Viral inactivation and purification of active proteins

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...

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530412, 530413, 530414, 530415, 530427, A61K 3414, A61K 37475

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active

047497831

ABSTRACT:
Preparation of active therapeutic proteins comprising two essential steps. In the first step an active protein source is treated under conditions sufficient to inactivate any viruses present (e.g. via heat or chemical treatment). In the second step, the treated active protein source of moderate purity is then treated under conditions sufficient to remove impurities as well as denatured (once active) proteins resulting from the viral inactivation step. In an illustrative embodiment, a biologically active protein such as antithrombin is treated under conditions sufficient to assure viral inactivation and minimal antithrombin deactivation. After the viral inactivation, the antithrombin is contacted with immobilized heparin to complex only the active antithrombin which is then eluted by known means and processed further (e.g., to include desired excipients and then freeze dried). The preferred resulting concentrate is characterized by a high degree of purity, virus inactivation, and absence of denatured or inactive protein (generally less than about 10% by weight inactive form or, in the case of antithrombin, less than about 10% as determined, for example, by crossed immunoelectrophoresis).

REFERENCES:
patent: 4265924 (1981-05-01), Buhler et al.
patent: 4377514 (1983-03-01), Rubenstroth-Bauer et al.
patent: 4446134 (1984-05-01), Naito et al.
patent: 4673733 (1987-06-01), Chandra et al.

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