[Analysis of recombinant erythropoietin and its tryptic digest by capillary electrophoresis and capillary electrophoresis-electrospray ionization-mass spectrometry using capillary coated with 6,6-ionene].
The microheterogeneity of recombinant human erythropoietin (rhEPO) was analyzed by capillary electrophoresis (CE) using a capillary coated with 6,6-ionene. The applicability of a volatile electrolyte for fast analysis of tryptic fragments of rhEPO with online CE-electrospray ionization-mass spectrometry (ESI-MS) was investigated, resulting in a reproducible separation of eleven rhEPO tryptic fragments within 22 min under the following conditions: running buffer 300 mmol/L acetic acid-ammonium acetate (HAc-NH4Ac), pH 4.80, separation voltage -15 kV and capillary temperature 25 degrees C. The proposed method is rapid and effective, and can be used for the structural analysis of related proteins.
Yu
Bing
B
The Key Lab of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing, China. yubing@pku.edu.cn
Cong
Hailin
H
Liu
Huwei
H
Li
Yuanzong
Y
Liu
Feng
F
D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods*; D004921:Erythropoietin / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D011108:Polymers / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D011994:Recombinant Proteins; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation* / Q000379:methods*; D014357:Trypsin / Q000378:metabolism*
C511857:6,6-ionene; D011108:Polymers; D000644:Quaternary Ammonium Compounds; D011994:Recombinant Proteins; D004921:Erythropoietin; D014357:Trypsin