay (ELISA) method. Results : The overall antibody positive rate was 3.8% in the study subjects (n=314), a rate which showed no significant difference due to birth place, history of bringing up pets, or history of contacting a cat. Conclusion : We confirmed that the seroprevalence of Toxoplasma gondii in a population of child-bearing teachers in Cheju Island was the same as that previously reported in other parts of Korea.

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매운맛 선호도가 사람의 말초혈액에서 불리한 면역세포 활성에 미치는 영향 (Effect of Hot Taste Preference on Selected Immune Responses in Human Peripheral Immunocompetent Cells)

  • 표종옥;한인섭;김병삼;유리나
    • 한국식품영양과학회지
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    • 제26권6호
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    • pp.1194-1199
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    • 1997
  • The effect of hot taste preference on selected immune responses was investigated in human peripheral immunocompetent cells. Human lymphocytes and natural killer(NK) cells were prepared at a concentration of 2$\times$$10^{6}$ cells/ml in RPMI-1640 containing 10% fetal bovine serum. Lymphocytes proliferation was determined with the [$^{3}H$]-thymidine pulse for 18hrs after concanavalin A, phytohemagglutinin, Salmonella typhimurium mitogen, or media alone. NK cell activity was measured by cytolysis of $^51Cr$-labeled target cells K562. Serum antibodies levels such as IgM, IgG, IgA were also measured by ELISA method. There was no difference of serum IgM level among the groups, but IgG and IgA levels were greater in the group with hot taste preference than those of the group without hot taste preference. In lymphocytes of the group with hot taste preference there was a greater mitogen-induced lymphocyte proliferative responses compared to the group without hot taste preference. In addition, NK cell activity in group with hot taste preference was lower than that of the group without hot taste preference. These results suggest that the eating habit of spicy food containing hot components may affect immune status by modulating selective immunocompetent cells function.

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Ochnaflavone의 면역보조제 효과 (Effect of Ochnaflavone as An Immunoadjuvant)

  • 박민주;유기연;한용문
    • 약학회지
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    • 제56권6호
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    • pp.366-371
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    • 2012
  • In this present study, we determined whether or not there is an immunoadjuvant effect of ochnaflavone, a biflavone isolated from Lonicera japonica. As an antigenic source, the cell wall (CACW) of Candida albicans, a fungal pathogen, was used. CACW consists of 95% carbohydrate (mannan). In the experiments, BALB/c mice were immunized with emersion forms of CACW combined with or without ochnaflavone (Och) in the presence of IFA containing mineral oil or CACW alone. Then, the amounts of antisera collected from these mice groups were measured by the ELISA method. Data from these experiments showed that CACW combined with Och (CACW/Och/IFA) provoked the production of antisera app. 2.2 or 5 times more than the corresponding CACW/IFA or CACW alone (CACW/DPBS), respectively, in mice (P<0.05). We further examined the immune response type induced by Och. Analysis of the values of the IgG1/IgG2a ratios obtained from IgG isotyping revealed that Och induced Th2-immunity more dominantly than Th1. This finding was confirmed by cytokine profile. CACW/Och/IFA formulation induced IL-4 (Th2-type cytokine) more than IFN${\gamma}$ (Th1-type cytokine) as compared with CACW/IFA and CACW/DPBS formulations (P<0.05). All data combined, Och appears to have an immunoadjuvant activity that may convert Th1 immunity into Th2 immunity.

상황버섯 추출물이 항염증 및 항알레르기, 항산화, 주름제거에 미치는 실험적 연구 (Effect of Phellinus igniarius Quel Extract on the Anti-inflammatory, Anti allergy, Anti-oxidant, Anti-wrinkle)

  • 윤우식;정현아;노석선
    • 한방안이비인후피부과학회지
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    • 제23권1호
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    • pp.75-93
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    • 2010
  • Objectives : This study was carried out to investigate the effects of Phellinus igniarius Quel extract on the Anti-inflammatory, Anti allergy, Anti-oxidant and Anti-wrinkle. Methods : To investigate in vitro anti-oxidant activity assay, ethanol extracts of medicinal plants tested by DPPH method. In the next experiment, to investigate anti-inflammatory test, the RAW 264.7 macrophage cells was cultured using DMEM including the 10% FBS. To study anti-allergic effect, we blended cultured Human Mast Cells(HMC-1), and then observe TNF-$\alpha$, IL-8 by ELISA Results : Phellinus igniarius Quel extract has the effects of anti-inflammation and anti-allergy, which may be due to its inhibitory potential on the macrophage activation. Furthermore, Phellinus igniarius Quel extract has the anti-wrinkle effects through the inhibitory potential on the collagnease, elastase and gelatinase activities. Conclusions : The above results suggest that Phellinus igniarius Quel extract could be applicable for improvement of several skin functions.

Noninvasive Detection of Specific Diagnostic Biomarkers for Atopic Dermatitis

  • Chang, Jeong Hyun
    • 대한의생명과학회지
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    • 제25권1호
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    • pp.15-22
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    • 2019
  • The diagnosis of atopic dermatitis (AD) includes a test that checks allergen-mediated skin reactions and a method of measuring the total IgE and allergen-specific IgE in blood. However, these test methods are performed directly on the patient, which cause some pain or discomfort. In addition, the skin response test or IgE may result in false negative in about 20% of patients. In the present study, to identify specific biomarkers, HaCaT cells were used as a human keratinocyte that make up the skin, were treated IL-4 and IL-13 for 24 hours to induce a situation similar to keratinocytes in AD patients. In the HaCaT cells, pro-inflammatory cytokine such as IL-5, IL-6, and MCP-1 were increased by IL-4 and IL-13 and skin barrier proteins was reduced by IL-4 and L-13. This results showed that a situation similar to the stratum corneum of an actual patient is induced in HaCaT cells. And then the secretions of Kallikrein (KLK) 5 and KLK7 protease were checked by enzyme-linked immunosorbent assay (ELISA). It was specifically increased by IL-4 and IL-13. This showed that AD-related protease can be detected at the protein level using keratinocytes that can be taken in a non-invasive manner and suggested the possibility of applying it to AD diagnosis.

Daily Amperometric Monitoring of Immunoglobulin E in a Mouse Whole Blood: Model of Ovalbumin Induced Asthma

  • Lee, Ju Kyung;Yoon, Sung-hoon;Kim, Sang Hee
    • 전기화학회지
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    • 제25권1호
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    • pp.13-21
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    • 2022
  • There is an increasing interest in monitoring of specific biomarker for determining progression of a disease or efficacy of a treatment. Conventional method for quantification of specific biomarkers as enzyme linked immunosorbent assay (ELISA) has high material costs, long incubation periods, requires large volume of samples and involves special instruments, which necessitates clinical samples to be sent to a lab. This paper reports on the development of an electrochemical biosensor to measure total immunoglobulin E (IgE), a marker of asthma disease that varies with age, gender, and disease in concentrations from 0.3-1000 ng/mL with consuming 20 µL volume of whole blood sample. The sensor provides rapid, accurate, easy, point-of-care measurement of IgE, also, sequential monitoring of total IgE with ovalbumin (OVA) induced mice is another application of sensor. Taken together, these results provide an alternative way for detection of biomarkers in whole blood with low volumes and long-term ex-vivo assessments for understanding the progression of a disease.