title: Characterization of human growth hormone by capillary electrophoresis.
abstract: Production of proteins by recombinant DNA technology for use as pharmaceuticals requires the use of the most powerful tools of analytical protein chemistry in order to confirm purity and identity of the product and reliability of the process. Capillary electrophoresis is an emerging technology that shows high sensitivity and selectivity and may have promise in this application. The technique combines the instrumental control and quantification features of high-performance liquid chromatography with the separating power of electrophoresis, and thereby has attracted broad interest. In this report, human growth hormone expressed in bacteria has been analyzed by both free zone electrophoresis and isoelectric focusing in a coated capillary to demonstrate the separation of the native molecule from its deamidated variant. A capillary zone electrophoretic tryptic map has also been developed and characterized. This map complements the widely employed reversed-phase high-performance liquid chromatography tryptic mapping systems that are important in protein characterization. Certain drawbacks to capillary zone electrophoresis compared to other analytical methods are noted, including relatively poor reproducibility and low sample tolerance. For applications as demonstrated here, however, the speed, separating power and sensitivity of the technique compensate for these shortcomings.
authors.affiliation: Department of Medicinal and Analytical Chemistry, Genentech, Inc., South San Francisco, CA 94080.
authors.forename: J
authors.initials: J
authors.lastname: Frenz
authors.forename: S L
authors.initials: SL
authors.lastname: Wu
authors.forename: W S
authors.initials: WS
authors.lastname: Hancock
mesh_terms: D002852:Chromatography, Ion Exchange; D004586:Electrophoresis / Q000379:methods*; D013006:Growth Hormone / Q000032:analysis*; D006801:Humans; D007525:Isoelectric Focusing
chemical_list: D013006:Growth Hormone