What is it about?
During downstream processing of pharmaceutical proteins, purification of target protein with high effciency and yield is an important issue. This work is about the effect of different protein post-solubilization pH conditions on the efficacy of chromatographic processes for purification of recombinant streptokinase.
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Why is it important?
In the conventional purification of several pharmaceutical grade recombinant proteins, the pH value of the solubilized inclusion bodies may subjectively adjusted to values as low as 4.2 at the post-solubilization stage, and the clarified solution is subsequently utilized in the sequential chromatography process. We showed that chromatographic purification process done under such condition was not completely effective, as there was a significant target protein loss due to inadequate streptokinase purity (<90%) in most of the elution fractions. In this study, we showed that pH value of the solubilized inclusion bodies can have considerable impacts on the quality and quantity aspects of purification processes and significantly affects the yield and efficiency of large-scale purification of target protein. We demonstrated that post-solubilization higher pH (i.e., pH 6.0) could substantially improve the efficiency and yield of the chromatography procedures for purification of protein at large scale. This means that the complete final eluate (with 98% streptokinase purity and 91.4% recovery) obtained from the sequential purification process was fully exploitable for further active pharmaceutical ingredient (API) preparation.
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This page is a summary of: Effect of post-solubilization conditions on the yield and efficiency of recombinant streptokinase purification at large-scale, Protein Expression and Purification, September 2014, Elsevier,
DOI: 10.1016/j.pep.2014.06.007.
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