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Related Concept Videos

Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
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The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
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Proteins show rotational as well as lateral diffusion across the membrane. The lateral diffusion of proteins was confirmed through the cell fusion experiment where mouse and human cells were fused, resulting in hybrid cells. When the human and mouse cells fused, the specific membrane proteins on human and mouse cells were marked with the red and green-fluorescent markers, respectively. Initially, the red and green fluorescence was located on the respective hemisphere of the cell. As time...
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Diffusion Split-Flow Thin Cell (SPLITT) system for protein separations.

Srinivas Merugu1, Himanshu J Sant, Bruce K Gale

  • 1State of Utah Center of Excellence for Biomedical Microfluidics, Department of Electrical Engineering, University of Utah, 50 S. Central Campus Drive, Rm. 2110, Salt Lake City, UT 84112, USA. merugu s@yahoo.com

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|July 17, 2012
PubMed
Summary
This summary is machine-generated.

A novel diffusion Split-Flow Thin Cell (SPLITT) system effectively removes small peptides like β2-microglobulin (β2M) and parathyroid hormone (PTH) from blood, preserving larger proteins. This technology shows promise for enhancing kidney dialysis by removing toxins.

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Area of Science:

  • Biomolecular Engineering
  • Separation Science
  • Biotechnology

Background:

  • Current kidney dialysis methods struggle to remove smaller protein-based toxins.
  • There is a need for advanced separation techniques to selectively remove specific molecules from biological fluids.

Purpose of the Study:

  • To develop and validate a diffusion-based Split-Flow Thin Cell (SPLITT) system for selective peptide removal.
  • To optimize operating conditions using numerical modeling for efficient protein purification.

Main Methods:

  • Fabrication of a diffusion SPLITT system using xurography.
  • Development of a 2D numerical model based on Navier-Stokes and convection-diffusion equations.
  • Experimental validation of the SPLITT system for removing β2-microglobulin (β2M) and parathyroid hormone (PTH) while retaining albumin.

Main Results:

  • The diffusion SPLITT system successfully removed over 25% of small peptides (β2M, PTH) while preserving over 97% of larger proteins like albumin.
  • Experimental results showed good agreement with predictions from the numerical model.
  • The system demonstrated continuous purification based on molecular size and diffusion coefficients.

Conclusions:

  • The diffusion SPLITT system offers a promising continuous method for selective removal of small peptides from biological samples.
  • This technique has potential applications in improving kidney dialysis by removing toxins not cleared by conventional methods.
  • The developed numerical model aids in optimizing SPLITT system performance for various purification tasks.