Development of technology of blood coagulation factor VIII purification by the method of affinity chromatography on silica sorbents
Journal Title: Біологія тварин - Year 2016, Vol 18, Issue 2
Abstract
The manufacturing processes of chromatographic purification are used increasingly for plasma fractionation in the last few years. Implementation and combination of ion-exchange, affinity and size-exclusion chromatography have allowed the development of a new generation of therapeutic plasma derivatives, especially coagulation factors with improved purity and virus safety. The purpose of this work was to investigate the possibility of using the method of chromatographic purification of factor VIII on silica sorbents with triazine dyes as ligands. The paper presents the results of studies of purification blood coagulation factor VIII which is a combination of the method of fresh frozen plasma fractionation with the next step of affinity chromatography on macroporous silica sorbents. The main advantages and disadvantages of using triazine dyes as ligands are described. The suggested method contains a combination of previous precipitation of plasma proteins with barium citrate, adsorption with aluminium hydroxide, separation of precipitated proteins by centrifugation and using the method of negative affinity chromatography on the silica sorbents: Diasorb Active Bright-blue K, Diasorb Active-scarlet damask 4GT, Diasoorb-Procion Blue HB, Diasorb-Procion Gelb MXR. We have shown that the combination of previous fractionation of plasma with method of negative affinity chromatography on macroporous silica sorbents with synthetic dyes as ligands can achieve high purity factor VIII (about 100 times). For example, using sorbent Diasorb Active-scarlet damask 4GT increases the specific activity of factor VIII from 0.017±0.004 IU/mg to 1.940±0.023 IU/mg protein. This technology can be used along with other types of chromatography in the scheme of preparative receiving of factor VIII.
Authors and Affiliations
N. O. Shurko, T. V. Danysh
Keywords
Related Articles
- EP ID EP259221
- DOI 10.15407/animbiol18.02.152
- Views 124
- Downloads 0
Iodine aqua citrate impact on metabolic processes and productive qualities of piglets in nursery
The article presents the results of study on the iodine aqua citrate impact on metabolism and productive qualities in piglets from ablactation to intensive feeding (29–70 days). Complete feed with gramineous group of con...
The application of the RAPD-PCR method for identification of lactic acid bacteria
The heterogeneous properties of lactic acid bacteria (LAB) create a significant difficulties in their identification. The use of genotypic methods based on polymerase chain reaction (PCR) has opened new possibilities for...
Aquaculture objects DNA vaccination
Aquaculture, by making a powerful contribution to the production of products in the world, must be sustainable, ecological and economical. The growth of aquaculture volumes is inextricably linked with the outbreaks of in...
Dynamics of microstructural changes of white rat liver under the influence f e-selen in complex with Closaverm-A
The aim of the study was to examine the histostructural changes of liver at administration of E-selen in complex with Closaverm-A in therapeutic dose during 14 days. To this end, we used 48 white rats at the age of 2 3 m...
Location features of lymphoid tissue in immune formations of the small intestine and Meckel’s diverticulum walls in ducks
Mucous associated lymphoid tissue has a special place in the immune system, forming the first protective barrier against antigens that enter the body with food and air. In birds 70 % of lymphoid tissue that forms the par...