Methods for producing, films comprising, and methods for using heterogenous crosslinked protein networks

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising:

• treating soluble α-keratin proteins comprising disulfide bonds under first conditions effective to break said disulfide bonds and to convert cystine residues to thiolate anions;
• exposing said thiolate anions to a crosslinking agent comprising at least a first functional group and a second functional group reactive with thiolate anions under conditions effective to induce a first reaction between said first functional group on a plurality of molecules of said crosslinking agent and first thiolate anions on a plurality of first α-keratin protein molecules, and to induce a second reaction between said second functional group on a plurality of molecules of said crosslinking agent and second thiolate anions on a plurality of second α-keratin proteins, thereby producing a heterogeneous cross-linked keratin network.

A method for functionalizing α-keratin proteins comprising exposing said α-keratin proteins comprising reactive pendant groups to a nucleophilic substitution agent comprising at least one terminal epoxide under conditions effective to induce a plurality of said reactive pendant groups to react with said nucleophilic substitution agent, thereby producing a plurality of epoxidized α-keratin molecules.

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising exposing α-keratins comprising reactive pendant groups to a crosslinking agent comprising at least a first terminal epoxide and a second terminal epoxide under conditions effective to induce a first reaction between said first terminal epoxide on a plurality of molecules of said crosslinking agent and first reactive pendant groups on a plurality of first α-keratin proteins molecules, and to induce a second reaction between said second terminal epoxides on a plurality of molecules of said crosslinking agent and second reactive pendant groups on a plurality of second α-keratin proteins molecules, thereby producing a heterogeneous cross-linked keratin network.

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising exposing soluble α-keratin proteins comprising reactive pendant groups to a crosslinking agent comprising a plurality of carboxylic acid groups under conditions effective to induce first reactions between first carboxylic acid groups on a plurality of molecules of said crosslinking agent and first reactive pendant groups on a plurality of first α-keratin protein molecules, and to induce second reactions between second carboxylic acid groups on a plurality of molecules of said crosslinking agent and second reactive pendant groups on a plurality of second α-keratin protein molecules, thereby producing a heterogeneous cross-linked keratin network.

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising exposing soluble α-keratin proteins comprising reactive pendant groups to phthallic anhydride under conditions effective to induce a first reaction between first reactive pendant groups on a plurality of first α-keratin protein molecules and first carbox-oyl groups of a plurality of molecules of said phthallic anhydride, and to induce a second reaction between second reactive pendant groups on a plurality of second α-keratin protein molecules and second carbox-oyl groups of a plurality of molecules of said phthallic anhydride, thereby producing a heterogeneous cross-linked keratin network.

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising exposing soluble α-keratin proteins comprising reactive pendant groups to a crosslinking agent comprising a plurality of cyanate groups under conditions effective to induce a first reaction between first cyanate groups on a plurality molecules of said crosslinking agent and first reactive pendant groups on a plurality of first α-keratin proteins molecules, and to induce a second reaction between second cyanate groups on a plurality of molecules of said crosslinking agent and second reactive pendant groups on a plurality of second α-keratin protein molecules, thereby producing a heterogeneous cross-linked keratin network.

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising exposing soluble α-keratin proteins comprising pendant thiol groups to phthallic anhydride under conditions effective to induce a first reaction between a first thiol group on a plurality of first α-keratin proteins molecules and first carbox-oyl groups of a plurality of said phthallic anhydride molecules, and to induce a second reaction between second thiol groups on a plurality of second α-keratin protein molecules and second carbox-oyl groups of a plurality of said phthallic anhydride molecules, thereby producing a heterogeneous cross-linked keratin network.

A method for making a keratin network comprising a heterogeneous crosslinking agent, said method comprising exposing α-keratin proteins comprising carboxylic acid pendant groups to a crosslinking agent comprising at least a first nucleophilic group and a second nucleophilic group under conditions effective to induce a first reaction between said first nucleophilic group on a plurality of molecules of said crosslinking agent and first carboxyl group on a plurality of first α-keratin protein molecules, and to induce a second reaction between said second nucleophilic group on a plurality of molecules of said crosslinking agent and second carboxylic acids on a plurality of second α-keratin protein molecules, thereby producing a heterogeneous cross-linked keratin network.

A method for making a protein network comprising a heterogeneous crosslinking agent, said method comprising exposing soluble proteins comprising reactive pendant groups to a heterogeneous crosslinking agent comprising a first functional group and a second functional group adapted to react with said reactive pendant groups under conditions effective to induce a first reaction between said first functional groups on a plurality of molecules of said crosslinking agent and first reactive pendant groups on a plurality of first soluble protein molecules, and to induce a second reaction between said second functional groups on a plurality of molecules of said crosslinking agent and second reactive pendant groups on a plurality of second soluble protein molecules, thereby producing a heterogeneous cross-linked protein network.

A heterogeneous crosslinked protein network comprising a plurality of protein molecules interlinked by a crosslinking agent, said network comprising first bonds between first functional groups on a plurality of molecules of said crosslinking agent and first pendant groups on a plurality of first protein molecules and second bonds between second functional groups on a plurality of molecules of said crosslinking agent and second reactive pendant groups on a plurality of second protein molecules.

A heterogeneous crosslinked keratin network comprising a plurality of α-keratin molecules interlinked by a crosslinking agent, said network comprising first bonds between first functional groups on a plurality of molecules of said crosslinking agent and first pendant groups on a plurality of first protein molecules and second bonds between second functional groups on a plurality of molecules of said crosslinking agent and second reactive pendant groups on a plurality of second protein molecules.

A heterogeneous crosslinked proteinaceous network comprising the following crosslinks:

• 38
• wherein R1 and R2 independently are amino acid residues from separate protein molecules, the residues being selected from the group consisting of cysteine, arginine, serine, lysine, asparagine, glutamine, tyrosine, tryptophan, and histidine.

A heterogeneous cross-linked proteinaceous network comprising the following crosslinks:

• 39
• wherein R1 and R2 independently are amino acid residues from separate protein molecules, the residues being selected from the group consisting of cysteine, arginine, serine, lysine, asparagine, glutamine, tyrosine, tryptophan, and histidine.

A method for increasing or decreasing the dissolution rate of a heterogeneous crosslinked proteinaceous network comprising controlling a quantity of crosslink bonds having a characteristic selected from the group consisting hydrophobicity, hydrolytic stability, and steric hindrance.