Carrier ampholyte‐free free‐flow isoelectric focusing for separation of protein

2019 ◽  
Author(s):  
Shuai Wang ◽  
Lingyi Zhang ◽  
Haofan Sun ◽  
Zhanying Chu ◽  
Haihong Chen ◽  
...  
Keyword(s):  
Isoelectric Focusing ◽  
Free Flow ◽  
Carrier Ampholyte

Free-flow electrophoresis for the purification of proteins: II. Isoelectric focusing and field step electrophoresis

1990 ◽  
Vol 11 (11) ◽  
pp. 942-947 ◽  
Author(s):  
Reinhard Kuhn ◽  
Sabrina Hoffstetter-Kuhn ◽  
Horst Wagner
Keyword(s):  
Isoelectric Focusing ◽  
Free Flow

Native protein blotting after isoelectric focusing in fabric reinforced polyacrylamide gels in carrier ampholyte generated or immobilized pH gradients

1988 ◽  
Vol 9 (9) ◽  
pp. 497-511 ◽  
Author(s):  
Angelika Kinzkofer-Peresch ◽  
Nikos P. Patestos ◽  
Markus Fauth ◽  
Friedrich Kögel ◽  
Roland Zok ◽  
...  
Keyword(s):  
Isoelectric Focusing ◽  
Polyacrylamide Gels ◽  
Native Protein ◽  
Ph Gradients ◽  
Protein Blotting ◽  
Carrier Ampholyte

Free Flow Isoelectric Focusing in a Microfluidic Device

2014 ◽  
Author(s):  
Kisoo Yoo ◽  
Prashanta Dutta ◽  
Jin Liu
Keyword(s):  
Mathematical Model ◽  
Isoelectric Focusing ◽  
Microfluidic Device ◽  
Microfluidic Chip ◽  
Cardiac Troponin ◽  
Troponin I ◽  
Free Flow ◽  
Charged Species ◽  
Simulation Results ◽  
Separation Of Proteins

In recent years, there are growing interests in the use of free flow isoelectric focusing (FFIEF). In FFIEF, a thin sheath of laminar flow is introduced perpendicular to the direction of the applied electric field for continuous separation of proteins and charged species. This technique is especially useful in microfluidic device since the electrophoretically separated bands do not have to be mobilized for detection or further analysis. In this study, a mathematical model is developed to simulate free flow isoelectric process in microfluidic devices considering electroneutrality and incompressibility of electrolytes. Our mathematical model is based on mass, momentum and charge conservation equations. A finite volume based numerical scheme is implemented to simulate two dimensional FFIEF in a microfluidic chip. Simulation results indicate that pH gradient forms as samples flow downstream and proteins can be separated effectively using this technique. A new design of microfluidic chip is proposed for separation for cardiac troponin I from serum albumin using FFIEF technique.

Analytical Methods and Bench Scale Preparative Bioseparations

2015 ◽  
Author(s):  
Roger G. Harrison ◽  
Paul W. Todd ◽  
Scott R. Rudge ◽  
Demetri P. Petrides
Keyword(s):  
Isoelectric Focusing ◽  
Analytical Methods ◽  
Process Development ◽  
Scale Up ◽  
Free Flow ◽  
Analytical Methodology ◽  
Bench Scale ◽  
Magnetic Separations ◽  
Analytical Development ◽  
Downstream Processes

The development of efficient and reliable processes for bioseparations is dependent on the availability of suitable analytical methods. This means it is important that work on analytical methodology for the bioproduct of interest starts at the very beginning of process development. Analytical studies are important throughout the development and scale up of the process, as changes can occur either to the product or to its associated impurities from what may be thought of as minor changes in the process. This chapter gives access to the vocabulary and techniques used in quality control and analytical development activities, starting with a description of specifications typically set for a pharmaceutical and the rationale behind them. Then, before discussing the assays themselves, we describe assay attributes, which can be measured and used to help not only the assay developer but also the biochemist and engineer responsible for developing downstream processes determine the usefulness and meaning of the assay. Finally, we turn to assays that are commonly applied in biotechnology, as they apply to biological activity, identity, and purity. These assays are the ultimate yardsticks by which the process is measured. Purification methods are developed for their ability to remove a contaminant from the product of interest, whether it is a related molecule, a contaminant related to a host organism, such as DNA or endotoxin, or a process contaminant, such as a residual solvent or water. Critical to understanding process performance is an understanding of how the assays that measure these contaminants have been developed, what the assay strengths and limitations are, and what they indicate and why. Electrophoresis and magnetic separation are two methods that are now used for the bench scale preparative purification of bioproducts, including living cells. The electrophoresis systems with the highest capacity are free-flow electrophoresis, density gradient electrophoresis, recycling free-flow isoelectric focusing, and rotating isoelectric focusing, and the principles of operation of these are discussed. The physical principles of magnetic separations are presented, as well as magnetic reagents and applications of magnetic separators.

Free-flow isoelectric focusing of proteins remaining in cell fragments following sonication of thyroid carcinoma cells

2005 ◽  
Vol 26 (11) ◽  
pp. 2109-2116 ◽  
Author(s):  
Christian Obermaier ◽  
Vera Jankowski ◽  
Cornelia Schmutzler ◽  
Johann Bauer ◽  
Robert Wildgruber ◽  
...  
Keyword(s):  
Thyroid Carcinoma ◽  
Isoelectric Focusing ◽  
Carcinoma Cells ◽  
Free Flow ◽  
Cell Fragments
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