Large-scale purification of factor VIII by affinity chromatography: optimization of process parameters

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Booy, Marcel P.W.M. te and Faber, Anita and Jonge, Egge de and Wolterink, Ernst P. and Riethorst, Waander and Beugeling, Tom and Bantjes, Adriaan and Over, Jan and König, Boudewijn W. (1990) Large-scale purification of factor VIII by affinity chromatography: optimization of process parameters. Journal of Chromatography A, 503 . pp. 103-124. ISSN 0021-9673

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Abstract:The optimization of a new process for the extraction of human coagulation factor VIII (FVIII) from plasma with the tailor-made affinity matrix dimethylaminopropylcarbamylpentyl-Sepharose CL-4B (C3---C5 matrix) is described. First, plasma is applied to DEAE-Sephadex A-50 anion exchanger in order to separate a number of proteins, including coagulation factors II, IX and X (prothrombin complex), from FVIII. Subsequently, the unbound fraction of the ion exchanger, containing FVIII, is contacted with the C3---C5 affinity matrix. Optimization of the FVIII affinity chromatographic procedure is accomplished in terms of the ligand density of the matrix, adsorption mode (batch-wise versus column-wise adsorption and matrix to plasma ratio), and conditions of pH and conductivity to be applied on washing and desorption. In scale-up experiments, by processing 20 1 of plasma, the recovery (340 U VIII:C/kg plasma) and the specific activity (s.a.) (1.2 U VIII:C/mg protein) are better than those obtained by cryoprecipitation (recovery 300 U VIII:C/kg plasma, s.a. O.3 U VIII:C/mg protein). The newly developed process using the specially designed C3---C5 affinity matrix has potential application in the process-scale purification of FVIII.
Item Type:Article
Copyright:© 1990 Elsevier
Link to this item:http://purl.utwente.nl/publications/70635
Official URL:http://dx.doi.org/10.1016/S0021-9673(01)81494-1
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