Online transient isotachophoresis concentration by the pseudo-terminating electrolyte buffer for the separation of DNA-aptamer and its thrombin complex in poly(methyl methacrylate) microchip

Jun Wang, Yong Zhang, Yukihiro Okamoto, Noritada Kaji, Manabu Tokeshi, Yoshinobu Baba

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Online automatic transient isotachophoresis concentration of DNA-aptamer and its thrombin complex by using one kind of pseudo-terminating electrolyte buffer in a cross-channel poly(methyl methacrylate) microchip is reported. Sample injection, transient concentration and separation were done continuously and controlled by a sequential voltage switching program, time-consuming steps and complicated chip design were not required. Peak resolution between DNA-aptamer and its thrombin complex was influenced by this novel pseudo-terminating electrolyte buffer, which was prepared by the addition of chemical component with slow mobility into the same buffer as leading electrolyte buffer. 1100-fold signal enhancement of thrombin complex was achieved by this transient isotachophoresis on a standard cross-form microchip. The concentration effect or standing time of transient isotachophoresis was proved to be influenced by the concentration of leading electrolyte ion and the concentration of pseudo-terminating electrolyte buffer ion (glycine). The transient concentration was followed by on-chip nondenaturing gel electrophoresis in methylcellulose solution for the size-based separation. The detection limit, taken as the lowest thrombin concentration at threefold S/N, was determined to be 0.5 amol in mass by this method.

Original languageEnglish
Pages (from-to)1142-1147
Number of pages6
JournalAnalyst
Volume136
Issue number6
DOIs
Publication statusPublished - Mar 21 2011
Externally publishedYes

Fingerprint

Isotachophoresis
Nucleotide Aptamers
Polymethyl Methacrylate
Polymethyl methacrylates
Thrombin
electrolyte
Electrolytes
Buffers
DNA
Ions
Methylcellulose
ion
Electrophoresis
Glycine
Limit of Detection
Amino acids
electrokinesis
Gels
gel
fold

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

Online transient isotachophoresis concentration by the pseudo-terminating electrolyte buffer for the separation of DNA-aptamer and its thrombin complex in poly(methyl methacrylate) microchip. / Wang, Jun; Zhang, Yong; Okamoto, Yukihiro; Kaji, Noritada; Tokeshi, Manabu; Baba, Yoshinobu.

In: Analyst, Vol. 136, No. 6, 21.03.2011, p. 1142-1147.

Research output: Contribution to journalArticle

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