Rapid and subnanomolar assay of recombinant human erythropoietin by capillary electrophoresis using NanoOrange precolumn labeling and laser-induced fluorescence detection
文献类型：期刊论文
| 作者 | Pang, Nannan1,2; Bai, Yu2; Zhou, Yu2; Yang, Xia2; Zhang, Zhengxiang2; Nie, Honggang2; Fu, Xiaofang2; Liu, Huwei2 |
| 刊名 | JOURNAL OF SEPARATION SCIENCE
 |
| 出版日期 | 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 |
| WOS记录号 | WOS:000340556300019 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| 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. |
入库方式： OAI收割
来源：大连化学物理研究所
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