Characterization and Optimization of Isotacophoersis Parameters for Pacific Blue Succinimidyl Ester Dye on a PDMS Microfluidic Chip
| Autor: | Jungtae Kim, Zachary Estlack, Jungkyu Kim, Jasmine Pramila Devadhasan, Himali Somaweera |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Resolution (mass spectrometry)
Hydrochloride lcsh:Mechanical engineering and machinery isotachophoresis pacific blue succinimidyl ester 02 engineering and technology Electrolyte 01 natural sciences Article chemistry.chemical_compound PDMS (polydimethylsiloxane) microfluidic chip Electric field preconcentration lcsh:TJ1-1570 Electrical and Electronic Engineering Chromatography Quenching (fluorescence) Polydimethylsiloxane Chemistry Mechanical Engineering 010401 analytical chemistry lactate dehydrogenase 021001 nanoscience & nanotechnology 0104 chemical sciences Microfluidic chip Control and Systems Engineering Isotachophoresis 0210 nano-technology |
| Zdroj: | Micromachines Volume 11 Issue 11 Micromachines, Vol 11, Iss 951, p 951 (2020) |
| ISSN: | 2072-666X |
| DOI: | 10.3390/mi11110951 |
| Popis: | Isotachophoresis (ITP) for Pacific Blue (PB) dye using a polydimethylsiloxane (PDMS) microfluidic chip is developed and characterized by determining the types and concentrations of electrolytes, the ITP duration, and the electric field density. Among candidate buffers for the trailing electrolyte (TE) and leading electrolyte (LE), 40 mM borate buffer (pH 9) and 200 mM trisaminomethane hydrochloride (Tris-HCl) (pH 8) were selected to obtain the maximum preconcentration and resolution of the PB bands, respectively. With the selected TE and LE buffers, further optimization was performed to determine the electric field (EF) density and the ITP duration. These ITP parameters showed a 20&ndash 170,000 preconcentration ratio from initial PB concentrations of 10 nM&ndash 100 fM. Further demonstration was implemented to preconcentrate PB-conjugated lactate dehydrogenase (LDH) using the PDMS microfluidic chip. By utilizing the quenching nature of PB-LDH conjugation, we were able to identify concentrations of LDH as low as 10 ng/mL. This simple PDMS microfluidic chip-based ITP for PB preconcentration enables highly sensitive biological and chemical analyses by coupling with various downstream detection systems. |
| Databáze: | OpenAIRE |
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