Modification of enzymatic crosslinkers for controlling properties of crosslinked matrices
AssigneeLifebond Ltd.
InventorsOrahn Preiss-Bloom, Guy Tomer
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Abstract
Improved matrix or hydrogel that is formed by enzymatic crosslinking of polymers wherein the crosslinking enzyme molecules have been modified for the purpose of improving the crosslinking density, mechanical properties, or other properties of the matrix, and/or to provide improved control over the rate and/or extent of crosslinking, wherein the enzyme molecules are modified to alter the perceived volume of the enzyme molecules in the crosslinked matrix being formed. Methods of production and of use are also provided.
Claims54
A cross-linked matrix, comprising a substrate polymer and a modified enzyme molecule, wherein said substrate polymer has been crosslinked by said modified enzyme molecule, wherein both said modified enzyme molecule and said substrate polymer are present in the matrix, said modified enzyme molecule having a modification selected from the group consisting of:
- a modification that increases the actual size of said enzyme molecule;
- or
- a modification that increases the hydrodynamic volume of said enzyme molecule
- wherein said modified enzyme has a reduced cross-linking rate in comparison to non-modified enzyme that results in a lesser extent of crosslinking of said substrate polymer in comparison to a matrix formed with non-modified enzyme.
A cross-linked matrix, comprising a substrate polymer and a modified enzyme molecule, wherein said substrate polymer has been crosslinked by said modified enzyme molecule, wherein both said modified enzyme and said substrate polymer are present in the matrix, said modified enzyme molecule having a modification that modifies an electrostatic charge of said modified enzyme molecule to be of opposite sign to a net charge of said substrate polymer by changing the isoelectric point (pI) of said modified enzyme in comparison to unmodified enzyme, wherein said modification reduces the crosslinking rate in comparison to non-modified enzyme resulting in a lesser extent of crosslinking of said substrate polymer in comparison to a matrix formed with non-modified enzyme.
A cross-linked matrix, comprising a substrate polymer and a modified enzyme molecule, wherein said substrate polymer has been crosslinked by said modified enzyme molecule, wherein both said modified enzyme and said substrate polymer are present in the matrix, said modified enzyme molecule having a modification such that said modified enzyme molecule has a reduced cross-linking rate in comparison to non-modified enzyme that results in a lesser extent of crosslinking of said substrate polymer in comparison to a matrix formed with non-modified enzyme, wherein said reduced cross-linking rate is at least 10% of the non-modified enzyme cross-linking rate.
A cross-linked matrix, comprising a substrate polymer and a modified enzyme molecule, wherein said substrate polymer has been crosslinked by said modified enzyme molecule, wherein both said modified enzyme and said substrate polymer are present in the matrix, said modified enzyme molecule being modified by applying an activated PEG to an enzyme molecule and thereby producing a PEGylated enzyme molecule as said modified enzyme molecule, wherein said activated PEG comprises methoxy PEG (mPEG), its derivatives, mPEG-NHS, succinimidyl (NHS) esters of mPEG (mPEG-succinate-NHS), mPEG-glutarate-NHS, mPEG-valerate-NHS, mPEG-carbonate-NHS, mPEG-carboxymethyl-NHS, mPEG-propionate-NHS, mPEG-carboxypentyl-NHS), mPEG-nitrophenylcarbonate, mPEG-propylaldehyde, mPEG-Tosylate, mP EG-carbonylimidazole, mPEG-isocyanate or mPEG-epoxide, wherein said modification reduces the crosslinking rate in comparison to non-modified enzyme resulting in a lesser extent of crosslinking of said substrate polymer in comparison to a matrix formed with non-modified enzyme .
A method for controlling formation of a matrix, comprising
- modifying an enzyme molecule with a modification selected from the group consisting of:
- a modification that increases the actual size of said enzyme molecule;
- or
- a modification that increases the hydrodynamic volume of said modified enzyme molecule;
- mixing said modified enzyme molecule with at least one substrate polymer that is a substrate of said modified enzyme molecule;
- and
- forming the matrix through crosslinking of said at least one substrate polymer by the action of said modified enzyme molecule,
- wherein said modification reduces the crosslinking rate in comparision to non-modified enzyme resulting in a lesser extent of crosslinking of said substrate polymer in comparison to a matrix formed with non-modified enzyme, and
- wherein said forming the matrix is at least partially controlled by said modification of said enzyme molecule and wherein
- said matrix comprises said modified enzyme molecule and cross-linked substrate polymer.
Dependencies are inferred from the claim text.