Production Process
May be the best apotransferrin manufacturing method is the one patented and published by the Finish Red Cross.
According to it, the apotransferrin is produced from tested fraction IVpaste of the Cohn which is dissolved with water for injection. The protein solution is collected by filtration in the presence of filter aids is added and the precipitate is removed with a depth filter. EDTA is added to the solution . The clarified solution is applied to a chromatography column packed with SP Sepharose gel and pre-equilibrated with 40 m M sodium acetate buffer, pH 5·9. Other proteins and impurities are flushed away with the equilibration buffer. Transferrin is eluted with 40 m M Tris-HCl buffer, pH 7·5, and the pH of the collected transferrin-containing fraction is adjusted to pH 7·3 with the elution buffer. 1% (w/w) of Tween 80 and 0·3% (w/w) of tri-(n-butyl)phosphate are added for the solvent detergent (SD) treatment, and the solution is incubated at 26_ C for 10 h for inactivation of enveloped viruses. The partially purified apotransferrin solution is applied to a chromatography column packed with Q Sepharose gel pre-equilibrated with 40 m M Tris-HCl buffer, pH 7·5. The SD chemicals and protein impurities are flushed away with the equilibration buffer. Apotransferrin is eluted. The buffer of the protein solution is changed by diafiltration in an ultrafiltration unit, and the solution is concentrated to a protein concentration of 55 g/l. The pH of the pure apotransferrin solution is adjusted to pH 6-7. The solution is prefiltered for removal of viruses. The pure and virus-safe product is sterile filtered and aseptically filled into infusion bottles. The product is stored as a liquid formulation at refrigerator temperature or, in early development batches, optionally lyophilized.
Another production process on an industrial scale based on a simple and cost effective system is the one owned by Probitas Pharma when it bought the assets from Alpha Therapeutic Corporation in 2.003. This production process has also been patented.
Several other applications also involve the use of purified transferrin products. For instance, transferrin products are used as cell growth factors in serum-free cell culture media, or as reagents in diagnostic kits. You may get all these production processes in the patents page