• Title/Summary/Keyword: Antibody Engineering

Search Result 351, Processing Time 0.022 seconds

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

  • Kim, Dae Sik;Lee, Jong-Hwan
    • Journal of Life Science
    • /
    • v.29 no.2
    • /
    • pp.256-264
    • /
    • 2019
  • Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. In contrast, MLCK and ROCK play critical roles in the regulation of stress fiber (SF) formation in cells. To determine whether $LT{\beta}R$ stimulation in fibroblastic reticular cells (FRCs) is involved in these signaling pathways, myosin light chain kinase inhibitor-7 (ML-7) was used to inhibit them. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic anti-$LT{\beta}R$ antibody-treated cells. The inhibition of ROCK activity with Y27632-induced changes in actin cytoskeleton organization and cell morphology in FRCs. Actin bundles rearranged into SFs, and phospho-myosin light chain (p-MLC) co-localized in FRCs. We checked the level of Rho-guanosine diphosphate (RhoGDP)/guanosine triphosphate (GTP) exchange activity using FRC lysate. When $LT{\beta}R$ was stimulated with agonistic anti-$LT{\beta}R$ antibodies, Rho-GDP/GTP exchange activity was markedly reduced. Regarding $LT{\beta}R$ signaling with a focus on MLCK inhibition, we showed that the phosphorylation of MLCs was reduced by $LT{\beta}R$ stimulation in FRCs. Cytoskeleton components, such as tubulin, b-actin, and phospho-ezrin proteins acting as membrane-cytoskeleton linkers, were produced in de-phosphorylation, and they reduced expression in agonistic anti-$LT{\beta}R$ antibody-treated FRCs. Collectively, the results suggested that MLCK and ROCK were simultaneously responsible for SF regulation triggered by $LT{\beta}R$ signaling in FRCs.

Effects of high Cell Density on growth-Associated Monoclonal Antibody Production by Hybridoma T0405 Cells Immobilized in Macroporous Cellulose carriers

  • Hideki Mochoda;Wang, Pi-Chao;Fr Jr. Nayve;Ryuji Sato;Minoru Harige;Nakao Nomura;Masatoshi Matsumura
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.5 no.2
    • /
    • pp.110-117
    • /
    • 2000
  • Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

  • PDF

In Vivo Tumor Cell Distribution of Antibody-Endostatin Fusion Protein for Tumor-Specific Targeting and Pharmacokinetics (암세포 표적지향화를 위한 항체-엔도스타틴 융합단백질의 체내동태 및 종양으로의 이행성)

  • Kang, Young-Sook;Lee, Na-Young
    • Journal of Pharmaceutical Investigation
    • /
    • v.33 no.4
    • /
    • pp.287-292
    • /
    • 2003
  • A novel antitumor agent, antibody-endostatin fusion protein $(anti-HER2/neu\;IgG3C_H3-Endostatin,\;AEFP)$ formed by genetic engineering procedure from antibody (Ab) which specifically targets to tumor cells ad angiogenesis inhibitor, endostatin (Endo) that has excellent antitumor effect, minimizes the toxicity of normal cells and selectively kills only tumor cells. The purpose of this study is to evaluate the phamacokinetic parameters and to analyze the localization of AEFP. After an intravenous injection of $150\;{\mu}l\;(5\;{\mu}Ci)\;[^{125}I]Ab,\;[^{125}I]AEFP$ to mice, blood was collected though retroorbital plexus from 15 min to 2880 min. Following the jugular vein injetion of $150\;{\mu}l\;(10\;{\mu}Ci)\;[^{125}I]Endo$, blood was collected by the use of carotid artery cannulation from 0.25 min to 30 min. Consequently, Endo was very rapidly removed from plasma compartment within 30 min. On the other hand, AEFP similar to Ab was slowly cleared from plasma. Also, Endo was metabolized about 40% within 30 min. However, AEFP was shown to metabolize less than 10% within 2880 min. The organ distribution of Endo was in order kidney, lung, spleen. Both Ab and AEFP were localized in order spleen, kidney, liver. Futhermore the tumor/blood distribution ratio of AEFP at 96 hours after injection is about 20 times higher than it of Endo at one hour after injection. In conclusion, these studies demonstrate that the anti-cancer or suppression of angiogenesis effect of Endo may be improved by the use of AEFP because the longer half life and stability of AEFP is able to selectively target antigens expressed on tumors.

Lab-on-a-Chip for Monitoring the Quality of Raw Milk

  • Choi Jeong-Woo;Kim Young-Kee;Kim Hee-Joo;Lee Woo-Chang;Seong Gi-Hun
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.8
    • /
    • pp.1229-1235
    • /
    • 2006
  • A lab-on-a-chip (LoC) was designed for simultaneous monitoring of microorganisms, antibiotic residues, somatic cells, and pH in raw milk. The LoC was fabricated from polydimethylsiloxane (PDMS) using microelectromechanical system (MEMS) technology, which consisted of two parts; a protein array and microchannel. The protein array was fabricated by immobilizing five types of antibodies corresponding to two microorganisms, two antibiotic residues, and somatic cells. A sol-gel film was deposited on a glass substrate to immobilize the antibodies. The target analytes in raw milk could be bound with the corresponding antibody by an immunoreaction, and the antigen-antibody complex was detected using fluorescence microscopy. SNARF-dextran was used as a pH indicator, and the SNARF-entrapped hydrogel was attached to the microchannel in the chip. After injecting the milk sample into the channel, the pH was measured by monitoring the change in fluorescence intensity by fluorescence microscopy. The on-chip simultaneous assay of two microorganisms (E. coli O157:H7 and Streptococcus agalactiae), two antibiotic residues (penicillin G and dihydrostreptomycin), and neutrophils was successfully accomplished using the proposed LoC system.

Different Immunology Mechanisms of Phellinus igniarius in Inhibiting Growth of Liver Cancer and Melanoma Cells

  • Zhou, Cui;Jiang, Song-Song;Wang, Cui-Yan;Li, Rong;Che, Hui-Lian
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.8
    • /
    • pp.3659-3665
    • /
    • 2014
  • To assess inhibition mechanisms of a Phellinus igniarius (PI) extract on cancer, C57BL/6 mice were orally treated with PI extractive after or before implanting H22 (hepatocellular carcinoma ) or B16 (melanoma) cells. Mice were orally gavaged with different doses of PI for 36 days 24h after introduction of H22 or B16 cells. Mice in another group were orally treated as above daily for 42 days and implanted with H22 cells on day 7. Then the T lymphocyte, antibody, cytokine, LAK, NK cell activity in spleen, tumor cell apoptosis status and tumor inhibition in related organs, as well as the expression of iNOS and PCNA in tumor tissue were examined. The PI extract could improve animal immunity as well as inhibit cancer cell growth and metastasis with a dose-response relationship. Notably, PI's regulation with the two kinds of tumor appeared to occur in different ways, since the antibody profile and tumor metastasis demonstrated variation between animals implanted with hepatocellular carcinoma and melanoma cells.

Cooperative Strategies and Swarm Behavior in Distributed Autonomous Robotic Systems Based on Artificial Immune System (인공 면역계 기반 자율분산로봇 시스템의 협조 전략과 군행동)

  • Sim, Kwee-Bo;Lee, Dong-Wook;Sun, Sang-Joon
    • Journal of Institute of Control, Robotics and Systems
    • /
    • v.6 no.12
    • /
    • pp.1079-1085
    • /
    • 2000
  • In this paper, we propose a method of cooperative control (T-cell modeling) and selection of group behavior strategy (B-cell modeling) based on immune system in distributed autonomous robotic system (DARS). An immune system is the living bodys self-protection and self-maintenance system. these features can be applied to decision making of the optimal swarm behavior in a dynamically changing environment. For applying immune system to DARS, a robot is regarded as a B-cell, each environmental condition as an antigen, a behavior strategy as an antibody, and control parameter as a T-cell, respectively. When the environmental condition (antigen) changes, a robot selects an appropriate behavior strategy (antibody). And its behavior strategy is stimulated and suppressed by other robots using communication (immune network). Finally, much stimulated strategy is adopted as a swarm behavior strategy. This control scheme is based on clonal selection and immune network hypothesis, and it is used for decision making of the optimal swarm strategy. Adaptation ability of the robot is enhanced by adding T-cell model as a control parameter in dynamic environments.

  • PDF

Detection of PCB77 by Indirect Competitive Enzyme-linked Immunosorbent Assay in Sea Sediment Samples

  • Chen, Han-Yu;Zhuang, Hui-Sheng;Yang, Guang-Xin
    • Bulletin of the Korean Chemical Society
    • /
    • v.34 no.3
    • /
    • pp.922-926
    • /
    • 2013
  • 3,3',4,4'-Tetrachlorobiphenyl (IUPAC PCB77) is one of seven indicative polychlorinated biphenyls (PCBs) in the surface sediments. The current study presents a novel polyclonal antibody for the determination of the PCB77 using indirect competitive enzyme-linked immunosorbent assay. Under optimum conditions, PCB77 was determined within the concentration range of 0.01-100 ${\mu}g\;L^{-1}$, with a detection limit of 0.057 ${\mu}g\;L^{-1}$. The assays were tested for their cross-reactivity profiles using 3 selected congeners and 4 Aroclor products. The assays were highly specific for coplanar PCB congeners, but less specific for Aroclor1248. The spiked recoveries from five sediment samples were 86%-114% for PCB77 from ELISA, which were satisfactory. The current study demonstrated that the developed antiserum and immunoassay procedure can be used to detect PCB77 in environmental samples. The results of the sediment analysis were confirmed by conventional GC/ECD.

Detection of Protein Molecules by Electrical Current Response Using Two-Electrode Method

  • Lyu, Hong-Kun;Woo, Sung-Ho;Han, Yoon-Soo;Lee, Hee-Ho;Shin, Jang-Kyoo
    • Journal of Sensor Science and Technology
    • /
    • v.20 no.2
    • /
    • pp.90-95
    • /
    • 2011
  • In order to protect human lives from disease, various biosensors having the potential to analyze a variety of biomolecules have been utilized. Biosensors constitute one of the most promising ways to monitor and detect various biomolecules corresponding to diseases. In this study, we demonstrate that the reaction of streptavidin molecules with biotin on a gold electrode can be detected using the twoelectrode method with a gold electrode and a platinum reference electrode. We also show the characteristics of the electrical current response. While detecting 2-${\mu}M$ streptavidin molecules dissolved in phosphate buffered saline(PBS) solution, we found that an analytical biosensor can operate on the principle of detecting an antigen-antibody reaction event of protein molecules using the two-electrode method. We think that the "potential step" method might be useful to detect the occurrence of any antigen-antibody reactions and can be combined with other devices or ICs such as BJTs, MOSFETs, and OP-amps for the detection of biomolecules of diseases.