EXAMPLE: PROTEASES

Proteases and their regulation by specific inhibitors play besides their function in numerous metabolic pathways a central role in the progression of pathogenic forms of apotosis, in tumour growth and other diseases like Alzheimer or HIV.
BIA technology provides detailed insight in regulatory mechanisms of inhibitor binding to the protease. Different Analysis strategies offer kinetic views of the interactions by alternative experimental approaches. It is possible to immobilise either the inhibitors (e.g. peptides) on the chip surface to monitor directly the binding of the proteases or to couple the proteases to detect the binding of the low molecular weight inhibitors.

 

For kinetic analysis of the interaction of low molecular inhibitors competition assays and the determination of EC50-values are most suitable.

Biaffin has developed coupling and analysis procedures for reproducible conditions (even at low pH) for the interaction analysis of proteases. These assays can readily be transferred to other proteases.

 

On request we can provide you with detailed information about suitable protease assays designed for your individual problem.

different concentrations of protease cathepsin D binding to a coupled inhibitor

The figure above shows the binding of protease cathepsin D to a coupled inhibitor.

solution competition assay of cathepsin D with the protease inhibitor pepstatin

The solution competition assay of this interaction with the protease inhibitor pepstatin reveals an EC50 = 0.1 nM.

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Tel.: +49 (0) 561-804 4661 | Fax: +49 (0) 561-804 4665 | info@biaffin.de