Determination of recombinant hirudin structural deviants by capillary zone electrophoresis augmented with buffer additives.
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.
Dönges
Reiner
R
Aventis Behring, Quality Control, 35002 Marburg, Germany. reiner.doenges@aventis.com
Brazel
Dieter
D
D000595:Amino Acid Sequence; D002021:Buffers; D019075:Electrophoresis, Capillary / Q000379:methods*; D006629:Hirudins / Q000032:analysis* / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry
D002021:Buffers; D006629:Hirudins; D011994:Recombinant Proteins