Separation of r-hirudin from similar substances by capillary electrophoresis.
The thrombin inhibitor r-hirudin is a peptide of 65 amino acids and with a pI of 4.4. Capillary electrophoresis (CE) was used to separate r-hirudin from seven closely related substances which may be found as by-products or from degradation. Possibly two of these substances differ only in an isoaspartyl instead of an aspartyl binding. A baseline separation was possible with an acetate buffer (pH 4.4, 60 mM) containing 0.3% (m/m) PEG 20,000 and 0.1 mM Zn2+. The possibilities to prevent wall adsorption are discussed.
Dette
C
C
Institute of Pharmacy and Food Chemistry, University of Würzburg, Germany.
Wätzig
H
H
D000085:Acetates; D000595:Amino Acid Sequence; D002021:Buffers; D002197:Capillary Action; D004586:Electrophoresis / Q000379:methods*; D006629:Hirudins / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D011092:Polyethylene Glycols; D013917:Thrombin / Q000037:antagonists & inhibitors*; D015032:Zinc
D000085:Acetates; D002021:Buffers; D006629:Hirudins; D011092:Polyethylene Glycols; D013917:Thrombin; D015032:Zinc