Determination of recombinant hirudin structural deviants by capillary zone electrophoresis augmented with buffer additives

Add time:08/16/2019    Source:sciencedirect.com

The polypeptide hirudin is a potent and specific thrombin inhibitor used in anticoagulant therapy and naturally occurring in medicinal leech. Using gene-technology methods, recombinant (r) hirudin can be produced on a large scale. Purity evaluation of the synthesized r-hirudin is essential to monitor co-expressed structural deviants and degradation products before therapeutic use. Although the well established RP-HPLC analysis appears to be the method of choice, in the case of r-hirudin baseline separation of the structural deviants is not necessarily achieved. Capillary zone electrophoresis augmented with buffer additives was used as a complementary technique to separate r-hirudin successfully from several similar species, in order to provide characterization information, as well as performing purity control and stability studies.

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