Ionene-dynamically coated capillary for analysis of urinary and   recombinant human erythropoietin by capillary electrophoresis and online   electrospray ionization mass spectrometry

doi:10.1002/jssc.200500156

CORC > 北京大学 > 化学与分子工程学院

	Ionene-dynamically coated capillary for analysis of urinary and recombinant human erythropoietin by capillary electrophoresis and online electrospray ionization mass spectrometry
	Yu, B ; Cong, HL ; Liu, HW ; Li, YZ ; Liu, F
刊名	journal of separation science
	2005
关键词	coating erythropoietin glycoforms ionene polymer online-capillary electrophoresis-electrospray ionization mass spectrometry (CE-ESI-MS) ASSISTED-LASER-DESORPTION/IONIZATION ZONE-ELECTROPHORESIS LIQUID-CHROMATOGRAPHY PROTEINS COATINGS GLYCOPROTEINS GLYCOFORMS SEPARATION FRAGMENTATION GLYCOSYLATION
DOI	10.1002/jssc.200500156
英文摘要	In this article, a series of ionene polymers were synthesized and used to coat fused-silica capillaries for the separation of recombinant and urinary human erythropoietin (rhEPO and uEPO) standards by CE. The influence of the charge density of coatings on the separation of rhEPO and uEPO glycoforms was investigated. Then, we further studied the method for fast separation and detection of rhEPO and uEPO standards by CE-ESI-MS. The influence of several CE and MS operating parameters, such as the concentration of CE running buffer, applied external pressure, and the composition and flow rate of sheath liquid on CE-ESI-MS was studied. The results demonstrated that when the capillary was permanently coated with 6,6-ionene and the pH value of acetic acid-ammonium acetate running buffer was 4.80 and 5.50, respectively, a significantly reproducible separation was achieved for rhEPO and uEPO glycoforms. In the online CE-ESI-MS experiments, we not only achieved the online MS signal of uEPO, but also obtained baseline separation of three major rhEPO glycoforms successfully and reproducibly on the 6,6-ionene-coated capillaries. Furthermore, the standard mixture of rhEPO and uEPO was separated, and two incompletely resolved peaks that were identified to be rhEPO and uEPO by the unique MS "fingerprint" were obtained. Additionally, the molecular weight of rhEPO and uEPO were verified and compared to the results by MALDI-TOF-MS. It can be concluded that, in contrast to other indirect methods, the online CE-ESI-MS technique with the combination of the advantages of both CE and MS shows great potential for the separation and detection of rhEPO doping directly in competitive sports.; Chemistry, Analytical; SCI(E); EI; PubMed; 28; ARTICLE; 17; 2390-2400; 28
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/199433]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Yu, B,Cong, HL,Liu, HW,et al. Ionene-dynamically coated capillary for analysis of urinary and recombinant human erythropoietin by capillary electrophoresis and online electrospray ionization mass spectrometry[J]. journal of separation science,2005.
APA	Yu, B,Cong, HL,Liu, HW,Li, YZ,&Liu, F.(2005).Ionene-dynamically coated capillary for analysis of urinary and recombinant human erythropoietin by capillary electrophoresis and online electrospray ionization mass spectrometry.journal of separation science.
MLA	Yu, B,et al."Ionene-dynamically coated capillary for analysis of urinary and recombinant human erythropoietin by capillary electrophoresis and online electrospray ionization mass spectrometry".journal of separation science (2005).