Journal of Blood Disorders & Transfusion

Purification of coagulation factor VIII by liquid chromatography

5^th World Hematologists Congress

August 18-19, 2016 London, UK

Elisabeth Cheng

Butantan Institute, Brazil

Posters & Accepted Abstracts: J Blood Disord Transfus

Abstract:

Factor VIII (FVIII) is an essential coagulation factor in the blood, which is missing or defective in patients with Hemophilia A, a life-threatening blood disorder affecting one in 5000 males. Currently, replacement therapy with FVIII concentrates constitutes the basis for hemophilia care. Because of the high costs and immunogenicity of the recombinant product, there is still a demand for the plasma derived concentrate, especially for the developing countries. Brazil has an excess of plasma generated as a byproduct of the production of red blood cell concentrates that can be used as source of fractionated plasma products. In the Process Development Laboratory at Butantan Institute, we have been working in the purification of FVIII based in a methodology that avoids the cryoprecipitate. Fresh frozen plasma bags are thawed, pooled and after adjusting pH, plasma is directly loaded to an anion exchange column, which is the method of choice to capture this protein, because it is cost effective, easily scalable and purifications result in high FVIII recovery. A second purification step for the separation of the vitamin-K dependent proteins, a family containing several coagulation factors and inhibitors has been the focus of our interest. Due to the difference in size, these proteins could be separated from FVIII on a gel filtration column. Furthermore, the higher histidine residue content on the surface of FVIII allowed the full separation on IMAC (immobilized metal affinity chromatography) column with good yield.

Biography :

Email: elisabeth.cheng@butantan.gov.br