A new impedance based approach to test the activity of recombinant protein – Semaphorins as a test case

The biological activity of a recombinant protein is routinely measured using a bioassay such as an enzyme assay. However, many proteins have no enzymatic activity and in many cases it is difficult to devise a simple and reliable approach to test their activity. Semaphorins, Ephrins, Slits, Netrins o...

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Bibliographic Details
Published in:European journal of cell biology 2015-10, Vol.94 (10), p.453-457
Main Authors: Birger, Anastasya, Besser, Elazar, Reubinoff, Benjamin, Behar, Oded
Format: Article
Language:English
Subjects:
Animals
Biological Assay - methods
Cytoskeleton
Electric Impedance
Humans
Ligands
Nerve Tissue Proteins - analysis
Nerve Tissue Proteins - genetics
Nerve Tissue Proteins - metabolism
Receptors, Cell Surface - analysis
Receptors, Cell Surface - genetics
Receptors, Cell Surface - metabolism
Recombinant Proteins - analysis
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Semaphorins - analysis
Semaphorins - genetics
Semaphorins - metabolism
Transfection
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Summary:The biological activity of a recombinant protein is routinely measured using a bioassay such as an enzyme assay. However, many proteins have no enzymatic activity and in many cases it is difficult to devise a simple and reliable approach to test their activity. Semaphorins, Ephrins, Slits, Netrins or amylin-assisted proteins have numerous activities affecting many systems and cell types in the human body. Most of them are also able to induce rapid cytoskeleton changes at least in some cell types. We assumed therefore, that such proteins might be tested based on their ability to modulate the cytoskeleton. Here we tested a number of semaphorins in an impedance based label-free platform that allows for dynamic monitoring of subtle morphological and adhesive changes. This system has proved to be a very fast, sensitive and effective way to monitor and determine the activity of such proteins. Furthermore we showed that it is possible to customize a cell-protein system by transfecting the cells with specific receptors and test the cell response following the addition of the recombinant ligand protein. Since other protein families such as Ephrins and Netrins can also influence the cytoskeleton of some cells, this approach may be applicable to a large number of proteins.
ISSN:0171-9335
1618-1298
DOI:10.1016/j.ejcb.2015.06.006