• 제목/요약/키워드: antibody formation

검색결과 187건 처리시간 0.031초

백서의 골 결손부 치유시 Transforming Growth Factor-β의 발현에 대한 면역 조직 화학 연구 (IMMUNOHISTOCHEMICAL STUDY OF TGF-β ON THE HEALING PROCESS IN THE RAT FEMUR BONE DEFECT)

  • 방승준;김경욱;이재훈
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • 제20권2호
    • /
    • pp.173-183
    • /
    • 1998
  • $TGF-{\beta}$ is one of growth factors that may be involved in the formation of bone and cartilage. Multiple studies demonstrate that $TGF-{\beta}$ is involved in regulating cell proliferation, differentiation and matrix synthesis, events observed in frature healing. The apperance of $TGF-{\beta}$ in the fracture during healing was evaluated by immunohistologic localization of $TGF-{\beta}$ using antibody. Twenty Sprauge-Dawley strain white male rats, each weiging about 150grams were used and divided two groups. The one group, the $2{\times}2mm$ bony defect was formed in the right femur. The other group, $4{\times}2mm$ bony defect was formed in right femur. Both group were sacrificed at 3day, 1, 2, 3, 4 week and femur were harvested, paraffin sections were stained with H & E, MT stain, immunihistochemical staining with $TGF-{\beta}$ antibody and observed under light microscope. The result were as follows: 1. New bone formation and cartilagenous tissue was seen at 3day. And in the $2{\times}2mm$ bony defect group, $TGF-{\beta}$ stained the cell surounding new bone. 2. The osteoclast and trabecular was seen at 1week. $TGF-{\beta}$ stained the osteoblast and in the $2{\times}2mm$ bony defect group was stained more than $4{\times}2mm$ bony defect group. 3. The lamella bone and trabecule was seen from 3, 4week, and $TGF-{\beta}$ stained almost negative. From the above finding, we could concluded that $TGF-{\beta}$ stained the osteoblast at early stage and 1week, the peak stain was seen from 1week, and then decreased, almost negative stain was seen at 3, 4week.

  • PDF

Formation of Cross-Linked Products of The Reaction Center D1 Protein in Photosystem II under Light Stress

  • Uchida, Suguru;Kato, Yoji;Yamamoto, Yasusi
    • Journal of Photoscience
    • /
    • 제9권2호
    • /
    • pp.382-384
    • /
    • 2002
  • When illuminated with strong visible light, the reaction center Dl protein of photo system II is photodamage and degraded. Reactive oxygen species and endogenous cationic radicals generated by photochemical reactions are the cause of the damage to the Dl protein. Recently we found that the photodamaged Dl protein cross-links with the surrounding polypeptides such as D2 and CP43 in photosystem II. As the cross-linking reaction is dependent on the presence of oxygen, reactive oxygen species are suggested to be involved. Among the reactive oxygen species examined, ? OH was most effective in the formation of the cross-linked products. These results indicate that the cross-linking is mostly due to ? OH generated at photosystem II. The cross-linking site of the Dl protein is not known. As several tyrosine residues exist at the D­E loop of the Dl protein, there is a possibility that di-Tyr is formed between the D­E loop of the Dl protein and surrounding polypeptides during the strong illumination. Therefore, we examined the formation of di-Tyr using the monoclonal antibody against di-Tyr under excess illumination of the photosystem II membranes. The results obtained here suggest that no di-Tyr is formed during the excess illumination of photosystem II.

  • PDF

Development of Surface Plasmon Resonance Immunosensor through Metal Ion Affinity and Mixed Self-Assembled Monolayer

  • Lee, Si-Ra;Sim, Sang-Jun;Park, Chul-Hwan;Gu, Man-Bock;Hwang, Un-Yeon;Yi, Jong-Heop;Oh, Byung-Keun;Lee, Jin-Won
    • Journal of Microbiology and Biotechnology
    • /
    • 제18권10호
    • /
    • pp.1695-1700
    • /
    • 2008
  • An immunosensor based on surface plasmon resonance (SPR) with enhanced performance was developed through a mixed self-assembled monolayer. A mixture of 16-mercaptohexadecanic acid (16-MHA) and 1-undecanethiol with various molar ratios was self-assembled on gold (Au) surface and the carboxylic acid groups of 16-MHA were then coordinated to Zn ions by exposing the substrate to an ethanolic solution of $Zn(NO_3)_2\cdot 6H_2O$. The antibody was immobilized on the SPR surface by exposing the functionalized substrate to the desired solution of antibody in phosphate-buffered saline (PBS) molecules. The film formation in series was confirmed by SPR and atomic force microscopy (AFM). The functionalized surface was applied to develop an SPR immunosensor for detecting human serum albumin (HSA) and the estimated detection limit (DL) was 4.27 nM. The limit value concentration can be well measured between ill and healthy conditions.

Enhanced Tumor-targeted Gene Delivery by Immunolipoplexes Conjugated with the Humanized Anti-TAG-72 Fab' Fragments

  • Kim, Keun-Sik;Park, Yong-Serk;Hong, Hyo-Jeong;Kim, Kwang-Pyo;Lee, Kwang-Hyun;Kim, Dong-Eun
    • Bulletin of the Korean Chemical Society
    • /
    • 제33권2호
    • /
    • pp.651-656
    • /
    • 2012
  • Cationic immunoliposomes were prepared by conjugation of Fab' fragments of the recombinant humanized monoclonal antibody (HuCC49) against tumor-associated glycoprotein (TAG)-72 to sterically unilamella liposomes. The cationic immunoliposomes are composed of cationic lipid (O,O'-dimyristyl-N-lysyl aspartate, DMKD), cholesterol, and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethyleneglycol)$_{2000}$] (DPPE-PEG-maleimide) with a molar ratio of 0.5:0.47:0.03. Plasmid DNA was effectively condensed by addition of transferrin (Tf) during the formation of anti-TAG-72 PEG-immunolipoplexes (PILPs). These anti-TAG-72 PILPs were able to adhere to the surface of TAG-72-overexepressing LS174T human colon cancer cells more effectively than conventional liposomes, thereby facilitating gene delivery in vitro. Furthermore, intravenous administration of the anti-TAG-72 PILPs into the tumor-carrying mice exhibited efficient localization of the reporter gene in the tumor tissues.

Propagation of lymphocystis disease virus (LCDV) in the FIN cell line originated from olive flounder Paralichthys olivaceus fin

  • Hossain, Mosharrof;Kim, Wi-Sik;Jung, Sung-Ju;Oh, Myung-Joo
    • 한국어병학회지
    • /
    • 제24권2호
    • /
    • pp.47-51
    • /
    • 2011
  • The present study demonstrated lymphocystis disease virus (LCDV) propagation through cytopathic effects (CPE) formation and LCDV detection in olive flounder fin (FFN) cells by polymerase chain reaction (PCR) and fluorescent antibody technique (FAT) methods. Tissue filtrates from the cluster cells produced CPE in FFN cells, which initially cells became enlarged and gradually underwent fusion en masse. Infectivity of culture grown LCDV using the FFN cells reached $10^{2.3}$ $TCID_{50}$/ml at 4 days post infection and the highest titer was measured $10^{6.5}$ $TCID_{50}$/ml at 12 days. The viral DNA was detected in the cell culture supernatants showing CPE and the CPE cells by PCR. Antigen specific strong fluorescence reacting with monoclonal antibody against the virus revealed the presence of viral antigen in the cytoplasm of infected FFN cells. These results suggest that the FFN cell line originated from the olive flounder has a susceptibility of the LCDV.

귀비온담탕(歸脾溫膽湯)이 흰쥐의 항(抗)스트레스와 면역반응(免疫反應)에 미치는 영향(影響) (The Effect of Guibiondamtang on Immune Response and in Concertration of Catecholamine in Immobilization Stressed Rates)

  • 조진영;황의완
    • 동의신경정신과학회지
    • /
    • 제6권1호
    • /
    • pp.1-17
    • /
    • 1995
  • In order to investigate the Anti-stress effect of Guibiondamtang in the immobilization stressed rats, the level of serum catecholamine, the change of body weight, the humoral and cellular immune response were studied. The results were as follows; 1. The decrese of the body weight was significantly inhibited in test group for Guibiondamtang, comparting to the control group. 2. The increase of the level of serum norepinephrine was significantly inhibited in test group, comparing to the control group. 3. The increase of the level of serum epinephrine was significantly inhibited in test group, comparing to the control group. 4. In the hemagglutinaton titer, the control group was decreased on the serum antibody titer but test group was inhibitory effect on the decrease of sereum antibody titer. 5. In the plaque formation test, the control and test group were not shown significant differences. 6. In the foot pad swelling respopnse, the control group was decreased on DTH response but test group was increased comparing to the normal group. 7. There was no change on the distribution of lymphocyte subset(CD4, CD8), grnulocyte and macrophage analyzed by flow cytometry.

  • PDF

Detection of Escherichia coli O157:H7 Using Immunosensor Based on Surface Plasmon Resonance

  • Oh, Byung-Keun;Kim, Young-Kee;Bae, Young-Min;Lee, Won-Hong;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
    • /
    • 제12권5호
    • /
    • pp.780-786
    • /
    • 2002
  • An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

유전자 재조합 Human galectin-3의 발현과 성상 (Expression and characterization of the recombinant human galectin-3)

  • 김병규;우희종
    • 대한수의학회지
    • /
    • 제37권3호
    • /
    • pp.547-554
    • /
    • 1997
  • Galectin-3 is known as an animal ${\beta}$-galactoside-binding lectin charicterized with S-type carbohydrate recognition domain. It plays a role in growth, adherence and movement of cells. It is, also, related to the cell transformation and metastasis of tumor cells. In this study, we have expressed and purified recombinant human galectin-3 (rHgalectin-3) using E coli system and asialofetuin affinity chromatography for the future development of monoclonal antibody to Hgalectin-3, which is suggested as the tumor marker for the gastric and thyroid gland cancers. Expressed protein was confirmed as the Hgalectin-3 by immunoblot with cross-reactive murine monoclonal antibody. Lectin activity and specificity of purified protein were, also, confirmed by the competitive inhibition with galectin-3 specific carbohydrate, lactose. Like physiological galectin-3, lectin activity of the molecule was not changed in nonreduced condition. Dimer formation, furthermore, was observed at high concentration of the protein even in the reduced condition, which is well known in physiological galectin-3. These results showed purified rHgalectin-3 has the same activity and molecular nature compared to the physiological galectin-3.

  • PDF

Purification and Characterization of Phytoferritin

  • Oh, Suk-Heung;Cho, Sung-Woo;Kwon, Tae-Ho;Yang, Moon-Sik
    • BMB Reports
    • /
    • 제29권6호
    • /
    • pp.540-544
    • /
    • 1996
  • Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.

  • PDF

FRC에서 Lymphotoxin β receptor의 자극은 MLCK와 ROCK의 이중 신호전달 경로를 통해 stress fiber 변화에 관여 (Signals of MLCK and ROCK Pathways Triggered via Lymphotoxin β Receptor are Involved in Stress Fiber Change of Fibroblastic Reticular Cells)

  • 김대식;이종환
    • 생명과학회지
    • /
    • 제29권2호
    • /
    • pp.256-264
    • /
    • 2019
  • Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$)는 TNF 계열로 림프조직의 미세구조와 기관형성에 중요한 역할을 한다. MLCK와 ROCK는 세포의 stress fiber 형성조절에 관여하는 주요 신호전달자이다. Fibroblastic reticular cell (FRC)에서 $LT{\beta}R$ 자극을 통한 이런 신호전달자들의 관련성을 알아보기 위해 ML-7 (MLCK 저해제)이 사용되었다. ML7 처리된 FRC에서 SF가 완전히 파괴되었고 anti-$LT{\beta}R$ antibody 처리 세포와 유사하게 ML7 처리 FRC에서 응축된 세포형태를 관찰 할 수 있었다. Y27632로 ROCK를 저해 했을 때 FRC의 액틴 세포골격과 세포형태 변화가 유도 되었다. FRC에서 p-MLC가 액틴과 함께 SF 구성성분을 이루었다. FRC세포 추출물로 Rho-guanosine diphosphate (GDP)/guanosine triphosphate (GTP) 교환활성을 확인했다. Agonistic anti-$LT{\beta}R$ antibody로 $LT{\beta}R$을 자극 했을 때 Rho-GDP/GTP 교환활성이 크게 감소했다. MLCK 저해처럼 $LT{\beta}R$ 자극은 MLC의 인산화를 감소시켰다. Agonistic anti-$LT{\beta}R$ antibody-treated FRC에서 세포골격 구성요소인 세포막과 세포골격 링커 역할을 하는 p-ezrin의 인산화는 감소 되었고 b- actin, 그리고 tubulin 발현도 줄었다. 이런 결과는 FRC의 $LT{\beta}R$ 신호전달을 통한 SF 조절에는 MLCK와 ROCK가 관여하고 있다는 것을 알 수 있었다.