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High-Level Expression and Preparation of Recombinant Human Fibrinogen as Biopharmaceuticals.

08:00 EDT 15th October 2015 | BioPortfolio

Summary of "High-Level Expression and Preparation of Recombinant Human Fibrinogen as Biopharmaceuticals."

Fibrinogen is a large and complex glycoprotein containing two sets of each of three different chains (α, β, and γ). There have been no reports of high-level expression of fibrinogen at commercial levels using mammalian cultured cells such as CHO cells because of the difficulty in highly expressing a protein with such a complex structure.We achieved high-level (1.3 g/L or higher) expression of recombinant human fibrinogen using CHO DG44 cells by optimizing the expression system and culture conditions. We also succeeded in establishing a high-recovery preparation method for recombinant fibrinogen that rarely yields degraded products.To characterize the properties of the recombinant human fibrinogen, we performed SDS-PAGE; western blotting of the α, β, and γ chains using specific antibodies; and scanning electron microscopy (SEM) observations of fibrin fibers. We also evaluated the functional equivalence between recombinant fibrinogen and plasma fibrinogen with respect to the release of fibrinopeptides initiated by thrombin and its cross-linking properties. The basic properties of recombinant fibrinogen showed no apparent differences from those of plasma fibrinogen.Here we report the development of methods for the culture and preparation of recombinant human fibrinogen of satisfactory quality that can be scaled up to the commercial level.

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This article was published in the following journal.

Name: Journal of biochemistry
ISSN: 1756-2651
Pages:

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