Methods for separating and purifying endogenous, exogenous and recombinant proteins/peptides from plants and animals using aqueous-free, anhydrous strategies

Abstract

The present invention relates to recombinant proteins/peptides from plant and animal materials, compositions comprising the proteins/peptides and methods for making them.

What is claimed is: 
     
         1 . A method for obtaining a recombinant protein or peptide from plant or animal material after its expression inside the host as a fusion protein, wherein the fusion protein comprises a protein or peptide of interest, a carrier protein and a gas/vapor phase cleavage site situated between the protein or peptide of interest and the carrier protein comprising:
 (a) incubating the plant or animal material with gas or vapors to cleave and release the protein or peptide of interest from the fusion carrier, wherein the plant or animal material has been naturally dried or desiccated, or dried to 25 percent moisture content or less; and   (b) separating the released protein or peptide from its carrier and the remaining plant or animal material.   
     
     
         2 . The method of  claim 1 , wherein the fusion carrier causes the localization and deposition of the fusion protein on to the surface of a plant or animal organelle or structural component. 
     
     
         3 . The method of  claim 2 , wherein the plant or animal organelle or structural component is including but not limited to a starch granule, protein body, cell wall, chloroplast membrane, flagella or eye spot. 
     
     
         4 . The method of  claim 3 , wherein the plant or animal organelle or structural component is a starch granule, cell wall or protein body. 
     
     
         5 . The method of  claim 4 , wherein the plant or animal organelle or structural component is a starch granule. 
     
     
         6 . The method of  claim 1 , wherein the biological organelle or structural component is separated from other cellular components using milling and air-classification technologies prior to incubation with a cleavage inducing gas or vapor. 
     
     
         7 . The method of  claim 1 , wherein the gas or vapor cleavable linker sequence is Asp-Pro, Met, Ser, Trp, Asn-Gly, Gly-Gly or Gly-Thr. 
     
     
         8 . The method of  claim 7 , wherein the gas or vapor cleavable linker sequence is Asp-Pro, Gly-Gly or Gly-Thr. 
     
     
         9 . The method of  claim 8 , wherein the gas or vapor cleavable linker sequence is Asp-Pro. 
     
     
         10 . The method of  claim 9 , wherein the Asp-Pro peptide linker sequence is cleaved using a gas or vapor composed from but not limited to: heptafluorobutyric acid, acetic acid, perfluorobutyric acid, hydrochloric acid, formic acid, trifluoroacetic acid or perfluoric acid. 
     
     
         11 . The method of  claim 10 , wherein the Asp-Pro peptide linker sequence is cleaved using heptafluorobutyric acid gas or vapor. 
     
     
         12 . The method of  claim 11 , wherein the concentration of heptafluorobutyric acid is from 0.05 percent to 20 percent. 
     
     
         13 . The method of  claim 12 , wherein the concentration of heptafluorobutyric acid is 0.1 percent to 5 percent. 
     
     
         14 . The method of  claim 11 , wherein the reaction temperature is from 25 degrees Celsius to 100 degrees Celsius. 
     
     
         15 . The method of  claim 14 , wherein the reaction temperature is from 40 degrees Celsius to 80 degrees Celsius. 
     
     
         16 . The method of  claim 15 , wherein the reaction temperature is from 55 degrees Celsius to 65 degrees Celsius. 
     
     
         17 . The method of  claim 11 , wherein the incubation time of the reaction is from 1 hour to 60 hours. 
     
     
         18 . The method of  claim 17 , wherein the incubation time of the reaction is from 10 hours to 40 hours. 
     
     
         19 . The method of  claim 18 , wherein the incubation time of the reaction is from 14 hours to 18 hours. 
     
     
         20 . The method of  claim 1 , wherein the plant or algal organelle or structural component is isolated using milling and/or air cyclone and/or air-classification technologies prior to gas/vapor exposure. 
     
     
         21 . The method of  claim 1 , wherein the protein or peptide of interest is trapped/captured from an air flow using a filter device following gas/vapor phase cleavage. 
     
     
         22 . The method of  claim 21 , wherein the filter device is composed of but not limited to: glass fibers, quartz microfibers, polybrene or cellulose. 
     
     
         23 . The method of  claim 1 , wherein the protein or peptide of interest is trapped/captured from an air flow using a dust collection device following gas/vapor phase cleavage. 
     
     
         24 . The method of  claim 1 , wherein the protein or peptide of interest is trapped/captured from an air flow following gas/vapor phase cleavage using an electrostatic surface-trapping device of known art in the powder control technologies field.