title: Analysis of a biopolymer by capillary electrophoresis with a chemiluminescence detector using a polymer solution as the separation medium.
abstract: We developed capillary electrophoresis with a chemiluminescence detector using a polymer solution as the separation medium for the analysis of biopolymers, such as DNA and protein. A peroxyoxalate chemiluminescence reagent of bis(2,4,6-trichlorophenyl)oxalate was used together with fluorescein-labeling reagent. When a migration buffer solution containing carboxylmethylcellulose was used, the flow-type chemiluminescence detection cell was found to give a better resolution than the batch-type one. Fluorescein-labeled adenosine triphosphate of 1.0 x 10(-4) M was examined by means of capillary electrophoresis with absorption (260 nm), fluorescence (ex. 496 nm and em. 517 nm), and chemiluminescence detectors. The chemiluminescence detection showed the highest sensitivity among them; the S/N ratios obtained by absorption, fluorescence, and chemiluminescence detections were 4, 38, and 130, respectively. Fluorescein-labeled DNA was prepared through a polymerase chain reaction using fluorescein-labeled deoxyadenosine triphosphate. A mixture of the labeled DNA fragments (500, 600, 700, 800, 900, and 993 bp) was successfully separated and detected by the present system. A mixture of proteins (lysozyme, cytochrome C, and ribonuclease A) which were labeled with fluorescein isothiocyanate was also separated and detected.
authors.affiliation: Department of Chemical Engineering and Materials Science, Faculty of Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Tsukagoshi
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Shikata
authors.forename: Riichiro
authors.initials: R
authors.lastname: Nakajima
authors.forename: Masaharu
authors.initials: M
authors.lastname: Murata
authors.forename: Mizuo
authors.initials: M
authors.lastname: Maeda
mesh_terms: D000042:Absorption; D000255:Adenosine Triphosphate / Q000032:analysis; D020029:Base Pairing; D001704:Biopolymers / Q000032:analysis* / Q000737:chemistry*; D002021:Buffers; D003574:Cytochrome c Group / Q000032:analysis / Q000737:chemistry; D004247:DNA / Q000032:analysis / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D019793:Fluorescein; D008163:Luminescent Measurements; D009113:Muramidase / Q000032:analysis / Q000737:chemistry; D010070:Oxalates; D011108:Polymers / Q000737:chemistry*; D015203:Reproducibility of Results; D012259:Ribonuclease, Pancreatic / Q000032:analysis / Q000737:chemistry; D012680:Sensitivity and Specificity; D012996:Solutions / Q000737:chemistry; D013997:Time Factors
chemical_list: D001704:Biopolymers; D002021:Buffers; D003574:Cytochrome c Group; D010070:Oxalates; D011108:Polymers; D012996:Solutions; C036521:bis(2,4,6-trichlorophenyl)oxalate; D000255:Adenosine Triphosphate; D004247:DNA; D012259:Ribonuclease, Pancreatic; D009113:Muramidase; D019793:Fluorescein