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:: Volume 2, Issue 4 (Summer 2005) ::
Sci J Iran Blood Transfus Organ 2005, 2(4): 91-98 Back to browse issues page
Improving purification of coagulation F IX using heparin affinity chromatography and its comparison with ion exchange chromatography
S.Sh. Musavi Motlagh , H. Rezvan , A.A. Pourfathollah , M.K. Mousavi Hosseini
Abstract:   (18540 Views)

Abstract

 

Background and Objectives

Hemophilia B is a genetic disorder due to deficiency or complete absence of factor IX coagulation factor. Treatment of choice for these patients is use of factor IX concentrates. Therefore, purification of plasma proteins and seperation of factor IX have been major objectives for scientists involved in this field. In this respect, purification procedure using ion exchange chromatography is widely used, but in the past decade affinity chromatography was also introduced. The objective of the present study has been to apply both techniques for the purification of factor IX and compare the quality and yield of the product.

 

Materials and Methods

For the purification procedure, chromatography columns (XK-16), containing DEAE sepharose and Heparin sepharose were used. Factor IX coagulation activity was measured using a one-stage coagulation assay and factor IX antigen was quantified using ELISA technique.

 

Results

The specific activity and relative increase in purity of factor IX was calculated and it was demonstrated that specific activity improved from 3.1 IU/mg using DEAE ion exchange to 29 IU/mg when affinity chromatography was added and purity was increased from 155 to 1450 respectively.

 

Conclusions

The present study demonstrates that addition of an affinity chromatography step using heparin sepharose is a major improvement in the purification of factor IX, where both specific activity and purity are increased considerably.

 

 

Key words:  Heparin affinity chromatography, Factor IX, Hemophilia B, Ion exchange chromatography

Keywords: Heparin affinity chromatography, Factor IX, Hemophilia B, Ion exchange
Full-Text [PDF 274 kb]   (2906 Downloads)    
Type of Study: Research | Subject: General
Published: 2014/06/30
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Musavi Motlagh S, Rezvan H, Pourfathollah A, Mousavi Hosseini M. Improving purification of coagulation F IX using heparin affinity chromatography and its comparison with ion exchange chromatography. Sci J Iran Blood Transfus Organ 2005; 2 (4) :91-98
URL: http://bloodjournal.ir/article-1-31-en.html


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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 2, Issue 4 (Summer 2005) Back to browse issues page
فصلنامه پژوهشی خون Scientific Journal of Iran Blood Transfus Organ
The Scientific Journal of Iranian Blood Transfusion Organization - Copyright 2006 by IBTO
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