Biomaterials and methods related thereto

A method for producing a crosslinked biopolymer composition, the method comprising:

• providing a composition comprising:
• a. an α-crystallin;
• b. a β-crystallin;
• c. a γ-crystallin;
• d. a protein from any one of a) to c) above from Hoki (Macruronus novaezelandiae);
• e. a protein from any one of a) to c) above from Homo sapiens;
• f. a protein comprising the amino acid sequence identified in Table 1 herein;
• g. a polypeptide comprising or consisting of at least about 10 contiguous amino acids from any one of a) to f) above;
• h. a protein having at least about 90% amino acid identity to any one of a) to g) above;
• i. a protein according to any one of a) to h) above having the native structure of a crystallin protein in vivo;
• j. any combination of two or more of a) to i) above;
• optionally one or more plasticizers;
• optionally one or more co-initiators;
• and
• contacting said composition with one or more crosslinking molecules;
• initiating crosslinking, thereby forming a crosslinked biopolymer composition.

A method for producing a composition comprising one or more purified crystallin proteins, the method comprising:

• providing vertebrate eye tissue;
• homogenising the tissue in the presence of an extraction buffer under conditions suitable for the maintenance of native crystallin protein structure;
• separating the liquid homogenate from any residual solids, for example by centrifugation or filtration, to provide a crystallin protein-containing solution;
• optionally at least partially further purifying the crystallin protein;
• optionally dialysing the crystallin protein-containing solution to remove the extraction buffer;
• optionally lyophilising the crystallin protein-containing solution to provide a lyophilised crystallin protein composition;
• optionally storing the crystallin protein-containing solution or the lyophilised crystallin protein composition, for example at or below 0° C.;
• wherein a substantial proportion of the purified crystallin proteins retain their native structure.

A method of tissue closure in a subject in need thereof, the method comprising

• optionally applying force to close the laceration, lesion, incision or wound;
• contacting a laceration, lesion, incision, or wound or the site of said laceration, lesion, incision, or wound with a crystallin protein containing composition as defined in any one of the preceding claims, optionally wherein the crystallin protein containing composition is at least partially crosslinked,
• optionally applying force to close the laceration, lesion, incision or wound,
• initiating and/or maintaining crosslinking;
• maintaining the closure of the laceration, lesion, incision or wound for a time sufficient for crosslinking to occur;
• wherein crosslinking of the crystallin proteins forms an adhesive composition.

A method of tissue closure in a subject in need thereof, wherein the subject is undergoing or who has undergone ophthalmic surgery, the method comprising

• contacting a surgical incision or the site of said surgical incision with a crystallin protein containing composition as defined in any one of the preceding claims, optionally wherein the crystallin protein containing composition is at least partially crosslinked;
• optionally applying force to close the incision;
• initiating and/or maintaining crosslinking;
• maintaining the closure of the surgical incision for a time sufficient for crosslinking to occur;
• wherein crosslinking of the crystallin proteins forms an adhesive composition capable of maintaining closure of the surgical incision.

A method of treating an ocular injury or ocular incision in a subject in need thereof, the method comprising the steps:

• contacting the ocular injury or incision with a composition as described herein, optionally wherein the crystallin protein containing composition is at least partially crosslinked;
• and
• initiating and/or maintaining crosslinking;
• wherein the crosslinking forms a bioadhesive polymer composition.

A method of delivering one or more active agents to a subject in need thereof, the method comprising

• providing a crystallin protein comprising composition as defined in any one of the preceding claims, optionally wherein the crystallin protein containing composition is at least partially crosslinked, wherein the composition additionally comprises one or more active agents,
• contacting the subject with the composition,
• optionally initiating and/or maintaining crosslinking of the composition,
• thereby delivering the active agent to the subject in need thereof.

The composition, method or use of any one of the preceeding claims, wherein

• a. the native secondary structure of the one or more crystallin proteins is maintained;
• or
• b. the native tertiary structure of the one or more crystallin proteins is maintained;
• or
• c. the native quaternary structure of the one or more crystallin proteins is maintained;
• or
• d. the one or more crystallin proteins are substantially free of nanofibrils or other disrupted structural forms;
• or
• e. at least some of the crystallin protein present in the composition is natively glycosylated;
• or
• f. any combination of two or more of a) to e) above.

The composition, method or use of any one of the preceeding claims, wherein the composition comprises

• a. from about 0.1% w/w to about 1.5% w/w crosslinker;
• or
• b. from about 0.5% w/w to about 3% w/w crosslinker;
• or
• c. from about 3% w/w to about 30% w/w crosslinker;
• or
• d. from about 10 mg/mL to about 200 mg/mL crystallin protein;
• or
• e. from about 10 mg/mL to about 120 mg/mL crystallin protein;
• or
• f. from about 0.5% w/w to about 3% w/w plasticizer;
• or
• g. from about 0.5% w/w to about 5% w/w co-initiator;
• or
• h. any combination of two or more of a) to g) above.

The composition, method or use of any one of the preceeding claims, wherein the composition comprises

• a. from about 100 mg/mL to about 120 mg/mL crystallin protein, from about 5 mM to about 10 mM glutaraldehyde, and from about 1.5% w/w to about 2.5% w/w glycerol;
• or
• b. from about 100 mg/mL to about 120 mg/mL crystallin protein, from about 10% w/w to about 20% w/w PEGDA, and from about 0.2% w/w to about 1.0% w/w photoinitiator;
• or
• c. from about 50 mg/mL to about 80 mg/mL crystallin protein, from about 10% w/w to about 20% w/w PEGDA, and from about 0.2% w/w to about 1.0% w/w photoinitiator;
• or
• d. from about 50 mg/mL to about 80 mg/mL crystallin protein, from about 25% w/w to about 50% w/w PEGDA, from about 0.2% to about 1.0% w/w photoinitiator, and from 10% w/w to 20% w/w of co-initiator.

The composition, method or use of any one of the preceeding claims, wherein the composition when crosslinked

• a. is optically transparent over the visible spectrum;
• or
• b. has a refractive index equivalent to that of the eye of a subject to whom it is or has been administered;
• or
• c. has a transmittance of light across the visible spectrum (400 nm to 700 nm) of greater than about 75%;
• or
• d. any combination of two or more of a) to c) above.