Expression of a Protein A-Fucosyltransferase III fusion protein in insect cells

Vonach, B.; Hess, Bradley; Streift, M.; Leist, C.

doi:10.1007/978-94-011-5404-8_111

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“…In comparison with earlier work (Petri et al, 1997), where the High Five TM cells were cultivated in monolayers, the higher cell densities obtained by suspension cultures led also to a significantly higher product yield. This increased yield is in the order of magnitude of the data presented by Vonach et al (1997) for the production of the protein A fucosyltransferase III fusion protein using the same expression system. The much higher yields from High Five TM cells in comparison with Sf9 cells are probably due to the fact that High Five TM cells possess a more active and potent secretion apparatus.…”

Section: Discussionmentioning

confidence: 50%

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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: 50%