• Title/Summary/Keyword: ELISA kit

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Development of COVID-19 Neutralizing Antibody (NAb) Detection Kits Using the S1 RBD Protein of SARS-CoV-2 (코로나 바이러스 감염증-19의 재조합 S1 RBD 단백질을 이용한 COVID-19 바이러스의 중화항체 검사 키트의 개발)

  • Choi, Dong Ok;Lee, Kang Moon
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.3
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    • pp.257-265
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    • 2021
  • The COVID-19 virus is a β-genus virus that causes infection by mediating the angiotensin convertible enzyme 2 (ACE2) receptor, which is distributed in large numbers in the human respiratory tract. The disease requires effective post-management of antibody production by complete healers and vaccinators because there is no perfect remedy for the virus infection. This study aimed to develop recombinant proteins specifically responsive to neutralizing antibodies in clinical specimens and use them to develop a rapid diagnostic kit to diagnose neutralizing antibodies quickly and conveniently against the COVID-19 virus and confirm the possibility of commercialization through a performance evaluation. Rapid diagnostic kits using COVID-19 S1 RBD recombinant proteins can be applied to rapid diagnostic kits, with positive percentage agreement (PPA) and negative percentage agreement (NPA) of 100% and 98.3%, respectively, compared to the U.S. FDA-approved ELISA kits. If the performance of the rapid diagnostic kit is improved and neutralizing antibodies can be analyzed quantitatively using quantitative analysis equipment, it can be used as important data to predict immunity to the COVID-19 virus and determine additional vaccinations.

The Effects of Dictamni Radicis Cortex on the iNOS Expression and Proinflammatory Cytokines Production (백선피의 iNOS발현과 염증성사이토카인의 생성에 미치는 영향)

  • Park, Jeong-Suk;Shin, Tae-Yong;Kim, Dae-Keun;Lee, Jae-Hyeok
    • Korean Journal of Pharmacognosy
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    • v.42 no.4
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    • pp.348-353
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    • 2011
  • The aim of the present study is to investigate the cytokine production inhibitory effect of a Dictamni Radicis Cortex (DRC). DRC has been commonly used as important medicinal herb in China and it used to control eczema, atopic dermatitis, fever and inflammatory diseases. Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the DRC on proinflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of DRC, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the DRC reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the DRC may have an anti-inflammatory property through suppressing inflammatory mediator productions.

Enhancing effect of Canavalia gladiata DC semen on the hematopoietic expansion and function of stem cells (도두(刀豆)의 조혈증진 작용에 관한 연구)

  • Kim, Seung-Hyung;Kim, Kun-Hoae;Chi, Gyeong-Yup;Cho, In-Sik;Kim, Han-Young;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.27 no.4
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    • pp.9-16
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    • 2012
  • Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

Osteogenic differentiation of bone marrow derived stem cells in gelatin-hydroxyapatite nanocomposite

  • Jeon, Hyun-Jun;Hwang, Young-Sup;Kim, Uk-Kyu;Hwang, Dae-Seok;Lee, Kwang-Ho;Chang, Myung-Cheol
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.1
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    • pp.7-12
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    • 2009
  • Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores ($20-40{\mu}m$). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.

Pentoxifylline Induces Lipolysis and Apoptosis of Human Preadipocytes, Keratinocytes and Fibroblasts In Vitro

  • Lee, Il-Kyu;Choi, Yun-Jung;Shim, In-Sop;Kim, Kyung-Soo;Choi, Chang-Jin
    • Biomolecules & Therapeutics
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    • v.18 no.1
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    • pp.56-64
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    • 2010
  • Pentoxifylline (PTX) has been used for the local reduction of fat tissue in the clinical setting. However, its safety and efficacy have not been proven. The aim of this study was to evaluate the effects of PTX on cell lines established from fat tissue. Newly cultured human preadipocytes and adipocytes from subcutaneous abdominal fat in addition to purchased human lung fibroblasts and keratinocytes were treated with PTX at different concentrations. Cell viability was determined using the Cell counting kit (CCK)-8 assay and lipolysis was evaluated using an Elisa kit. DNA fragmentation, Western blot analysis, Hoechst and Propidium Iodide (PI) staining and fluorescence activated cell scanning analysis were performed to confirm apoptosis. The viability of adipocytes, preadipocytes, keratinocytes and fibroblasts was markedly decreased at concentrations of PTX above 20 mM. Apoptosis was induced at concentrations of PTX over 40 mM in all cell lines. Lipolysis was increased by 60% at concentrations of PTX of 20 mM compared to the control. In conclusion, the results of this study showed that 20 mM of PTX induced lipolysis. At concentrations over 20 mM, PTX reduced the viability of all cells studied including: adipocytes, preadipocytes, fibroblasts and keratinocytes, in a non-specific manner.

A Study of the Inhibitory Effect of Acer tegmentosum Max. on Fibrogenesis in Hepatic Stellate Cell Line T6 (산청목(山靑木)이 간섬유화 진행 억제에 미치는 효과에 대한 연구)

  • Lee, Seung-Bo;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.346-355
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    • 2010
  • Objectives : This study was performed in order to investigate the anti-fibrogenic effect of Acer tegmentosum Maxim. on r at hepatic stellate cell line T6. Materials and Methods : Hepatic stellate Cells (T6) were treated with various concentrations of distilled water Acer teg mentosum Maxim. extract for 24, 48, 72 hours. After the treatment, cell viability, proliferation, procollagen levels, mRNA of AS MA, MMP-2, collagen type 1a2 and IL-6 production were measured using MTT assay, BrdU assay, RT-PCR, procollagen typ e 1 C-peptide EIA kit and murine IL-6 ELISA development kit. Results : Cell viability of HSC-T6 decreased significantly in both 24 hours and 48 hours groups in a dose-dependant man ner. Proliferation of HSC also decreased in the same way. In the RT-PCR, mRNA expression of collagen type 1a2 and ASMA decreased in the groups which were treated with Acer tegmentosum Maxim. for 24 hours. The production of procollagen tended to decrease in a dose-dependant manner in the 24 hours treated group. IL-6 production increased under Acer tegmentosum trea tment in a dose-dependant manner in both 24 and 48 hours groups. Conclusion : These results show the possibility that Acer tegmentosum Maxim. can be an effective remedy for liver fibrosi s and liver cirrhosis.

Productivity, Isolation and Purification of Egg Yolk Antibody(IgY) againt Food Poisoning Bacteria (Salmonella typhimurium) (식중독균 항원(Salmonella typhimurium)에 의한 계란항체(IgY) 생산성과 분리 정제)

  • 한준표;백반석;배만종
    • Journal of the East Asian Society of Dietary Life
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    • v.9 no.2
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    • pp.200-206
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    • 1999
  • This study was carried out to get a industrial information about a possibility of IgY antibody production, antimicrobial activity and Properties of IgY antibody in egg yolk. After the initial immunization the anti-Salmonella typhimurium IgY antibody level gradually were decreased from firth week to tenth week. On the other hand, the antibody level in the serum were increased from the first week, reaching its peak in the sixth week. Molecular weights of IgY were estimated approximately 72-75KD in a heavy chain and 30-40KD in a light chain by electrophoresis.

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Anti-adipogenesis Effects of 3 Herbal Formula on Blood Stasis (어혈처방 3종이 지방전구세포 분화에 미치는 영향)

  • Lee, Hoyoung;Shim, Eun Hyoung;Lee, Myeong Soo;Lee, Ju Ah
    • The Korea Journal of Herbology
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    • v.31 no.6
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    • pp.29-35
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    • 2016
  • Objectives : Blood stasis (BS) is related to be caused by blood circulation and stagnation which are cancer, atherosclerosis and hyperlipidemia in traditional medicine. We extracted 3 kinds of BS formula; Seogakjihwang-tang (SGT), Tonggyuhawlhyul-tang (THT), Hyulbuchukeo-tang (HCT). This study was conducted to investigate whether the 3 kinds of herbal formula extracts have inhibitory efficacy association with anti-adipogenesis. Methods : To investigate the anti-adipogenesis, we used the mouse fibroblast cell line, 3T3-L1 which differentiated into adipocytes in response to insulin, IBMX and dexamethasone (MDI). Cytotoxicity of herbal formula extracts were examined by CCK-8 kit. Intracellular lipid droplets were detected by Oil-Red-O staining. Triglyceride (TG) and leptin were measure using elisa kit. Results : The yield of water extracts was 14.62% (SGT), 21.27% (THT), 20.02% (HCT). Lipid accumulation was reduced significantly by 3 kinds of herbal formula compared to control. Especially, THT and HCT decreased lipid droplet, respectively at all concentration. The TG and leptin were also inhibited by 3 kinds of herbal formula. The IC50 of TG were $280.51{\mu}g/m{\ell}$ (SGT), $52.62{\mu}g/m{\ell}$ (THT), $313.99{\mu}g/m{\ell}$ (HCT). The IC50 of leptin were $348.76{\mu}g/m{\ell}$ (SGT), $164.02{\mu}g/m{\ell}$ (THT), $257.00{\mu}g/m{\ell}$ (HCT). THT was better than other herbal formula on anti-adipogenesis. Conclusion : kinds of herbal formula inhibited adipogenesis in MDI-induced 3T3-L1 adipocytes, as indicated by the significant reduction in TG and leptin concentration without cytotoxicity. Therefore, 3 kinds of herbal formula for BS might act as a therapeutic agent for preventing lipid diseases, such as obesity and atherosclerosis.

Anti-inflammatory Activity of Herbal Extracts through Inhibition of TNF-α, IL-6 and IL-8 (TNF-α, IL-6, IL-8 저해를 통한 천연약용식물 추출물의 항염증 특성)

  • Kim, Yong-Soo;Gwon, Hui-Jeong;Park, Jong-Seok;Lim, Youn-Mook;Nho, Young-Chang
    • Journal of Radiation Industry
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    • v.5 no.3
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    • pp.273-277
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    • 2011
  • This study evaluated the cytotoxicity and anti-inflammatory activities of natural herbal extracts including Houttuynia cordata and Eucommia ulmoides against human mast cell (HMC-1). Houttuynia cordata (HC) and Eucommia ulmoides(EU) were extracted with distilled water (at $75^{\circ}C$) and then freeze-dried for 5 days. Finally, the mixture of HC and EU were sterilized by ${\gamma}$-rays irradiation. Cytotoxicity of the mixture against HMC-1 cell was measured using cell counting kit-8 (CCK-8) assay. In addition, inflammatory mediator cytokines such as TNF-${\alpha}$, IL-6 and IL-8 were evaluated by ELISA kit on the HMC-1 cells with calcimycin A23187 and phorbol 12-myristate 13-acetate (PMA). The results showed that mixture of HC and EU had no cytoxicity and reduced TNF-${\alpha}$, IL-6, IL-8 response on HMC-1 cells.

In Vitro Anti-inflammatory Activities of Herbal Extracts with Eucommia ulimoides and Ulmus davidiana (두충과 느릅 혼합추출물에 의한 염증 억제 효과)

  • Gwon, Hui-Jeong;Shin, Young Min;Jung, Sung-In;Park, Jong-Seok;Nho, Young-Chang;Lim, Youn-Mook
    • Journal of Radiation Industry
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    • v.6 no.1
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    • pp.11-15
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    • 2012
  • This study confirmed the cytotoxicity and anti-inflammatory activities of natural herbal extracts (HE) including Eucommia ulimoides and Ulmus davidiana against human mast cell (HMC-1). HE was extracted with distilled water (at $75^{\circ}C$) and then freeze-dried for 5 days. Finally, the HE was sterilized by gamma radiation with $^{60}Co$ ${\gamma}$ source at room temperature. Cytotoxicity of the HE against HMC-1 cell was measured using cell counting kit-8 (CCK-8) assay. In addition, inflammatory cytokines such as $TNF-{\alpha}$, IL-6 and IL-8 were evaluated by ELISA kit on the HMC-1 cells with calcimycin A23187 and phorbol 12-myristate 13-acetate (PMA). The results showed that HE had no toxicity and reduced $TNF-{\alpha}$, IL-6, IL-8 response on HMC-1 cells.