iBodies against Different Protease Families

We are currently working on development of iBodies® directed towards whole protease families

PROTEASES are enzymes that accelerate the hydrolysis of peptide bonds. iBodies® accessorized with inhibitors specific to whole families of these enzymes are under development. Currently, two of the main protease groups; aspartic and serine proteases are being explored.

ASPARTIC PROTEASE FAMILY

Targeted Biomolecule Specification:

A family of enzymes that cleaves peptide bonds in a single step proteolysis via utilization of an activated water molecule by one or more aspartate residues. Almost all aspartic proteases are inhibited by pepstatin. Pepsins, cathepsins and renins are representatives of the eukaryotic aspartic proteases.

Targeting ligand:

Pepstatin based inhibitor

Binding kinetics parameters:

iBody KD = Ki (HIV-1 protease) = 19 n (measured with Surface Plasmon Resonance)

Experiments and potential uses:

anti-aspartic protease family iBodies® were used in methods such as bio-imaging with confocal microscopy, ELISA, protein immobilization and pull down from cell lysates (see the results below)

References:

Tang, J. and Wong, R. N. S. (1987), Evolution in the structure and function of aspartic proteases. J. Cell. Biochem., 33: 53–63. doi:10.1002/jcb.240330106

 

SERINE PROTEASE FAMILY

Targeted Biomolecule Specification:

Serine proteases (SerPR) are peptidases with a catalytic triad (His, Ser, Asp) in their active site. Two broad groups of these enzymes are the chymotrypsin-like and subtilisin-like proteases.

Targeting ligand:

Pefabloc (4-(2-aminoethyl) benzenesulfonyl fluoride) based inhibitor

Experiments and potential uses:

anti-serine protease family iBodies® were used in methods such as bio-imaging with confocal microscopy, ELISA, protein immobilization, pull down from cell lysates and western blotting (see the results below)

 

References:

Di Cera, E. (2009). Serine Proteases. IUBMB Life, 61(5), 510–515. http://doi.org/10.1002/iub.186