Characterization of growth and recombinant protein production of the insect cell line BTI-Tn-5Bl-4 (High Five™) in suspension culture

Chico, Ernesto; Kobold, Anja; Haase, Bernd; Jäger, Volker

doi:10.1007/978-94-011-5404-8_120

Cited by 1 publication

(3 citation statements)

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“…Here only a feeding of glucose together with the amino acids asparagine and glutamine, increased the triabin yield in the process. Chico et al (1997) had also described for High Five TM cells that these two amino acids were limiting substrates in other commercially available media. Asparagine and glutamine possess a double importance for growth and virus replication, because they not only serve as protein building blocks but also because they can be used to provide energy.…”

Section: Discussionmentioning

confidence: 99%

“…The BTI-Tn5B1-4 (High Five TM ) insect cell line has gained widespread use within baculovirus/insect cell expression system because many secreted recombinant proteins are produced at considerably higher rates than in Spodoptera frugiperda derived cell lines (e.g. Wickham et al, 1992;Davis et al, 1993;Wickham and Nemerow, 1993;Chico et al, 1997). Recently this has also been confirmed for the novel thrombin inhibitor triabin (Petri et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, we have extended these studies with suspension adapted High Five TM cells. As it is well known that the productivity of the baculovirus/insect cell expression system is strongly influenced by (1) the state of the host cell, (2) the multiplicity of infection, (3) the cell density at the time of infection as well as by (4) nutrient and (5) oxygen supply (Reid et al, 1995;Kioukia et al, 1995;Chico et al, 1997) experiments for the optimization of the triabin production were performed and are described here.…”

Section: Introductionmentioning

confidence: 99%

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Vallazza¹,

Petri²

1999

Cytotechnology

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The isolation of a new type of thrombin inhibitor, called triabin, from the saliva of the hematophagous bug Triatoma pallidipennis, has recently been described. In the in vitro platelet aggregation inhibition assay triabin has a similar potency as the thrombin inhibitor hirudin now in phase III clinical trials. However, in another in vitro assay using a low molecular weight substrate for thrombin, triabin does not inhibit thrombin completely even at 6 fold higher molar doses in comparison with hirudin. This means that triabin has a novel mode of action towards thrombin making triabin into an interesting candidate as a therapeutic agent. Recently it has been shown that a recombinant baculovirus can be efficiently used for the triabin production in insect cells and that the yields in adherent cultures of High Five™ cells (approx. 20 mg l-1) were about 7 fold higher than in adherent cultures of Sf9 cells (approx. 3 mg l- 1). To optimize the triabin yield from the baculovirus/insect cell expression system, experiments were performed with suspension adapted cultures of High Five™ cells to investigate the effects of the state of the host cell, of the multiplicity of infection, of the cell density at the time of infection and of supplementation of the medium with nutrients and oxygen. Triabin yields of up to 200 mg l-1, as determined by an activity assay, could finally be obtained here.

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Section: Discussionmentioning

confidence: 99%