Method for recombinant fibrinogen production

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification

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435 696, 530382, 530412, 530416, 530420, 530830, C07K 122, C07K 118, C07K 1475, C12P 2102

Patent

active

060374578

ABSTRACT:
The present invention provides methods of producing recombinant fibrinogen in a long-term mammalian cell culture system. Disclosed is a method for the production of recombinant fibrinogen, comprising: growing mammalian cells that express recombinant fibrinogen in a serum-free medium for a time of at least one month at a level of at least 5 .mu.g/ml; and then collecting portions of the conditioned medium at least twice during the culturing time of at least one month, with each portion containing at least 5 .mu.g/ml of recombinant fibrinogen. Also disclosed is a method for the production of recombinant fibrinogen, comprising: growing mammalian cells that express recombinant fibrinogen in a serum-free medium at a level greater than 1 .mu.g/ml; collecting at least a portion of the conditioned medium, and then purifying the fibrinogen from the medium by anion-exchange chromatography or affinity chromatography. In some embodiments of the invention, the medium is concentrated prior to the step of purifying the fibrinogen. In alternate embodiments, both the concentrating and purifying steps are carried out in the presence of at least one protease inhibitor.

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