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. 2009 Apr;1(1):47-51.

Production and purification of streptokinase by protected affinity chromatography

Mohammad Babashamsi  1 , Mohammad Hossein Razavian, Mohammad Reza Nejadmoghaddam
Affiliations

Production and purification of streptokinase by protected affinity chromatography

Mohammad Babashamsi et al. Avicenna J Med Biotechnol. 2009 Apr.

Abstract

Streptokinase is an extracellular protein, extracted from certain strains of beta hemolytic streptococcus. It is a non-protease plasminogen activator that activates plasminogen to plasmin, the enzyme that degrades fibrin cloth through its specific lysine binding site; it is used therefore as a drug in thrombolytic therapy. The rate of bacterial growth and streptokinase production was studied in condition of excess glucose addition to culture media and its pH maintenance. The streptokinase product of the bacterial culture was preliminary extracted by salt precipitation and then purified by affinity chromatography on plasminogen substituted sepharose-4B in a condition that the plasminogen active site was protected from streptokinase-induced activation. The purity of streptokinase was confirmed by SDS-PAGE and its biological activity determined in a specific streptokinase assay. The results showed that in the fed-batch culture, the rate of streptokinase production increased over two times as compared with the batch culture while at the same time, shortening the streptokinase purification to a single step increased the yield over 95% at the chromatography stage.

Keywords: Affinity chromatography; Culture; Plasminogen; Purification; Streptokinase.

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Figures

Figure 1
Figure 1
Evaluation of plasminogen purification by SDS-PAGE (Left to right): 1. MW marker 2. Elution dialyzed 3. Washing with 0.1 M phosphate buffer 4. Human plasma
Figure 2
Figure 2
Variation of streptokinase [production rate] in culture media by pH regulation and glucose addition
Figure 3
Figure 3
Evaluation of streptokinase purification by SDS-PAGE (Left to right): 1- MW marker 2- Ammonium sulfate extract 3-Purified and dialyzed streptokinase 4-BSA
See this image and copyright information in PMC

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