• Title/Summary/Keyword: neutralizing activity

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Production of the Monoclonal Antibodies to the Escherichia coli Heat-Stable Enterotoxin (대장균의 내열성장독소 측정법개발을 위한 단세포군항체의 생산)

  • Chang, Woo-Hyun;Lee, Woo-Kon;Kim, Suck-Yong;Park, Jung-Bum
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.4
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    • pp.377-392
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    • 1987
  • Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.

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Evaluation of Neutralizing Capacities of Antacid Products (제산제의 중화 능력의 평가 연구)

  • 박경호;차수만;최진석;김낙두
    • YAKHAK HOEJI
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    • v.27 no.2
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    • pp.139-148
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    • 1983
  • The neutralizing capacities of the antacids, which are frequently used in Korean market, were evaluated in vitro by the methods of Resset and Rice, Fordtran and Collyns, and modified Beekman, respectively. The antacids used in the study are two kinds, the one is preparations from Seoul National University Hospital and the other is products from pharmaceutical companies, and their components are aluminum phosphate, aluminum hydroxide, magnesium aluminum hydroxide, magnesium hydroxide, basic aluminum sucrose sulfate and $2MgO{\times}Al_{2}O_{3}{\times}SiO_{3}$, etc. The hospital preparations, DMC and MAC powders, showed most powerful and sustained neutralizing capacities, i.e. they maintained the pH range from 5 to 8 for 60min, Whereas pharmaceutical products, aluminum hydroxide gel containing magnesium hydroxide and magnesium aluminum hydroxide gel exhibited a moderate capacities, i.e pH ranged from 3 to 6, and aluminum phosphate, $2MgO{\times}Al_{2}O_{3}{\times}SiO_{2}$ and basic aluminum sucrose sulfate displayed a weak activity, pH ranged from 2 to 3. When the therapeutic doses of aluminum hydroxide gel containing magnesium hydroxide and magnesium aluminum hydroxide gel were divided into 2 doses and each dose was used at the interval of 30min., the divided doses kept more prolonged higher pH than the single therapeutic dose. Milliequivalents of neutralizing capacities of each antacid were measured by the method of Fordtran and Collyns. The milliequivalents per 1ml of aluminum hydroxide gel, aluminum hydroxide gel containing magnesium hydroxide, magnesium aluminum hydroxide gel and aluminum phosphate were 2.87, 2.86, 2.57, and 0.67, respectively. The milliequivalents per 100mg of preparations, i.e. MAC powder, dried aluminum hydroxidgel, DMC powder, 2MgO, $Al_{2}O_{3}$. $SiO_{2}$, magnesium aluminum hydroxide and basic aluminum sucrose sulfate were 1.91, 1.68 1.63, 1.45, 1.44, and 0.44, respectively.

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Evaluation for Protective Effect of Rutin, a Natural Flavonoid, against HCl/Ethanol-Induced Gastric Lesions

  • Jeong, Choon-Sik
    • Biomolecules & Therapeutics
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    • v.17 no.2
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    • pp.199-204
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    • 2009
  • In this study we investigated the protective effects of rutin, a natural plant flavonoid, on HCl/ethanol-induced gastric lesions in rats. Rutin showed the antioxidant activities, the acid-neutralizing capacities, and the inhibitory effects on the growth of Helicobacter pylori (H. pylori.), which are equivalent to control compounds. In addition, rutin significantly inhibited HCl/ethanol-induced gastric lesions. Antigastritic action of rutin may be associated with the antioxidant activities, acid-neutralizing capacities, anti-H. pylori action, and the stimulation of mucus secretion. From these results, we could suggest that rutin may be useful for the treatment and/or protection of gastritis.

Production and characterization of monoclonal antibodies against porcine transmissible gastroenteritis virus (돼지 전염성 위장염 바이러스에 대한 단크론항체 생산 및 특성)

  • Jang, Young-en;Cho, Sun-hee;Kim, Byung-han;Ahn, Jae-moon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.336-344
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    • 1998
  • Eight monoclonal antibodies(MAbs) against the transmissible gastroenteritis virus (TGEV) were produced and characterized. Four of the MAbs were produced against a reference TGEV, Purdue strain(P115) and the others were produced against the Korean vaccine virus, Pyungtaek strain. Only one MAb(5C8) produced against P115 had neutralizing activity and was found to be E2 protein-specific. The other seven MAbs(4E2, 4G8, 5H6, 1F8, 2C6, 5H5, and 3A6) had specificity of nucleocapsid protein and no neutralizing activity. All MAbs reacted with different strains of TGEV, but none of the MAbs was reactive with porcine enteropathogenic viruses such as rotavirus, epidemic diarrhea virus and enterovirus by fluorescence antibody(FA) test.

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THE EFFECT OF GROWTH FACTORS IN PLATELET-RICH PLASMA ON THE ACTIVITY OF OSTEOBLAST CELL LINE (혈소판농축혈장 내의 성장요소가 조골세포주의 활성도에 미치는 영향)

  • Jung Tae-Wook;Kim Yung-Soo;Kim Chang-Whe;Jang Kyung-Soo;Lim Young-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.2
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    • pp.175-191
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    • 2004
  • Statement of problem: Platelet-rich plasma(PRP) is well known to be very effective method to stimulate and accelerate the healing of bone and soft tissue. However, there are few reports which deal with the mechanisms of the PRP on the activation of the osteoblasts. Purpose: This study was aimed to investigate the effect of growth factors in PRP on the activity of osteoblasts. Material and method: To evaluate the effect on human, human osteoblast cell line was cultured. PRP was extracted from the blood of a healthy volunteer. Using the recombinant growth factors of PDGF, $TGFT-\beta$, IGF-1, bFGF which are mainly found at bone matrix and their neutralizing antibody, the effect of PRP on the attachment and proliferation of osteoblasts was evaluated. To evaluate the autocrine and paracrine effects, conditioned media(CM) of PRP was made and compared with PRP. By the western blot analysis, the expression of growth factors in PRP, CM was examined. Cell morphology was compared by the light microscope. Results : 1) The effects of CM on osteoblast were similar to the effects of PRP. 2) PRP, CM, recombinant $TGF-\beta$, bFGF, IGF-1 showed significantly higher cellular attachment than control(p<0.05) in the cell attachment assay. In the cell proliferation assay, PRP, CM, recombinant $TGF-\beta$, IGF-1, bFGF, PDGF increased significantly cell proliferation(p<0.01). Among the recombinant growth factors, IGF-1 showed the highest cellular attachment and proliferation. 3) In the western blot assay, bFGF, IGF-1, PDGF weve equally expressed in PRP and CM. 4) The attachment of osteoblast cell decreased significantly after the addition of neutralizing antibody against $TGF-\beta$, IGF-1(p<0.05). In the cell proliferation assay, the addition of neutralizing antibody against $TGF-\beta$, bFGF, PDGF, IGF-1 decreased significantly the cellular proliferation(p<0.05). The amount of decreasing in the cell attachment and proliferation is the highest in at-lGF-1. 5) The cells in control group were flattened and elongated with a few cellular processes in the a light microscope. But, the cells appeared as spherical, plump cells with well developed cellular processes in experimental groups. The cells in PRP and CM had more prominent developed features than recombinant growth factor groups. Conclusions : These findings imply that PRP maximize the cellular activity in early healing period using the synergistic effect, autocrine, paracrine effects of growth factors and increase the rate and degree of bone formation.

Immunogenicity of Recombinant Human Erythropoietin: Clinical Cases, Causes and Assays

  • Heo, Tae-Hwe;Kim, Young-Kwon;Yang, Seung-Ju;Cho, Hyun-Jeong;Kim, Sung-Jo
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.161-166
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    • 2009
  • Human erythropoietin(EPO) is a glycoprotein that enhances red blood cell production by stimulating proliferation and differentiation of erythroid progenitor cells in the bone marrow. Patients with chronic kidney disease(CKD) suffer from anemia caused by reduced production of EPO in the kidney. Recombinant human EPO protein has been used successfully for the treatment of anemia associated with CKD. Recently, attention has been paid to the development of side effect of EPO, pure red cell aplasia(PRCA), in some patients with CKD. PRCA is a rare disorder of erythropoiesis that leads to a severe anemia due to an almost complete cessation of red blood cell production. EPO-related PRCA is caused by the production of EPO-neutralizing antibodies(Abs) that eliminate the biological activity of EPO as well as endogenous EPO in patients undergoing therapy. Since 1988, almost 200 cases worldwide have been reported with Ab-positive PRCA after receiving EPO therapeutics. The underlying mechanisms of the breaking of immune tolerance to self-EPO have been investigated. Modification of formulation, organic compounds of container closures, and route of administration has been suggested for the possible mechanism of increased immunogenicity of EPO. A number of assays have been used to detect Abs specific to EPO. These assays are generally grouped into two major categories: binding Ab assays and neutralizing Ab assays(bioassays). There are several types of binding Ab assays, including radioimmunoprecipitation assay, enzyme-linked immunosorbent assay, and the BIAcore biosensor assay. In vitro cell-based bioassays have been utilized for the detection of neutralizing Abs. Finally, the recent experience with anti-EPO Abs may have considerable implications for the future development and approval of EPO preparations. Also, considering that millions of patients are being treated with EPO, clinicians need to be aware of signs and consequences of this rare but severe clinical case.

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A Review on Venom Enzymes Neutralizing Ability of Secondary Metabolites from Medicinal Plants

  • Singh, Pushpendra;Yasir, Mohammad;Hazarika, Risha;Sugunan, Sunisha;Shrivastava, Rahul
    • Journal of Pharmacopuncture
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    • v.20 no.3
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    • pp.173-178
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    • 2017
  • Objectives: Medicinal plants are vital sources of bioactive compounds that are useful for the treatment of patients with snake bites or are indirectly applicable for boosting the effects of conventional serum therapy. These plants are being used traditionally by local healers and tribes for the treatment of patients with snake bites and therefore can be used as an alternative against snake envenomation. Scientifically, using the secondary metabolites of plants to neutralize venom enzymes has an extra benefit of being based on traditional knowledge; also, the use of such metabolites for the treatment of patients with snake bites is cheaper and the treatment can be started sooner. Methods: All the available information on various secondary metabolites exhibiting venom neutralizing ability were collected via electronic search (using Google books, Pubmed, SciFinder, Scirus, Google Scholar, and Web of Science) and articles of peer-reviewed journals. Results:Recent interest in different plant has focused on isolating and identifying of different phytoconstituents that exhibit Phospholipase A2 activity and other venom enzyme neutralizing ability. In this support convincing evidence in experimental animal models are available. Conclusion: Secondary metabolites are naturally present, have no side effect, are stable for a long time, can be easily stored, and can neutralize a wide range of snake enzymes, such as phospholipase A2, hyaluronidase, protease, L-amino acid oxidase, 5'nucleotidase, etc. The current review presents a compilation of important plant secondary metabolites that are effective against snake venom due to enzyme neutralization.

Immune responses to synthetic peptides of RSV F protein

  • Kim, Jung-Kwon;Lee, Hoan-Jong;Kim, Hong-Jin
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.310.1-310.1
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    • 2002
  • The fusion (F) protein of respiratory syncytial virus(RSV) is an important antigen in including cross-protective immunity with neutralizing activity. Two peptides homologous to amino acid 205-225(F/205-225) and 255-278 (F/255-278) of the F glycoprotein of RSV containing B cell and T cell epitope were synthesized and then conjugated with KLH. (omitted)

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Effects of Ligustrum Lucidum on the Phagocytic Activity of Macrophages (여정자(如貞子)가 대식세포(大食細胞) 탐식능(貪食能)에 미치는 영향(影響))

  • Lee, Kwang-Souk;Song, Bong-Keun;Kim, Hyeong-Kyun;Lee, Eon-Jeong
    • The Journal of Korean Medicine
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    • v.17 no.2 s.32
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    • pp.227-236
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    • 1996
  • The effect of Ligustrum Lucidum(LL) on the production of nitric oxide (NO) and superoxide by murine peritoneal macrophages were investigated. Stimulation of the cells with LL in the presence or absence of interferon-r(IFN-r) resulted in the increased accumulation of nitrite in the medium. To further examine the mechanism of LL induced. NO Synthesis, we evaluated the secretion of tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ by LL in murine macrophages. Treatment of LL increased the secretion of bioactive $TNF-{\alpha}$ in cultured medium. In addition, LL induced NO production was decreased by the treatment of anti-murine $TNF-{\alpha}$. neutralizing antibodies, indicating that LL induced superoxide production was decreased by the treatment of anti-murine $TNF-{\alpha}$ neutralizing antibodies. These data suggested that LL induced superoxide production was related to $TNF-{\alpha}$ secretion. In conclusion, our results indicates that LL may enhance innate immune response and be applied as a immunoregulating drug improving phagocytosis.

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