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  • 1
    Electronic Resource
    Electronic Resource
    Purification, crystallization and preliminary X-ray crystallographic study of α-amylase from Bacillus stearothermophilus (2000)
    Copenhagen : International Union of Crystallography (IUCr)
    Acta crystallographica 56 (2000), S. 200-202 
    Details
    ISSN: 1399-0047
    Source: Crystallography Journals Online : IUCR Backfile Archive 1948-2001
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: A recombinant α-amylase from Bacillus stearothermophilus was found to be produced as several isoforms arising from different N-terminal processing. Some of those isoforms were purified to homogeneity and crystallized at 293 K using the hanging-drop vapour-diffusion method under the following conditions: 35 mM sodium acetate (pH 4.6), 6.25%(v/v) 2-propanol, in the presence of 1.23%(w/v) acarbose (a pseudo-oligosaccharide inhibitor) in the drop. The crystals diffracted beyond 2.0 Å resolution using synchrotron radiation at the Photon Factory, Tsukuba. They belong to the monoclinic space group P21, with unit-cell parameters a = 53.7 (2), b = 92.9 (4), c = 53.2 (2) Å, β = 109.4 (1)°.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1107/S0907444999015772
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  • 2
    Electronic Resource
    Electronic Resource
    Application of solvent extraction to ethanol fermentation (1984)
    Springer
    Applied microbiology and biotechnology 20 (1984), S. 371-377 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary The basic problems of applying solvent extraction to ethanol fermentation were investigated. The selection of solvents was based on the selectivity ratio, which was expressed as the ratio of the ethanol distribution coefficient to the water distribution coefficient. Solvents with high selectivity ratios of more than 50 were found mainly among the alcohols and esters. However, most of these solvents were toxic to ethanol-producing microorganisms. We tried to make a barrier to solvent molecules beneath the surface of gel beads immobilizing the cells as a protection against solvent toxicity. Porapack Q was found to be an effective barrier, and the ethanol production rate of immobilized cells protected with Porapack Q did not change event after the production of eight batches in medium saturated with sec-octanol, which was the most toxic solvent used in our experiments.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00261937
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  • 3
    Electronic Resource
    Electronic Resource
    In vitro induction of HLA-A2402-restricted and carcinoembryonic-antigen-specific cytotoxic T lymphocytes on fixed autologous peripheral blood cells (1998)
    Springer
    Cancer immunology immunotherapy 47 (1998), S. 90-96 
    Details
    ISSN: 1432-0851
    Keywords: Key words Carcinoembryonic antigen ; Cytotoxic T lymphocyte ; HLA-A2402 ; Epitope peptide ; Immunotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract HLA-A2402-restricted and carcinoembryonic-antigen(CEA)-specific cytotoxic T lymphocytes (CTL) were induced by culturing human peripheral blood mononuclear cells (PBMC) on formalin-fixed autologous adhesive PBMC that had been loaded with CEA-bound latex beads. The CTL killed the CEA-producing HLA-type matched cancer cells, but not the non-producers of CEA, at an effector/target ratio of 10 within 24 h. On the basis of available HLA-A24-binding peptides, we have also attempted to identify the epitope peptide recognized by the CTL. The peptide CEA652(9), TYACFVSNL, stimulated the CTL most strongly when pulsed on HLA-A2402-expressing target cells. The other nine peptides so far tested were also active, but less efficient in their effect on CTL. The CTL failed to kill target cells pulsed with the HLA-A2-binding CEA peptide, CAP-1. The CTL were also generated on the fixed adherent cells previously pulsed with the peptide CEA652(9). Cytotoxic activity of the CTL was inhibited by monoclonal antibodies against CD3, CD8, and MHC class I molecules. These results suggest that human autologous CTL will be inducible on the autologous fixed PBMC without use of the cultured target cancer cells if tumor antigenic protein is available.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002620050508
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  • 4
    Electronic Resource
    Electronic Resource
    Eligibility of antigenic-peptide-pre-loaded and fixed adhesive peripheral blood cells for induction of cytotoxic T lymphocytes from cancer patients with elevated serum levels of carcinoembryonic antigen (2000)
    Springer
    Journal of cancer research and clinical oncology 126 (2000), S. 383-390 
    Details
    ISSN: 1432-1335
    Keywords: Key words Carcinoembryonic antigen ; Cytotoxic T lymphocyte ; HLA-A2402 ; Epitope peptide ; Immunotherapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The inducibility of cytotoxic T lymphocytes (CTL) that react with carcinoembryonic antigen (CEA) was tested in cancer patients with elevated (more than 5 ng/ml) serum CEA levels when antigen presentation was carried out with paraformaldehyde-fixed adhesive peripheral blood mononuclear cells (PBMC) from the patient that had been pre-loaded with CEA652(9), an HLA-A2402-restricted tumor antigenic peptide derived from CEA. By culturing fresh autologous PBMC on the fixed cell layer in medium containing interleukin-1, -2, -4 and -6, three out of eight patients developed CTL. The CTL from two of these patients killed CEA-protein-producing gastric cancer cells carrying HLA-A2402 and the cells from the remaining patient killed CEA-non-producing stomach cancer cells pre-loaded with CEA652(9). The results suggest that a single antigenic peptide on the fixed adhesive cells will allow the ex vivo induction of peptide-reactive CTL that are easier to handle and allow antigen presentation without tedious preculture of the “professional” antigen-presenting dendritic-cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00008486
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  • 5
    Electronic Resource
    Electronic Resource
    Construction of glomerular epithelial cells in vitro for removal of immune complex (1999)
    Springer
    Journal of artificial organs 2 (1999), S. 170-175 
    Details
    ISSN: 1619-0904
    Keywords: Remove immune complex ; Glomerular epithelial cell ; CR1 expression ; DNA/anti-DNA antibody ; Phagocytosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Technology
    Notes: Abstract Patients with systemic lupus erythematosus (SLE), an autoimmune disease caused by excessive amounts of immune complex in the serum, have a loss of complement receptor type 1 (CR1), a receptor of complements C3b and C4b, on glomerular epithelial cells. In order to calrify the biological function of CR1 on renal glomeruli and the correlation with clearance of immune complex, a full length of human tonsil CR1 cDNA was transfected to normal rat glomerular epithelial cells (SGE1) to enable CR1 expression in the long term (more than 1 year). As compared with the non-CR1-expressing cells, the CR1-expressing cells showed a higher binding effect to immune complex in 60 min, and the binding effect was mediated by complement. Blocking of the binding effect by anti-CR1 antibody indicates that CR1 plays an important role in removal of immune complex. Results from microscopy showing that the constructed cell has an enhanced phagocytic ability in the presence of complement suggest that the mechanism of removing immune complex by CR1-expressing cells has three steps: opsonization of immune complex by complement, the ligand-receptor interaction between opsonized complex and CR1 on the cell, and phagocytosis of complex by CR1-expressing cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02480062
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  • 6
    Electronic Resource
    Electronic Resource
    Succinoyl trehalose lipid induced differentiation of human monocytoid leukemic cell line U937 into monocyte-macrophages (1995)
    Springer
    Cytotechnology 19 (1995), S. 79-88 
    Details
    ISSN: 1573-0778
    Keywords: trehalose lipid ; U937 ; monocytic differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A novel type of succinoyl trehalose lipid (STL-1) prepared from n-hexadecane-culture ofRhodococcus erythropolis SD-74 markedly inhibited the growth of a human monocytoid leukemic cell line, U937, and induced its morphological alteration along a monocyte-macrophage lineage. STL-1 markedly increased differentiation-associated characteristics in macrophage, such as nitroblue tetrazolium reducing ability, appearance of Fc receptor, phagocytic activities in U937. Furthermore, U937 cells, which were activated with STL-1 exhibited cytotoxic activity against human lung carcinoma cell line A549. However, STL-1 did not affect growth of a normal human fetal lung cell line TIG-1. The individual components of STL-1, neither sugar moiety nor fatty acids in the free form, were effective at inducing the differentiation of U937 cell. From these results, we concluded that STL-1 has low cytotoxicity against normal human cells and the ester molecule itself is responsible for the activity of inducing differentiation of human monocytoid leukemic cell line U937 into monocyte-macrophage which results in the stimulation of the production of some cytotoxic substances.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749758
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  • 7
    Electronic Resource
    Electronic Resource
    Physical and biological studies on DNA/anti-DNA immune complex formed in vitro (1997)
    Springer
    Cytotechnology 25 (1997), S. 165-171 
    Details
    ISSN: 1573-0778
    Keywords: anti-DNA-antibody ; complement receptor ; immune complex ; NZB/W mice ; SLE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In vitro preparation of DNA/anti-DNA immune complex using monoclonal antibody and low molecular weight homogeneous DNA was described and the characteristics of the immune complex were identified. The immune complex was formed by the monoclonal antibody with DNA effectively at high ratios of antibody/DNA. The activation and binding ability of the immune complex to the complement was considerably high, whereas the capacity to bind red blood cells via C3b complement component receptor was shown relatively low. The assay of sucrose density gradient ultracentrifugation indicated that the sedimentation coefficient of the immune complex was between 10S and 23S. Studies of organ distribution and uptake of IC demonstrated a particular affinity to the kidney tissue. The significance of these results with respect to the latent pathogenicity of the immune complex was discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007926809023
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  • 8
    Electronic Resource
    Electronic Resource
    Adaptation of hybridoma cells to higher ammonia concentration (1991)
    Springer
    Cytotechnology 7 (1991), S. 103-112 
    Details
    ISSN: 1573-0778
    Keywords: adaptation ; ammonia ; hybridoma ; continuous culture ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Using two mouse-mouse hybridoma cell lines, the response to ammonia step and serial changes was investigated in batch and continuous cultures with serum-free medium. The inhibitory effect of ammonia on cell growth depended on the cultivation mode, and differed markedly between cell lines. The cell line, 4C10B6 producing IgG monoclonal antibody against Pseudomonas, showed a high adaptation ability to ammonia. The 4C10B6 cells could grow under ammonia concentration as high as 21 mmol/l NH4Cl with a viability of 80% in the continuous culture with serial increase in ammonia concentration. Whereas, in the batch culture with ammonia step change the cell growth completely ceased at 12 mmol/l NH4Cl. The other cell line, TO-405 producing IgG monoclonal antibody against hepatitis B surface antigen, could not adapt to ammonia, and the cell growth did not occur at 9 mmol/l NH4Cl even under the ammonia serial change.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350916
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  • 9
    Electronic Resource
    Electronic Resource
    Effects of ammonium ion removal on growth and MAb production of hybridoma cells (1995)
    Springer
    Cytotechnology 18 (1995), S. 35-50 
    Details
    ISSN: 1573-0778
    Keywords: ammonium removal ; cross-flow filtration ; monoclonal antibody ; membrane fouling ; perfusion culture ; zeolite ; hybridoma cells ; defined medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Production of monoclonal antibody against hepatitis B surface antigen was carried out by perfusion culture coupled with a selective removal system for ammonium ion. The removal system is composed of three sub-systems namely, cell separation by cross-flow ceramic filter, dialysis by hollow fiber module and ion-exchange by zeolite A-3 packed bed column. The ammonium ion concentration in the culture broth was effectively maintained below the inhibitory level, and the viable cell density reached 2.5×107 cells ml−1 which was three times that of conventional perfusion cultures. The monoclonal antibody accumulated to a concentration as high as 26.3×105 mIU−1. This is already almost half of the amount producedin vivo. The numerical investigation of the ammonium ion removal system showed the possibility to improve much more the performance of this perfusion cultivation system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00744318
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  • 10
    Electronic Resource
    Electronic Resource
    Selective removal of ammonia from animal cell culture broth (1991)
    Springer
    Cytotechnology 6 (1991), S. 121-130 
    Details
    ISSN: 1573-0778
    Keywords: ammonia removal ; hybridoma ; HBs monoclonal antibody ; zeolite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Serum-free perfusion cultures of hybridoma TO-405 cells were carried out in spinner flasks coupled with zeolite A-3 packed beads. Ammonia was selectively removed from the culture broth by passing cell free permeate from ceramic cross flow filtration, through the zeolite packed bed. Ammonia concentration in the culture broth was effectively maintained between 1 to 4 mmol/l which was below the inhibitory concentration for cell growth. Maximum cell density levels of 107 cells/ml as well as improved percentage cell viability higher than in serum-supplemented cultures were feasible in this system. The possible effects of shear stress, generated by variation of the flow rates of the broth through the ceramic filter module, on the growth of the hybridoma cells were investigated. Backwashing, by reversing the direction of the permeate, was found necessary to prolong the life of the filter. Variation of the flow rates of the broth through the ceramic module between 0.29 m/s to 0.59 m/s did not cause immediate cell damage but growth was repressed at the higher flow rate. This study also showed that glutamine appears to be one of the factors limiting the growth of the hybridoma cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00373029
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