Methods for extracting keratin proteins
Described herein are methods to produce keratin protein-based biomaterials, the parameters required to achieve improved extraction, the parameters required to improve isolation, the parameters of lyophilization and the grinding process to achieve consistent particulate sizes of protein materials.
1. A method for extracting keratin proteins comprising:
a. Treating a keratin protein source with an oxidizing or reducing agent to solubilize keratin proteins;
b. Separating the soluble proteins from the keratin protein source by high speed centrifugation to produce a clarified soluble keratin protein solution; and
c. Dialyzing the clarified soluble keratin protein solution to obtain a soluble keratin protein solution which exhibits a conductivity of 0.6 mS or less,
wherein said keratin protein is keratose.
2. The method of claim 1 , wherein said high speed centrifugation is performed at 5,000 rpm or higher.
3. The method of claim 1 , wherein said clarified soluble keratin protein solution comprises less than 5% beta-keratin.
4. The method of claim 1 , wherein said method comprises dialyzing the clarified soluble keratin protein solution against a 5 kDa molecular weight membrane.
5. The method of claim 1 , wherein said dialysis is performed at a pressure from about 10 psi to about 70 psi.
6. The method of claim 5 , wherein the dialysis permeate comprises gamma-keratose.
7. The method of claim 1 , wherein the soluble keratin protein solution is lyophilized into a keratin protein cake.
8. The method of claim 1 , wherein said soluble keratin protein solution comprising at least 90% or more keratose.
9. The method of claim 8 , wherein said soluble keratin protein solution comprises at least 90% or more alpha-keratose.
10. A method for extracting keratin proteins comprising:
a. Treating a keratin protein source with an oxidizing or reducing agent to solubilize keratin proteins;
b. Separating the soluble proteins from the keratin protein source by high speed centrifugation to produce a clarified soluble keratin protein solution; and
c. Dialyzing the clarified soluble keratin protein solution to obtain a soluble keratin protein solution which exhibits a conductivity of 0.6 mS or less,
wherein said soluble keratin protein solution comprising at least 90% or more keratose.
11. A method for extracting keratin proteins comprising:
a. Treating a keratin protein source with an oxidizing or reducing agent to solubilize keratin proteins;
b. Separating the soluble proteins from the keratin protein source by high speed centrifugation to produce a clarified soluble keratin protein solution; and
c. Dialyzing the clarified soluble keratin protein solution to obtain a soluble keratin protein solution which exhibits a conductivity of 0.6 mS or less,
wherein said dialysis is performed at a pressure from about 10 psi to about 70 psi.
12. The method of claim 11 , wherein said keratin protein is kerateine.