Capillary Electrophoretic Separation of Human Recombinant Erythropoietin (r-HuEPO) Glycoforms

✍ Scribed by E. Watson; F. Yao

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 336 KB
• Volume: 210
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1212

✦ Synopsis

Free zone capillary electrophoresis was investigated for the separation of glycoforms in recombinant erythropoietin ( (r)-HuEPO). Reduction of electroendoosmotic flow through the addition of 1,4 -diaminobutane and in the presence of urea resulted in optimum separation of all of the major glycoforms. Incubation of r-HuEPO with neuraminidase caused the slowest migrating species to diminish first in response, indicating that these contained the highest numbers of sialic acids. Additional evidence was obtained from spiking r-HuEPO with individual glycoforms isolated by preparative isoelectric focusing. These results indicate that the separation occurs in a predictable manner in order of increasing number of sialic acids contained in the glycoforms. As a consequence, the electropherograms provide the same information as that obtained from conventional gel isoelectric focusing, although separating by a different mechanism. 1893 Academic Press, Inc.