Simultaneous determination of 19 intracellular nucleotides and nucleotide sugars in Chinese Hamster ovary cells by capillary electrophoresis.
Twelve nucleotides and seven nucleotide sugars in Chinese Hamster ovary (CHO) cells were determined by capillary electrophoresis (CE). The CE operating conditions of buffer pH value, ion strength, capillary temperature, polymer additive and cell extraction method were investigated. Optimum separation was achieved with 40 mM sodium tetraborate buffer (pH 9.5) containing 1% (w/v) polyethylene glycol (PEG) at a capillary temperature of 22 degrees C. Acetonitrile and chloroform were used for intracellular extraction. This method can be used to monitor intracellular carbohydrate metabolism.
Feng
Hua-Tao
HT
Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore. feng_huatao@bti.a-star.edu.sg
Wong
Niki
N
Wee
Sheena
S
Lee
May May
MM
D000818:Animals; D016466:CHO Cells; D002457:Cell Extracts / Q000032:analysis; D006224:Cricetinae; D003412:Cricetulus; D019075:Electrophoresis, Capillary / Q000379:methods*; D009702:Nucleoside Diphosphate Sugars / Q000032:analysis*; D009711:Nucleotides / Q000032:analysis*
D002457:Cell Extracts; D009702:Nucleoside Diphosphate Sugars; D009711:Nucleotides