Rapid and subnanomolar assay of recombinant human erythropoietin by capillary electrophoresis using NanoOrange precolumn labeling and laser-induced fluorescence detection

doi:10.1002/jssc.201400263

	Rapid and subnanomolar assay of recombinant human erythropoietin by capillary electrophoresis using NanoOrange precolumn labeling and laser-induced fluorescence detection
	Pang, Nannan 1,2; Bai, Yu 2; Zhou, Yu 2; Yang, Xia 2; Zhang, Zhengxiang 2; Nie, Honggang 2; Fu, Xiaofang 2; Liu, Huwei 2
刊名	JOURNAL OF SEPARATION SCIENCE
	2014-08-01
卷号	37 期号:16 页码:2233-2238
关键词	Capillary Electrophoresis Laser-induced Fluorescence Nanoorange Recombinant Human Erythropoietin
ISSN号	1615-9306
DOI	10.1002/jssc.201400263
文献子类	Article
英文摘要	Because of less functionally critical carbohydrate sectors that contributed to the stability, efforts have been made to quantify intact recombinant human erythropoietin. A simple, rapid capillary electrophoresis with laser-induced fluorescence method for the assay of recombinant human erythropoietin was developed, with a limit of detection of intact recombinant human erythropoietin at subnanomolar concentration (up to 10 ng/mL or 3 x 10(-10) M), which is among the lowest reported. High sensitivity was accomplished by precolumn derivatization with the noncovalent dye NanoOrange. Capillary electrophoresis separation and reaction conditions were carefully manipulated for avoiding microheterogeneity of glycoforms and inhomogeneity of multiple labeling products. The fluorescence signal was linear over the range of 10 ng/mL-10 mu g/mL, corresponding to the detection requirement of recombinant human erythropoietin in biofluids and pharmaceutical samples, as demonstrated by a real sample analysis. Although the salt in reaction mixtures showed a detrimental effect on the fluorescence of the derivatives, this method could tolerate a certain amount of salt, extending its application in biofluid analysis. In addition, zero-order fluorescence emission kinetics was obtained, indicating that the rapid decay of recombinant human erythropoietin was derived from a self-quenching effect.
WOS关键词	IONIZATION MASS-SPECTROMETRY ; COLUMN IMAGING DETECTION ; EQUINE PLASMA ; STIMULATING PROTEIN ; DARBEPOETIN-ALPHA ; COATED CAPILLARY ; DOPING CONTROL ; LC-MS/MS ; GLYCOFORMS ; DIFFERENTIATION
WOS研究方向	Chemistry
语种	英语
出版者	WILEY-V C H VERLAG GMBH
WOS记录号	WOS:000340556300019
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/167300]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Liu, Huwei
作者单位	1.Univ Sci & Technol, Sch Chem & Biol Engn, Beijing, Peoples R China 2.Peking Univ, Beijing Natl Lab Mol Sci, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Inst Analyt Chem,Coll Chem & Mol Engn, Beijing 100871, Peoples R China
推荐引用方式 GB/T 7714	Pang, Nannan,Bai, Yu,Zhou, Yu,et al. Rapid and subnanomolar assay of recombinant human erythropoietin by capillary electrophoresis using NanoOrange precolumn labeling and laser-induced fluorescence detection[J]. JOURNAL OF SEPARATION SCIENCE,2014,37(16):2233-2238.
APA	Pang, Nannan.,Bai, Yu.,Zhou, Yu.,Yang, Xia.,Zhang, Zhengxiang.,...&Liu, Huwei.(2014).Rapid and subnanomolar assay of recombinant human erythropoietin by capillary electrophoresis using NanoOrange precolumn labeling and laser-induced fluorescence detection.JOURNAL OF SEPARATION SCIENCE,37(16),2233-2238.
MLA	Pang, Nannan,et al."Rapid and subnanomolar assay of recombinant human erythropoietin by capillary electrophoresis using NanoOrange precolumn labeling and laser-induced fluorescence detection".JOURNAL OF SEPARATION SCIENCE 37.16(2014):2233-2238.