Osteoinductive protein mixtures and purification processes

Details Osteoinductive protein mixtures and purification processes Osteoinductive protein mixtures and purification processes

A - Human Necessities

61

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A61K 38/18 (2006.01) A61F 2/28 (2006.01) A61K 35/32 (2006.01) A61K 38/02 (2006.01) A61L 27/02 (2006.01) A61L 27/22 (2006.01) C07K 1/16 (2006.01) C07K 1/18 (2006.01) C07K 14/51 (2006.01) A61F 2/00 (2006.01) A61F 2/02 (2006.01) A61F 2/32 (2006.01) A61F 2/38 (2006.01) A61F 2/44 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2107481

A method for purifying bone-derived osteoinductive factors including an ultrafiltration process, an anion exchange pro- cess, a cation exchange process, and a reverse phase HPLC process. The ultrafiltration process preferably includes a first ultrafil- tration step using a membrane having a nominal molecular weight cutoff of approximately 100 kilodaltons (kD) and a second ul- trafiltration step employing a membrane having a nominal molecular weight cutoff of approximately 10 kD. For the anion exchange process, a strongly cationic resin is used, preferably having quaternary amine functional groups. Typically, the eluant for the anion exchange process has a conductivity from about 10,260 micromhos (µmhos) (1.026 x 10-2 siemens (S)) to about 11;200 µmhos (1:120 x 10-2S). For the ration exchange process, a strongly anionic resin is used, preferably having sulfonic acid functional groups. The eluant for the ration exchange process typically has a conductivity from about 39,100 µmhos (3.91 x 10-2S) to about 52;700 µmhos (8:27 x 10-2S) or more. The HPLC process typically utilizes a column containing hydro- carbon-modified silica packing material. The osteoinductive proteins can be eluted from the HPLC column with an acetoni- trile solution in combination with aqueous trifluoracetic acid. The purification processes yield osteoinductively active pro- tein mixtures.

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