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  • Articles  (25)
  • monoclonal antibody  (23)
  • Lepidoptera
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  • Springer  (25)
  • 1990-1994  (25)
  • Process Engineering, Biotechnology, Nutrition Technology  (25)
  • 1
    Electronic Resource
    Electronic Resource
    A serum-free medium for hybridoma cell culture (1993)
    Springer
    Cytotechnology 11 (1993), S. 169-174 
    Details
    ISSN: 1573-0778
    Keywords: cell culture ; hybridoma ; monoclonal antibody ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The effects of several different substances, including insulin, transferrin, ethanolamine, selenite and butyrate on the growth of murine hybridoma 2F7 cells, which secrete monoclonal antibody against small cell lung cancer, were investigated, and a serum-free medium SFMI was formulated. The effects of taurine, spermidine, progesterone and adenine on the cell growth were tested further on the basis of the medium SFMI, and a modified serum-free medium SFM II was established. On the basis of medium SFM II, the substitution tests of ferric citrate for transferrin were carried out, and it was found that transferrin could be replaced. The experiments suggested that the formulated serum-free medium was suitable for 2F7 cell growth and monoclonal antibody secretion, and thus facilitated subsequent purification of monoclonal antibody.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749866
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  • 2
    Electronic Resource
    Electronic Resource
    Electron microscopy of hybridoma cells with special regard to monoclonal antibody production (1990)
    Springer
    Cytotechnology 4 (1990), S. 13-28 
    Details
    ISSN: 1573-0778
    Keywords: monoclonal antibody ; hybridoma ; electron microscopy ; endoplasmic reticulum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Electron microscopy of mouse hybridoma cell lines shows that the major difference between non, low and high producer cell lines is the amount of endoplasmic reticulum. Vesicular-tubular or cavernous structures of endoplasmic reticulum, which can survive long after cell death, are particularly abundant in producer cell lines. Immunogold labelling with anti-mouse IgG reveals that antibodies are predominantly located in these structures. The cell membrane undergoes structural changes during the late stages of batch culture with the disappearance of microvilli and the appearance of blebs and deep indentations. Necrosis disrupts the cytoplasmic structures and the nucleus is last to degrade.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00148807
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  • 3
    Electronic Resource
    Electronic Resource
    Antibody production in packed bed reactors using serum-free and protein-free medium (1990)
    Springer
    Cytotechnology 4 (1990), S. 279-283 
    Details
    ISSN: 1573-0778
    Keywords: animal cell culture ; hybridoma ; monoclonal antibody ; packed bed reactor ; continuous culture ; perfusion ; protein-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The present work demonstrates the utility of packed bed reactors for the production of monoclonal antibody. We present data from a continuous process run for the production of over 100 grams of antibody, using serum-free medium. An additional pilot run also demonstrates the potential for continued antibody production under protein-free conditions, using a standard basal medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00563788
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  • 4
    Electronic Resource
    Electronic Resource
    Cell cycle kinetics of the accumulation of heavy and light chain immunoglobulin proteins in a mouse hybridoma cell line (1994)
    Springer
    Cytotechnology 14 (1994), S. 205-218 
    Details
    ISSN: 1573-0778
    Keywords: Cell cycle ; flow cytometry ; heavy chain ; hybridoma ; light chain ; monoclonal antibody ; population balance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Rates of accumulation of immunoglobulin proteins have been determined using flow cytometry and population balance equations for exponentially growing murine hybridoma cells in the individual G1, S and G2+M cell cycle phases. A producer cell line that secretes monoclonal antibodies, and a nonproducer clone that synthesizes only κ-light chains were analyzed. The pattern for the kinetics of total intracellular antibody accumulation during the cell cycle is very similar to the previously described pattern for total protein accumulation (Kromenaker & Srienc 1991). The relative mean rate of heavy chain accumulation during the S phase was approximately half the relative mean rate of light chain accumulation during this cell cycle phase. This indicates an unbalanced synthesis of heavy and light chains that becomes most pronounced during this cell cycle phase. The nonproducer cells have on average an intracellular light chain content that is 42% lower than that of the producer cells. The nonproducer cells in the G1 phase with low light chain content did not have a significantly higher rate of light chain accumulation relative to other G1 phase nonproducer cells. This is in sharp contrast to what was observed for the G1 phase producer cells. In addition, although the relative mean rate of accumulation of light chain was negative for G2+M phase nonproducer cells, the magnitude of this relative mean rate was less than half that observed for the producer cells in this cell cycle phase. This suggests that the mechanisms that regulate the transport of fully assembled antibody molecules through the secretion pathway differ from those which regulate the secretion of free light chains. The results reported here indicate that there is a distinct pattern for the cell cycle dynamics of antibody synthesis and secretion in hybridomas. These results are consistent with a model for the dynamics of secretion which suggests that the rate of accumulation of secreted proteins will be greatest for newborn cells due to an interruption of the secretion pathway during mitosis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749617
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  • 5
    Electronic Resource
    Electronic Resource
    Methods and strategies available for the process control and optimization of monoclonal antibody production (1994)
    Springer
    Cytotechnology 14 (1994), S. 219-232 
    Details
    ISSN: 1573-0778
    Keywords: animal cells ; cellular metabolism ; cultivation ; hybridomas ; modelling ; monoclonal antibody ; process control ; optimisation ; simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The objective of this paper is to explore the range of methods and strategies available for the process control and optimization of monoclonal antibody production by hybridoma cell culture. Emphasis will be placed on the choice of the level of complexity incorporated into the process control and optimisation procedure. It will be shown that the behaviour of hybridomas in culture is influenced by sophisticated cellular metabolic activities and various interactive environmental factors and that the understanding and modelling of the way hybridomas grow in the bioreactor should enable optimisation of bioreactor operating conditions to achieve maximum monoclonal antibody formation. However, due to the lack of on-line instrumentation of important biological variables and the incomplete knowledge of hybridoma cultivation process, there exist many limitations and challenges to the advent of applications of process control and optimisation in this field. To solve the problem, introduction of industrially practical biological measurements and development of new control concepts are inevitable. At the end of this paper, we shall discuss possible schemes for the control of the physsiological state of cells in order that balanced cell growth and maximum monoclonal antibody synthesis may be achieved.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749618
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  • 6
    Electronic Resource
    Electronic Resource
    Weaning of three hybridoma cell lines to serum free low protein medium (1991)
    Springer
    Cytotechnology 6 (1991), S. 65-78 
    Details
    ISSN: 1573-0778
    Keywords: hybridoma ; monoclonal antibody ; serum free culture ; low protein medium ; weaning protocol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A general weaning procedure is described which allowed a range of hybridomas to be weaned readily off serum without loss of antibody production. Initial work was carried out with one cell line only (SPO1 cells) and one serum substitute containing a final protein concentration of 40 mg l-1. The SPO1 cells were first adapted to a range of readily available basal media and then weaned off serum by a range of protocols. From this work an optimal weaning protocol and basal medium for weaning were determined. These were then used to wean the SPO1 cells and two other cell lines off serum with a second, protein free, serum substitute with varying concentrations of defined proteins added. All three cell lines investigated were readily weaned off serum by this protocol at protein concentrations as low as 1 mg l-1. No loss of antibody production was observed with any of the cell lines. The weaning procedure outlined is both simple and rapid and has been successfully adopted in our laboratory by relatively inexperienced cell culture technicians.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00353704
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  • 7
    Electronic Resource
    Electronic Resource
    Establishment and characterization of monoclonal antibodies against Chuzan virus K-47 (1991)
    Springer
    Cytotechnology 6 (1991), S. 13-21 
    Details
    ISSN: 1573-0778
    Keywords: Chuzan virus ; monoclonal antibody ; neutralizing antibody ; forward sandwich assay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We established sixteen mouse monoclonal antibodies reactive to Chuzan virus K-47 strain using P3-X63-Ag8-U1 cells as fusion partner cells. Among them, CG53/2/4 recognized a 100K structural protein of the virus. The 100K antigen lost it's antigenicity for CG53/2/4 after mild periodate oxidation treatment, suggesting that the 100K viral antigen is a glycoprotein. In addition, CG53/2/4 neutralized the viral infectivity. This indicates that the 100K glycoprotein is essential for the infection of the virus. The other monoclonal antibodies reacted with a 41K antigen of the virus. Especially CG1/1 showed the highest reactivity to the virus. Forward step sandwich assay using CG1/1 and biotinylated CG53/2/4 could detect the virus at 10TCID50/ml. Therefore, these monoclonal antibodies can evantually predict the virus infection to the animals before their sideration.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00353698
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  • 8
    Electronic Resource
    Electronic Resource
    Serum-free medium for murine and human lymphoid and hybridoma cell lines (1991)
    Springer
    Cytotechnology 6 (1991), S. 33-38 
    Details
    ISSN: 1573-0778
    Keywords: hybridoma ; monoclonal antibody ; myeloma ; serum-free and tissue plasminogen activator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We established a serum-free medium of low protein content(125μg/ml) TYI 100, consisting of three hormones and five growth factors for the growth of lymphoid and hybridoma cell lines. In TYI 100 medium, mouse and human hybridomas grew equally well as in RPMI 1640 supplemented with 10% fetal bovine serum (10% FBS) without adaptation to the serum-free medium. TYI 100 medium allowed several passages of mouse hybridoma lines and the total cell number was more than in 10% FBS. TYI 100 medium also supported growth of myelomas and anchorage dependent cell lines, Bowes and CHO, well. TYI 100 medium is composed of inexpensive supplements and is therefor applicable to large scale culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00353700
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  • 9
    Electronic Resource
    Electronic Resource
    Improvement in the proliferative activity of human-human hybridomas at low cell density by transfection with bFGF gene (1991)
    Springer
    Cytotechnology 6 (1991), S. 93-103 
    Details
    ISSN: 1573-0778
    Keywords: cell improvement ; FGF ; hepatitis B virus ; human-human hybridoma ; monoclonal antibody ; proliferative activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Highly purified recombiaant basic fibroblast growth factor (rbFGF) and acidic FGF (aFGF) stimulated the proliferation of human-human (h-h) hybridomas to the extent of over four-fold from a low cell density such as 1×103 cells per ml in a serum-free medium in 24-well plates. The stimulatory effect of rbFGF was also observed in various lymphoid cell lines. Expecting that FGF could be an autocrine growth factor, we introduced bFGF gene into a h-h hybridoma using an expression plasmid induced by dexamethasone. The transformed cells thus obtained, HPO-75.11bbFGF-7, were able to grow well from a low inoculum density in a serum-free medium and antibody production was also increased when bFGF gene expression was induced. The transformed cells could grow at clonal density in a serum-free medium in 96-well plates, though the original cells could not. We also obtained a more practical transfectant, HPO-75.29-H74, using a high-shear stress adapted clone as the recipient and an expression plasmid having bFGF gene under the control of metallothioneine-I promoter. The HOP-75.29-H74 cells were capable of growing and producing human monoclonal antibody against hepatitis B virus surface antigen from an inoculum density of 1×103 cells per ml in an agitation vessel without addition of an inducer.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00373026
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  • 10
    Electronic Resource
    Electronic Resource
    High density culture of hybridoma cells using a perfusion culture vessel with an external centrifuge (1990)
    Springer
    Cytotechnology 3 (1990), S. 239-244 
    Details
    ISSN: 1573-0778
    Keywords: hybridoma ; perfusion culture ; monoclonal antibody ; serum-free culture ; centrifuge
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The influence of centrifugal force on the growth of cells was examined by exposing the cells of the mouse-human hybridoma X87 line to centrifugal force (100–500 G) for ten minutes twice a day and comparing the static culture with that of unexposed cells. In this experiment, both cell proliferation and specific antibody productivity were independent of the centrifugal effect, and gave the same results as in the case of no exposure to centrifugal force. High density cultivation of the mouse-human hybridoma X87 line was obtained by a perfusion system where the cells were separated from the culture medium by continuous centrifugation. In the serum-free culture, the maximum viable cell density exceeded 107 cells/ml, and monoclonal antibody was stably produced for 37 days. The results in this culture were equivalent to those obtained by intermittent centrifugal cell separation from the culture medium, and separation by gravitational settlement.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00365487
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