Exceeding 20,000-fold concentration of protein by the on-line isotachophoresis concentration in poly(methyl methacrylate) microchip.
In this research, a simple on-line microchip gel electrophoresis with ITP was applied for the concentration and separation of BSA and its immunoassay complex with mAb in a single cross form PMMA microchip. We investigated the ITP concentration effect in PMMA MCE using combination of leading electrolytes, terminating electrolytes and other factors. We realized an ITP-based concentration and separation of BSA and its immunoassay complexes in standard cross-channel microchip gel electrophoresis, which exceeded 2000-fold concentration of BSA immunocomplex using Tris-H3PO4 as a leading electrolyte and Tris-gamma-amino butyric acid as a terminating electrolyte. In addition, we also realized concentration of BSA sample in water, which was more than 20 000-fold and was the result of the concentration effect from combining ITP and the sample stacking techniques.
Wang
Jun
J
Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Nagoya, Japan. jwang@pme.coe.nagoya-u.ac.jp
Zhang
Yong
Y
Mohamadi
Mohamad Reza
MR
Kaji
Noritada
N
Tokeshi
Manabu
M
Baba
Yoshinobu
Y
D000911:Antibodies, Monoclonal / Q000378:metabolism; D004573:Electrolytes / Q000737:chemistry; D004586:Electrophoresis / Q000379:methods*; D007118:Immunoassay / Q000379:methods*; D008747:Methylcellulose / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D005680:gamma-Aminobutyric Acid / Q000737:chemistry
D000911:Antibodies, Monoclonal; D004573:Electrolytes; D012710:Serum Albumin, Bovine; D005680:gamma-Aminobutyric Acid; D008747:Methylcellulose; D019904:Polymethyl Methacrylate