Continuous on-chip fluorescence labelling, free-flow isoelectric focusing and marker-free isoelectric point determination of proteins and peptides
نویسندگان
چکیده
منابع مشابه
Gradient-free determination of isoelectric points of proteins on chip.
The isoelectric point (pI) of a protein is a key characteristic that influences its overall electrostatic behaviour. The majority of conventional methods for the determination of the isoelectric point of a molecule rely on the use of spatial gradients in pH, although significant practical challenges are associated with such techniques, notably the difficulty in generating a stable and well cont...
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The separation of ampholytic components according to isoelectric point has played an important role in isolating, reducing complexity and improving peptide and protein detection. This brief review outlines the basics of isoelectric focusing, including a summary of the historical achievements and considerations in experimental design. Derivative methodologies of isoelectric focusing are also dis...
متن کاملLabel-free microfluidic free-flow isoelectric focusing, pH gradient sensing and near real-time isoelectric point determination of biomolecules and blood plasma fractions.
We demonstrate the fabrication, characterization and application of microfluidic chips capable of continuous electrophoretic separation via free flow isoelectric focussing (FFIEF). By integration of a near-infrared (NIR) fluorescent pH sensor layer under the whole separation bed, on-line observation of the pH gradient and determination of biomolecular isoelectric points (pI) was achieved within...
متن کاملMicrofluidic Protein Separation by Free Flow Isoelectric Focusing
Disposable, inexpensive microfluidic devices have the potential to become a robust tool for proteomic research for separating and identifying proteins and protein complexes. In this work, a preparative scale free flow IEF isoelectric focusing (FFIEF) device was designed, investigated, and optimized. A constant electric field of up to 350V/cm was generated across the separation channel upon opti...
متن کاملOn-chip coupling of isoelectric focusing and free solution electrophoresis for multidimensional separations.
We have developed an acrylic microfluidic device that sequentially couples liquid-phase isoelectric focusing (IEF) and free solution capillary electrophoresis (CE). Rapid separation (<1 min) and preconcentration (73x) of species were achieved in the initial IEF dimension. Using full-field fluorescence imaging, we observed nondispersive mobilization velocities on the order of 20 microm/s during ...
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ژورنال
عنوان ژورنال: Lab on a Chip
سال: 2016
ISSN: 1473-0197,1473-0189
DOI: 10.1039/c6lc00055j