Cell-free protein synthesis platform

Abstract

Provided herein in various embodiments is a cell-free protein synthesis platform, comprising a ribosome and a polymer membrane comprising a polymer membrane pore and an affinity binding site capable of binding the ribosome, wherein the polymer membrane separates a reaction compartment from a product compartment. Different implementations and modified ribosomes are used in various embodiments.

Description

CELL-FREE PROTEIN SYNTHESIS PLATFORMCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is an international patent application filed in accordance with the Patent Cooperation Treaty. This application claims the priority benefit of U.S. Provisional Patent Application Nos. 63 / 570,427, filed 27 March 2024, and entitled “CELL-FREE PROTEIN SYNTHESIS PLATFORM”. The disclosure of U.S. Provisional Patent Application No. 63 / 570,427 is hereby incorporated herein by reference in its entirety.REFERENCE TO SEQUENCE LISTING

[0002] This application contains a Sequence Listing that has been submitted electronically herewith as an XML file named VST-0001-PV_SequenceListing_26March2024.xml. The XML file, created on March 26, 2024, has a size of 486,225 bytes. The contents of the Sequence Listing XML file are incorporated herein by reference in their entirety.BACKGROUND

[0003] Cell-free protein synthesis is a method by which proteins are expressed by utilizing an extract of molecular machinery. This extract contains ribosomes, tRNA molecules, energy molecules and other associated proteins in addition to the genetic material for the protein of interest. Cell-free protein synthesis is a very broad field of study within biochemistry and was used to deduce the genetic code. Cell-free protein synthesis is in essence a system that can produce proteins using cellular machinery outside of a biological cell.SUMMARY

[0004] In some embodiments, provided herein is a cell-free protein synthesis platform, comprising a ribosome and a polymer membrane comprising a polymer membrane pore and an affinity binding site capable of binding the ribosome, wherein the polymer membrane separates a reaction compartment from a product compartment.

[0005] In some embodiments, the ribosome is bound to the polymer membrane. In some embodiments, the ribosome is bound to the polymer membrane by a membrane bound protein embedded within the polymer membrane. In some embodiments, the membrane bound protein isa translocon. In some embodiments, the membrane bound protein is a hinge protein. In some embodiments, the membrane bound protein comprises a translocon embedded in the polymer membrane, bound to a hinge protein.

[0006] In various embodiments provided herein, the cell-free protein synthesis platform comprises a reaction solution. In some embodiments, the reaction solution comprises a cell-free protein extract. In some embodiments, the cell-free protein extract comprises an excess of small ribosomal subunits. In various embodiments, the reaction solution or cell-free protein extract comprises a molar excess of small ribosomal subunits. In various embodiments, there are significantly more small ribosomal subunits as compared with bound large ribosomal subunits. This can, in various embodiments, provide for the small ribosomal subunits being present in the bulk reaction solution as compared with large ribosomal subunits bound to the polymer membrane.

[0007] In some embodiments, the cell-free protein synthesis platform further comprises a housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compailment. In some embodiments, the cell-free protein synthesis platform further comprises a polymer membrane bag having an inside membrane surface and an outside membrane surface. In some embodiments the affinity binding site is on the inside membrane surface of the polymer membrane bag. In some embodiments the affinity binding site is on the outside surface of the polymer membrane bag. In some embodiments, the polymer membrane bag is a dialysis bag. In some embodiments the housing comprises a polymer membrane tube, cap and base attached to the bottom of the polymer membrane tube. In some embodiments, the tube cap further comprises a hole for reagent addition. In some embodiments, polymer tube cap and base are further housed in a container. In some embodiments, the protein is translated through the polymer membrane tube into the container housing. In some embodiments, the protein is translated from the reaction solution in the housing through the polymer membrane into the polymer membrane tube.

[0008] In some embodiments of the cell-free protein synthesis platform provided herein, the ribosome is a modified ribosome. In some embodiments, the modified ribosome comprises one or more modified ribosome sites. In some embodiments, the modified ribosome comprises 1 to 10 ribosome ligand binding sites. In some embodiments the ribosome comprises 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 ligand binding sites. In some embodiments, the modifiedribosome comprises a ribosome ligand binding site. In some embodiments, the modified ribosome comprises two or more ribosome ligand binding sites. In some embodiments, the ribosome ligand binding sites are different ligand binding sites. In some embodiments, the ribosome ligand binding site is a molecular' binding site. In some embodiments, the ribosome ligand binding site is an affinity molecule binding site. In some embodiments, the ribosome ligand binding site is an antibody epitope. In some embodiments, the ribosome ligand binding site is selected from His-tag, His-Glu-tag, Asp-Lys-tag, Glu-tag, Arg-tag, Cys-tag, FLAG-tag, Capture Select C-tag, TC-tag, Xpress-tag, Softag-3-tag, Strep-tag, Aul, AU5, T7-tag, V5-tag, B- tag, E2 epitope, HSV epitope, KT3 epitope, Myc epitope, PDZ ligands, Asp-tag, Phe-tag, Protein C, Sl-tag, Universal-tag, VSV-G, a HTTPHH, functional equivalents, or combinations thereof. In some embodiments, the ribosome ligand binding site is on a ribosomal protein. In some embodiments, the ribosomal protein comprising the ribosome ligand binding site is from a different organism than the ribosome. In some embodiments, the ribosomal protein ligand binding site further comprises a recombinant ribosome. In some embodiments of the cell-free protein synthesis platform provided herein, the ribosomal protein comprising the ribosome ligand binding site is modified from its native form.

[0009] In some embodiments, the ribosome ligand binding site is on a ribosomal RNA. In some embodiments, the ribosomal RNA comprising the ribosome ligand binding site is from a different organism than the ribosome. In some embodiments, the ribosomal RNA comprising the ribosome ligand binding site is ribosomal RNA modified from its native form. In some embodiments, the ribosome ligand binding site is a naturally occurring ribosomal RNA ligand binding site.

[0010] In some embodiments of the cell-free protein synthesis platform provided herein, the affinity binding site of the polymer membrane comprising an affinity binding site further comprises an affinity molecule. In some embodiments, the affinity molecule is capable of binding a ribosome ligand binding site. In some embodiments, the affinity binding site comprises an affinity molecule that binds a ribosome ligand binding site.

[0011] In some embodiments of the cell-free protein synthesis platform provided herein, the ribosome is attached to the polymer membrane. In some embodiments, the ribosome is bound to one or more affinity molecules on the polymer membrane. In some embodiments, the ribosome is bound to 1 to 10 affinity molecules on the polymer membrane. In some embodiments, theribosome is bound to 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 affinity molecules on the polymer membrane. In some embodiments, the ribosome contains a naturally occurring ribosome ligand binding site. In some embodiments, the ribosome ligand binding site is capable of binding an affinity molecule at the affinity binding site. In various embodiments, the affinity molecule is bound near the polymer membrane pore. In some embodiments, the affinity molecule is on the polymer membrane pore edge. In some embodiments, the affinity molecule is 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5-50 nm, 0.5-25 nm, 0.5 -10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-50 nm, 5-25 nm, or 5-10 nm from the polymer membrane pore edge. In various embodiments the affinity molecule is selected from a metal chelating tag, an antibody epitope, a covalent binding molecule, and functional equivalents and combinations thereof. In some embodiments, the affinity molecule is selected from HAT-tag, His-tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1 , AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl-Sepharose (Phe-tag), ProC, SplAsH (Cys- tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA-aptamer, RNA- Sephadex, RNA-Streptavadin), tags made by the SELEX process, functional equivalents, and combinations thereof. In some embodiments, each polymer membrane pore has 1 to 10 affinity binding molecules within 100 nm of the edge of the polymer membrane pore. In some embodiments each polymer membrane pore has 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 affinity binding molecules within 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5-50 nm, 0.5-25 nm, 0.5-10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-50 nm, 5-25 nm, or 5-10 nm from the polymer membrane pore edge. In some embodiments, the cell-free protein synthesis platform uses two or more different affinity molecules. In some embodiments, the cell-free protein synthesis platform uses 1-3, 1-5, 1-10, 2- 5, 2-10, 3-5, 3-10, 4-6, or 4-10 different affinity molecules.

[0012] In various embodiments of the cell-free protein synthesis platform provided herein, the modified ribosome is attached to the polymer membrane. In some embodiments, a naturally occurring ribosome with naturally occurring ligand binding sites is attached to the polymer membrane.

[0013] In various embodiments of the cell-free protein synthesis platform provided herein theribosome is a reconstituted ribosome, a recombinant ribosome, or a hybrid ribosome.

[0014] In various embodiments of the cell-free protein synthesis platform provided herein the ribosome ligand binding site is attached via one or more intermediate molecules to the affinity molecule attached to the polymer membrane. In some embodiments, the intermediate molecule is a signal recognition protein. In some embodiments, the signal recognition peptide is a modified signal recognition peptide. In some embodiments, the modified signal recognition protein is selected from SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259 SEQ ID NO: 260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO: 269, SEQ ID NO: 270, SEQ ID NO: 271, functional equivalents, or combinations thereof. In some embodiments, the intermediate molecule is a polymer, antibody, antibody complex, small molecule chemical linker, hinge protein, dendrimer, functional equivalents, or combinations thereof. In some embodiments, ribosome ligand binding site is attached via 2-10 intermediate molecules to the affinity molecule on the polymer membrane. In some embodiments, the ribosome ligand binding site is attached via 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 intermediate molecules to the affinity molecule on the polymer membrane.

[0015] In some embodiments of the cell-free protein synthesis platform provided the hybrid ribosome comprises ribosomal proteins and RNA selected from two or more organisms. In some embodiments, the hybrid ribosome comprises ribosomal proteins and RNA selected from archaea, eukaryotes, prokaryotes or combinations thereof.

[0016] In some embodiments of the cell-free protein synthesis platform provided herein the ribosome comprises a modified ribosomal protein. In some embodiments, the modified ribosomal protein comprises modifications selected from amino acid substitution, protein tags, non-canonical amino acid substitution, functional equivalents, and combinations thereof. In some embodiments, the modified ribosomal protein is selected from SEQ ID NO:1 to 256, SEQ ID NO: 272 to SEQ ID NO:490, functional equivalents, or combinations thereof. In various embodiments, combinations of modified ribosomal proteins can be used. In some embodiments, the modified ribosomal proteins are prepared using cell-free protein synthesis. In some embodiments, the modified ribosomal proteins are prepared using various embodiments of the platform described herein.

[0017] In some embodiments of the cell-free protein synthesis platform provided herein theribosome ligand binding site is a naturally occurring ribosomal RNA sequence. In some embodiments, the ribosome ligand binding site is a modified ribosomal RNA sequence.

[0018] In some embodiments of the cell-free protein synthesis platform provided herein the affinity molecule is an RNA tag. In some embodiments, the RNA tag is selected from RNA- biotin, RNA-aptamer, RNA-Sephadex, RNA-Streptavadin„ functional equivalents, and combinations thereof. In some embodiments, a ribosomal RNA ligand binding site binds the affinity molecule on a polymer membrane. In some embodiments, one or more ribosomal RNA ligand binding sites can bind one or more affinity molecules near a membrane pore. In some embodiments, 1 to 10 ribosomal RNA ligand binding sites can bind 1 to 10 affinity molecules near a membrane pore. In some embodiments, 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 ribosomal RNA ligand binding sites can bind 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 affinity molecules on the polymer membrane.

[0019] In some embodiments of the cell-free protein synthesis platform provided herein, the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules and / or energy systems, buffers, salts. In various embodiments, the reaction solution further comprises an excess of small ribosomal subunits. In some embodiments, the reaction solution further comprises an mRNA for a protein of interest. In some embodiments, the reaction solutions comprise a cell free protein extract. In some embodiments, the cofactors in the cell free protein synthesis extract comprise proteins, ions, and RNA. In some embodiments, the tRNA’s comprise naturally occurring, modified and synthesized tRNA molecules. In some embodiments, amino acids are selected from naturally occurring amino acids, noncanonical amino acids, functional equivalents, and combinations thereof. In some embodiments, amino acids are selected from alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine, pryrrolysine (stop codon amino acids), homoallylglycine, homopropargylglycine, azidohomoalanine, azidonorleucine, azido phenylalanine, acetyl phenylalanine, acetyl phenylalanine, propargyloxyphenylalanine, functional equivalents, and combinations thereof. In some embodiments, the energy molecules and / or energy systems comprise creatine phosphate / creatine kinase, glucose, glutamate decarboxylase, maltodextrin, nucleoside triphosphates (NTPs), phosphoenolpyruvate, 3-phosphoglycerate, functional equivalents and combinations thereof. In some embodiments, the reaction solution componentsare prepared using cell-free protein synthesis. In some embodiments, the reaction solution components are prepared using various embodiments of the platform described herein.

[0020] In some embodiments of the cell-free protein synthesis platform provided herein the polymer membrane is comprised of monomers or polymers selected from acrylamide, 3- acrylamidopropyl trimethylammonium chloride, N-hydroxymethyl acrylamide, N,N- methylenebisacrylamide, cellulose acetate, N,N-dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n-butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, polysacrylamido-2- methyl-1 -propane-sulfonic acid, poly aery lie acid, poly methacrylic acid, polyisopropylacrylamide, poly-3-acrilamidopropyl trimethylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polydiallyldimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide, isopropylacrylamide, or vinylpyrrolidone, copolymers of acrylamide-2-methyl-lpropanesulfonic acid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido-propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl-pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidone, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfide, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl-ammonium, polyvinyl chloride, poly vinylidene fluoride, poly-4-vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinylsulfonic acid, functional equivalents and combinations thereof. In some embodiments, the polymer membrane material is selected from the more commonly used industrial polymers, polyamide, polyacrylonitrile, polyether sulfone, polyvinylidene fluoride, functional equivalents, and combinations thereof. In some embodiments, the polymer membrane is a hydrogel membrane.

[0021] In some embodiments, the polymer membrane has a thickness of at least Inm, 5 nm, 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 50 nm, 75 nm, or 100 nm. In some embodiments the polymermembrane has a thickness selected from of 1-100 nm, 1-75 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-75 nm, 5-50 nm, 5-25 nm, 5-10 nm, 10-100 nm, 10-75 nm, 10-50 nm, 10-25 nm, 25-100 nm, 25-75 nm, or 25-50 nm. In some embodiments, the polymer membrane has a thickness of 5 nm to 10 nm. In some embodiments, the polymer membrane is comprised of one or more layers. In some embodiments, the polymer membrane is comprised of 1-2, 1-3, 1-5, 1- 10, 2-5, 2-10, or 5-10 layers. In some embodiments, the polymer membrane comprises one or more layers of a polymer support matrix. In some embodiments, the polymer membrane comprises 1-2, 1-3, 1-5, 1-10, 2-5, 2-10, or 5-10 layers of a polymer support matrix.

[0022] In some embodiments the layers of polymer support matrix increase in dimensions and / or decrease in density to provide increasing permeability of the outer layers. In some embodiments, polymer support matrix layers increase structural integrity of the polymer membrane. In some embodiments the polymer membrane support matrix is comprised of monomers or polymers selected from the monomers and polymer that can comprise the polymer membrane described herein.

[0023] In some embodiments, the polymer membrane is a semi permeable membrane. In some embodiments, the polymer membrane is selectively permeable to water and ions of interest. In some embodiments, the polymer membrane pores have a diameter of about 3 nm to about 10 nm. In some embodiments, the polymer membrane pores have a diameter selected from 5-100 nm, 5- 75 nm, 5-50 nm, 5-25 nm, 5-10 nm, 10-100 nm, 10-75 nm, 10-50 nm, 10-25 nm, 20-100 nm, 20- 75 nm, 20-50 nm, 20-25 nm, 50-100 nm, or 50-75 nm. In some embodiments, the polymer membrane pores are spaced at least 10 nm, 20 nm, 25 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, or 500 nm edge to edge apart from other membrane pores. In some embodiments, the polymer membrane comprises 1-10 affinity molecules within 10 nm of the edge of each membrane pores. In some embodiments the polymer membrane comprises 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 affinity molecules within 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5-50 nm, 0.5- 25 nm, 0.5-10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-50 nm, 5-25 nm, or 5-10 nm from the polymer membrane pore edge. In some embodiments, the polymer membrane comprises 1-6 affinity molecules within 10 nm of the edge each membrane pores.

[0024] In some embodiments, the polymer membrane comprises one or more coatings. In someembodiments, the polymer membrane comprises 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 layers of coatings. In some embodiments, the coating material is selected form the list of monomers and polymers that comprise the polymer membrane listed herein. In some embodiments, the coating comprises cellulose or other functionally equivalent membrane coating. In some embodiments, the polymer membrane comprises one or more embedded membrane proteins. In some embodiments the embedded membrane protein is a translocon. In some embodiments, the translocon is a complex of proteins. In some embodiments, the translocon is bound to a modified ribosome. In some embodiments, the translocon is bound to a hinge protein, intermediate molecule, antibody, antibody complex, dendrimer, functional equivalents, and combinations thereof. In some embodiments pore dimension and dispersion during manufacturing are controlled by having dendrimers embedded within the polymer membrane. In various embodiments, dendrimers are included in the membrane manufacturing process along with other components. In some embodiments, the dendrimers are removed from the polymer membrane during manufacturing. In some embodiments, dendrimers are dissolved from the membrane.

[0025] In some embodiments of the cell-free protein synthesis platform provided herein, a hinge protein, intermediate molecule, antibody, antibody complex, dendrimer, functional equivalents, and combinations thereof are bound to the modified ribosome or naturally occurring ribosome.

[0026] In some embodiments of the cell-free protein synthesis platform provided herein, one or more embedded membrane proteins comprises a membrane protein pore channel that allows passage of a polypeptide chain. In some embodiments, the ribosome can translocate a protein through the pore of the membrane protein. In some embodiments the membrane protein is a translocon. In some embodiments the membrane protein comprises a complex of proteins which comprise the translocon.

[0027] In some embodiments of the cell-free protein synthesis platform provided herein, the housing comprises a container with one or more partitions separating the container into two or more chambers comprising a reaction chamber and a product chamber. In some embodiments, the housing comprises a container with 1-3, 1-5, or 1-10 partitions separating the container into 2-4, 2-6, or 2-11 separate chambers. In some embodiments, the housing comprises the polymer membrane. In some embodiments, the one or more partitions comprise the polymer membrane. In some embodiments, the housing and components within the housing are comprised of apolymer. In some embodiments, the polymer material used to manufacture housing is selected from monomers and polymers that comprise the polymer membrane as described herein.

[0028] In some embodiments of the cell-free protein synthesis platform provided herein, the housing further comprises a membrane support structure. In some embodiments, the membrane support structure is a frame. In some embodiments, the frame is comprised of a polymer. In some embodiments, the housing comprises a vessel. In some embodiments, the vessel is manufactured form a polymer. In some embodiments the polymer is selected from the monomers and polymers listed herein. In some embodiments, the vessel contains a membrane support structure within.

[0029] In some embodiments of the cell-free protein synthesis platform provided herein, the housing comprises a flow reactor. In some embodiments, the flow reactor comprises one or more inlet / outlet connections. In some embodiments, the flow reactor comprises one or more inlet / outlet connections for product. In some embodiments, the flow reactor comprises the polymer membrane. In some embodiments, the polymer membrane of the flow reactor separates the flow reactor into one or more partitions. In some embodiments, the ribosome translates a protein through the polymer membrane pore into a product chamber of the flow reactor.

[0030] In some embodiments, the product chamber of the cell-free protein synthesis platform described herein further comprises a reaction chamber. In some embodiments, the reaction chamber comprises enzymes, substrates, and reagents. In some embodiments, enzymes assist in protein folding and / or, post-translational modification. In some embodiments reagents comprise macromolecular crowding reagents such as liposomes. In some embodiments protein folding enzymes include chaperone proteins. In some embodiments, the chaperone proteins are chaperonins. In some embodiments, chaperone proteins are selected from the group consisting of Chaperonin 10, Chaperonin 60, DnaK, FIMC, GroEL, GroES, GroESL, hchA, HSCP 70, HSP32, HSP70, HSPD1, PSMG2, PSMG4, SKP, functional equivalents, and combinations thereof. In some embodiments post-translational modification enzymes are selected from enzymes that perform the processes of acetylation, disulfide bond formation, glutathionylation, glycation, glycosylation, hydroxylation, lipidation, methylation, phosphorylation, proteolysis, S- nitrosylation, succinylation, SUMOylation, trisulfide bond formation, ubiquitination, functional equivalents, or combinations thereof. In some embodiments macromolecular crowding reagents are selected from the group consisting of Dextran 70, Ficoll 70, PEG, colloid-polymer mixtures, functional equivalents, and combinations thereof. In some embodiments, liposomes are preparedfrom cationic lipids, fatty acids, glycerolipids, ionizable lipids, PEGylated lipids, phospholipids, sterols, functional equivalents, or combinations thereof. In some embodiments, reaction chamber further comprises metal ions. In some embodiments metal ions are selected from the group consisting of Ca, Co, Cr, Cu, Fe, K, Mg, Mo, Mn, Na, Ni, V, Zn, functional equivalents, or combinations thereof.

[0031] In various embodiments, provided herein is a cell-free protein synthesis platform comprising: a ribosome bound to a polymer membrane comprising an affinity binding site and a polymer membrane pore, wherein the polymer membrane separates a reaction compartment from a product compartment. In some embodiments, the ribosome is bound to the polymer membrane by a membrane bound protein. In some embodiments, the membrane bound protein is a translocon. In some embodiments the membrane bound protein comprises a translocon and a hinge protein. In some embodiments, the platform comprises a hinge protein. In some embodiments, the membrane bound protein is a hinge protein.

[0032] In various embodiments provided herein is a kit for cell-free protein synthesis comprising a polymer membrane which comprises an affinity tag and a pore, and a large ribosome subunit attached to the polymer membrane. In some embodiments, the kit further comprises a reaction solution. In some embodiments, the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts. In some embodiments, the reaction solution comprises an excess of small ribosomal subunits. In various embodiments, the reaction solution comprises a great molar excess of small ribosomal subunits as compared with large ribosomal subunits. Large ribosomal subunits are, in various embodiments, bound, or attached to the polymer membrane and providing an excess of small ribosomal subunits as compared with large ribosomal subunits facilitates the efficiency of the platform and kits described in various embodiments herein. In some embodiments, the kit further comprising an mRNA for a protein of interest. Including an mRNA for a protein of interest is a way that the kits and platform provided herein can be tailored to individual user’s needs or to meet market demand. Different reaction solutions, ribosome, and vessel combinations described herein can be tailored to individual user’s needs or market demands.

[0033] In some embodiments, the kit comprises a dialysis bag comprising a polymer membrane with an affinity tag and a pore, and large ribosomal subunit attached to the polymer membrane. In some embodiments, the kit comprising the dialysis bag further comprises a reaction solution.In some embodiments, the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts. In various embodiments, the reaction solution with the kit comprising a dialysis bag comprises an excess of small ribosomal subunits. In various embodiments, there are a great excess of small ribosomal subunits as compared with large ribosomal subunits. In various embodiments, the kit comprising a dialysis bag comprises an mRNA for a protein of interest.

[0034] In various embodiments, the dialysis bag kit comprises mRNA for more than one protein of interest. In some embodiments the dialysis bag kit comprises mRNA for a variety of different proteins of interest. In various embodiments, the dialysis bag kit comprises mRNA for 1-10 proteins of interest. In various embodiments, the dialysis bag kit comprises an mRNA for 1-3, 1- 5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, 4-10, 1-25, 1-50, 1-100, 5-25, 5-50, 5-100, 25-50, 25-75, 25- 100 proteins of interest. In various embodiments, the reaction solution is inside the dialysis bag so that a protein gets made and translocated to the outside of the dialysis bag into the housing or vessel. In various embodiments, the reaction solution is outside of the dialysis bag in the housing or vessel so that a protein gets made within the bag and translocated to the inside of the dialysis bag.

[0035] In some embodiments, provided herein is a cell-free protein synthesis platform comprising: a large ribosome subunit, a polymer membrane comprising an affinity binding site and a polymer membrane pore, wherein the large ribosome subunit is bound to the polymer membrane and the polymer membrane separates a reaction compartment from a product compartment. In some embodiments, of the cell-free protein synthesis platform the large ribosome subunit is bound to the polymer membrane by a membrane bound protein. In some embodiments, the membrane bound protein is a translocon. In some embodiments, the membrane bound protein is a hinge protein. In some embodiments, the membrane bound protein is a translocon bound to a hinge protein. In some embodiments, the cell-free protein synthesis platform comprises a reaction solution. In some embodiments, the reaction solution comprises a cell-free protein extract. In some embodiments, the reaction solution comprises an excess of ribosome small subunits. In some embodiments, the cell-free protein synthesis platform further comprises a housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compartment. In some embodiments, the cell-free protein synthesis platform comprises a polymer membrane bag having an inside membrane surface and an outside membrane surface. In some embodiments, the affinity bindingsite is on the inside membrane surface of the polymer membrane bag. In some embodiments, the polymer membrane bag is a dialysis bag. In some embodiments, the large ribosome subunit ribosome is modified at one or more sites. In some embodiments, the large ribosome subunit is modified at one or more sites comprises and comprises 1 to 10 ribosome ligand binding sites.BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The following detailed description, given by way of example and not intended to limit the invention solely to the embodiments described herein, will best be understood in conjunction with the accompanying drawings, in which:

[0037] FIG. 1 is a diagram showing the interaction and components of the platform including a ribosome, the polymer membrane comprising affinity molecules, and their interaction as described herein. In FIG. 1 both of the ribosomal subunits are visible. The polymer membrane described herein is shown and the interaction between the ribosomal ligand binding site and the affinity molecules on the polymer membrane are shown as well. Also visible within the drawing is the polypeptide being produced by the platform through the polymer membrane pore.

[0038] ] FIG. 2 is a diagram showing the interaction and components of a ribosome with the polymer membrane tube, provided in various embodiments herein. In FIG. 2 both of the ribosomal subunits are visible. The polymer membrane tube described herein is shown and the interaction between the ribosomal ligand binding site and the affinity molecules on the polymer membrane tube are shown as well. Also visible within the drawing is a polypeptide being produced by the platform through the polymer membrane tube pore. Additionally, the reaction solution described in various embodiments comprises components labeled in FIG. 2, including Amino Acids, Buffers, Energy, Salts, and tRNA regenerating enzymes.

[0039] FIG. 3 is a diagram showing the interaction of a ribosomal ligand binding site with the polymer membrane containing affinity molecules. The diagram shows the chemical interaction between Ni affinity molecules and Histidine tag of the ribosomal ligand binding site.

[0040] FIG. 4 is a diagram showing the interaction and components of the platform including a ribosome, the polymer membrane comprising affinity molecules, and their interaction as described herein in various embodiments. In FIG. 4 both of the ribosomal subunits are visible. The polymer membrane described herein is shown and the interaction between the ribosomalligand binding site and the affinity molecules on the polymer membrane are shown. The affinity molecules on the polymer membrane shown in this diagram are antibodies. Also visible within the drawing is a polypeptide being produced by the platform through the polymer membrane pore.

[0041] FIG. 5 is a diagram showing the interaction and components the platform including a ribosome, the polymer membrane containing a membrane protein embedded within the polymer membrane, and their interaction as described herein in various embodiments. In FIG. 5 both of the ribosomal subunits are visible. Shown in the diagram is the interaction between the ribosomal ligand binding site and the membrane protein embedded within the polymer membrane. Also visible within the drawing is a polypeptide being produced by the platform through the embedded membrane protein pore.

[0042] FIG. 6. Pictured in the figure is an embodiment described herein comprising a dialysis bag. The diagram shows the polymer membrane as a dialysis bag. Additionally, a cap is shown with a hole for component addition. Pictured is also a holder for the cap to keep dialysis bag suspended within the solution of the container. FIG. 6 also shows a clamp used on the dialysis bag to seal off the bottom. All of this housed within the container and dashed lines indicate assembly.

[0043] FIG. 7. Pictured in the figure is an embodiment described herein comprising a dialysis bag. The diagram shows the components of an assembled dialysis bag CFPS reaction kit for small scale protein expression. The figure shows the cap with a hole attached to the dialysis bag supported by the cap holder keeping the assembly suspended within the container. The clamp is shown with dashed lines as it is inside of the container.

[0044] FIG. 8. Pictured in the figure is an embodiment described herein comprising a dialysis polymer membrane tube. The diagram shows the components of a polymer membrane tube CFPS reaction kit for small scale protein expression. Dashed lines indicate assembly. The figure shows a cap with a hole for component addition. Pictured is also a holder for the cap to keep the polymer membrane tube suspended within the solution of the container. The figure also shows the base inserted into the polymer membrane tube to seal off the bottom. All of this housed within the container and dashed lines indicate assembly.

[0045] FIG. 9. Pictured in the figure is an embodiment described herein comprising a dialysis bag. The diagram shows the components of a polymer membrane tube CFPS reaction kit forsmall scale protein expression. The figure shows a cap with a hole for component addition. Pictured is also a holder for the cap to keep the polymer membrane tube suspended within the solution of the container. The diagram shows the assembled polymer membrane tube inserted into the holder and left suspended in the container. The figure also shows the base with dashed lines inserted into the polymer membrane tube to seal off the bottom.

[0046] FIG. 10 shows an embodiment described herein comprising a kit. The diagram shows the components of a CFPS reaction kit for moderate scale protein expression. The polymer membrane is not shown in this diagram. The diagram shows the main housing with protruding grooves for the frame to slide into. The diagram shows the components of the frame, with one frame part containing holes and the other containing corresponding protrusions. The dashed lines indicate assembly.

[0047] FIG. 11 shows an embodiment described herein comprising a kit. The diagram shows the assembled components of a CFPS reaction kit for moderate scale protein expression. The polymer membrane is not shown in this diagram. The diagram shows the main housing assembled by combining frame halves and sliding into protruding grooves. The diagram shows the assembled components of the frame, with one frame part containing holes and the other containing corresponding protrusions.

[0048] FIG. 12 shows an embodiment described herein comprising a kit. Pictured are components of a CFPS reaction kit for moderate scale protein expression. The polymer membrane is shown in this diagram. The diagram shows the main housing with a recessed groove for the frame assembly to slide into. The diagram shows the components of the frame, with one frame pail containing holes and the other containing corresponding protrusions. Also shown is the polymer membrane with holes corresponding with the frame components. The dashed lines indicate assembly.

[0049] FIG. 13 shows an embodiment described herein comprising a kit. Pictured are components of a CFPS reaction kit for moderate scale protein expression. The polymer membrane is shown in this diagram. The diagram shows the main housing with a recessed groove that the frame containing the membrane has been slid into. The diagram shows the components of the frame, with one frame part containing holes and the other containing corresponding protrusions and the polymer membrane in between.

[0050] FIG. 14 shows an embodiment described herein comprising a flow reactor. Pictured arecomponents of a CFPS flow reactor for large scale protein expression. The diagram shows the components of the flow reactor. Pictured are both chamber caps of the reactor containing a process connection. Also shown are the perforated base plates. The chamber of the flow reactor is shown with both process connections. The polymer membrane tubes are shown as a group. The dashed lines indicate assembly of the components.

[0051] FIG. 15 shows an embodiment described herein comprising a flow reactor. Pictured are components of a CFPS flow reactor for large scale protein expression. The diagram shows the assembled components of the flow reactor. Pictured are both chamber caps of the reactor containing a process connection. Also shown are the perforated base plates. The chamber of the flow reactor is shown with both process connections. The polymer membrane tubes are shown as a group inserted into holes in both the top and bottom perforated base plates. The chamber caps with process connections are shown attached to the top and bottom perforated base plates and chamber.

[0052] FIG. 16 shows the Candida albicans 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L26B, L31B, L35A are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0053] ] FIG. 17 shows the Danio rerio 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L31 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0054] ] FIG 18 shows the Drosophila melanogaster 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L26, L31, L35 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0055] FIG. 19 shows the Escherichia coli 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22, L23, L24, L29 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0056] FIG. 20 shows the Haloarcula marismortui 50S ribosomal subunit structure from theoutside perspective. Ribosomal proteins L22P, L23P, L24P, L29P, L31e are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0057] FIG. 21 shows the Homo sapiens 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L26, L31, L35 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0058] FIG. 22 shows the Kluyveromyces lactis 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L25, KLLA0A06336p, KLLA0B02937p, KLLA0B05742p, KLLA0E12453p, KLLA0F05247p are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0059] FIG. 23 shows the Mus musculus 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L19, L26, L31, L35 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0060] FIG. 24 shows the Nicotiana tabacum 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22, L22-2, L23a, L26, L31, L35, L38 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0061] FIG. 25 shows the Oryctolagus cuniculus 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L24, L29, SRP19, SRP54, are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0062] FIG. 26 shows the Pseudomonas aeruginosa 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L23, L24, L29, L32 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and 1labeled.

[0063] FIG. 27 shows the Pyrococcus furiosus 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L3P, L24P, L29P, L31e are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0064] FIG. 28 shows the Saccharomyces cerevisiae 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L23, L24, L29 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0065] FIG. 29 shows the Spinacia oleracea 50S Chloroplast ribosomal subunit structure from the outside perspective. Ribosomal proteins rpLI7, rpL22, rpL26, rpL31, rpL38 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0066] FIG. 30 shows the Staphylococcus aureus 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22, L23, L24, L29 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0067] FIG. 31 shows the Sus scrofa 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins eL22, uL22, L23, L24, L29, L31, L38 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.

[0068] FIG. 32 shows the Thermits thermophilus 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L23, L24, L29, L32 are highlighted. These ribosomal proteins may be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Ribosome tunnel opening is also visible and labeled.DETAILED DESCRIPTION

[0069] The inventor has recognized a need for advances in cell-free protein synthesis that provide scalability and simplicity in the production of proteins of interest. Such advances will be particularly valuable, for example, in synthetic biology applications.

[0070] Provided herein, in various embodiments, is cell-free protein synthesis platform, comprising a ribosome and a polymer membrane comprising a polymer membrane pore and an affinity binding site capable of binding the ribosome, wherein the polymer membrane separates a reaction compartment from a product compartment. In some embodiments, the ribosome is bound to the polymer membrane. In some embodiments, the ribosome is bound to the polymer membrane by a membrane bound protein. In some embodiments, the membrane bound protein is a translocon. In some embodiments, the membrane bound protein is a hinge protein. In some embodiments, the membrane bound protein is a translocon bound to a hinge protein. In some embodiments, the cell-free protein synthesis platform further comprises a reaction solution. In some embodiments the reaction solution comprises a cell-free protein extract. In some embodiments, the cell-free protein extract comprises an excess of small ribosomal subunits. In some embodiments, the cell-free protein synthesis platform further comprises a housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compartment. In some embodiments, the cell-free protein synthesis platform further comprises a polymer membrane bag having an inside membrane surface and an outside membrane surface. In some embodiments, the affinity binding site is on the inside membrane surface of the polymer membrane bag. In some embodiments, the polymer membrane bag is a dialysis bag. In some embodiments, the ribosome is a modified ribosome. In some embodiment, the modified ribosome comprises one or more modified ribosome sites, and in some embodiments, the modified ribosome comprises 1 to 10 ribosome ligand binding sites.

[0071] In some embodiments of the cell-free protein synthesis platform provided herein, the modified ribosome comprises a ribosome ligand binding site, in some embodiments, the modified ribosome comprises two or more ribosome ligand binding sites, in some embodiments, the ribosome ligand binding sites are different ligand binding sites. In some embodiments, the ribosome ligand binding site is a molecular binding site. In some embodiments, the ribosomeligand binding site is an antibody epitope, in some embodiments, the ribosome ligand binding site is selected from His-tag, His-Glu-tag, Asp-Lys-tag, Glu-tag, Arg-tag, Cys-tag, FLAG-tag, Capture Select C-tag, TC-tag, Xpress-tag, Softag-3-tag, Strep-tag, Aul, AU5, T7-tag, V5-tag, B- tag, E2 epitope, HSV epitope, KT3 epitope, Myc epitope, PDZ ligands, Asp-tag, Phe-tag, Protein C, Sl-tag, Universal-tag, VSV-G, a HTTPHH, functional equivalents, or combinations thereof. In some embodiments the ribosome ligand binding site is on a ribosomal protein. In some embodiments, the ribosomal protein comprising the ribosome ligand binding site is from a different organism than the ribosome. In some embodiments, the ribosomal protein is modified from its native form. In some embodiments, the ribosome ligand binding site is on a ribosomal RNA. In some embodiments, the ribosomal RNA is from a different organism than the ribosome. In some embodiments, the ribosomal RNA is modified from its native form.

[0072] In some embodiments of the cell-free protein synthesis platform provided herein, the affinity binding site comprises an affinity molecule. In some embodiments, the affinity molecule is capable of binding a ribosome ligand binding site. In some embodiment, the affinity binding site comprises an affinity molecule that binds a ribosome ligand binding site.

[0073] In some embodiments of the cell-free protein synthesis platform provided herein the ribosome is attached to the polymer membrane. In some embodiments, the ribosome is bound to one or more affinity molecules on the polymer membrane. In some embodiments, the ribosome is bound to 1 to 10 affinity molecules on the polymer membrane. In some embodiments, the ribosome contains a naturally occurring ribosome ligand binding site. In various embodiments, the ribosome ligand binding site is capable of binding an affinity molecule described herein.

[0074] In some embodiments of the cell-free protein synthesis platform provided herein the affinity molecule is bound near the polymer membrane pore. In some embodiments, the affinity molecule is bound on the polymer membrane pore edge. In various embodiments, the affinity molecule is 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5- 50 nm, 0.5-25 nm, 0.5 nm-10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5- 100 nm, 5-50 nm, 5-25 nm, or 5-10 nm from the polymer membrane pore edge. In some embodiments, the affinity molecule is selected from a metal chelating tag, an antibody epitope, a covalent binding molecule, and functional equivalents and combinations thereof. In some embodiments, the affinity molecule is selected from HAT-tag, His-tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1, AU5, B-tag, E2,EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl- Sepharose (Phe-tag), ProC,SplAsH (Cys-tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA-aptamer, RNA-Sephadex, RNA-Streptavadin), Tags made by the SELEX process, or combinations thereof.

[0075] In some embodiments of the cell-free protein synthesis platform provided herein each polymer membrane pore has 1 to 10 affinity binding molecules within 100 nm of the edge of the polymer membrane pore.

[0076] In some embodiments of the cell-free protein synthesis platform provided herein the ribosome is a reconstituted ribosome, a recombinant ribosome, or a hybrid ribosome.

[0077] In some embodiments of the cell-free protein synthesis platform provided herein the ribosome ligand binding site is attached via one or more inteimediate molecules to the affinity molecule attached to the polymer membrane. In some embodiments, the intermediate molecule is a signal recognition protein. In some embodiments, the signal recognition protein is selected from SEQ ID NO: 257 to SEQ ID NO: 271, functional equivalents, or combinations thereof. In some embodiments, the intermediate molecule is a polymer, antibody, antibody complex, small molecule chemical linker, dendrimer, hinge protein, or combinations thereof. In some embodiments, the ribosome ligand binding site is attached via 2-10 intermediate molecules to the affinity molecule on the polymer membrane.

[0078] In some embodiments of the cell-free protein synthesis platform provided herein comprises a hybrid ribosome comprises molecules selected from two or more organisms. In some embodiments, the hybrid ribosome comprises molecules selected from archaea, eukaryotes, prokaryotes, or combinations thereof.

[0079] In some embodiments the cell-free protein synthesis platform provided herein a modified ribosome protein. In some embodiments, the modified ribosome protein comprises modifications selected from amino acid substitution, protein tags, non-canonical amino acid substitution, or combinations thereof. In some embodiments, the modified ribosome protein is selected from SEQ ID NO:1 to 256, SEQ ID NO: 272 to SEQ ID NO:490, functional equivalents, or combinations thereof.

[0080] In some embodiments of the cell-free protein synthesis platform provided herein, the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts. Insome embodiments, the reaction solution further comprises an excess of small ribosomal subunits. In some embodiments, the reaction solution further comprises an mRNA for a protein of interest. In some embodiments, the cofactors in the cell free protein synthesis extract comprise proteins, ions, and RNA. In some embodiments, the tRNA’s comprise naturally occurring, modified and synthesized tRNA molecules. In some embodiments, amino acids are selected from naturally occurring amino acids, noncanonical amino acids, or combinations thereof. In some embodiments, the amino acids are selected from is selected from alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine, pryrrolysine (stop codon amino acids), homoallylglycine, homopropargylglycine, azidohomoalanine, azidonorleucine, azidophenylalanine, acetylphenylalanine, acetylphenylalanine, propargyloxyphenylalanine, functional equivalents, or combinations thereof. In some embodiments, the energy molecules comprise nucleoside triphosphates (NTPs).

[0081] In some embodiments of the cell-free protein synthesis platform provided herein, the polymer membrane is comprised of a polymer which comprises monomers or polymers selected from cellulose acetate, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polyester, polyether sulfone, polyetherether ketone, polyetherimide, polyethersulfone, polyethylene, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfone, polytetrafluoroethylene, polyvinyl chloride, polyvinylidene fluoride, polyamide, polyacrylonitrile, polyethersulfone, polyvinylidene fluoride, functional equivalents, or combinations thereof. In some embodiments, the polymer membrane is a hydrogel. In some embodiments, the polymer membrane has a thickness of at least 1 nm, 5 nm, 1 Onm, 15 nm, 20 nm, 25 nm, 30 nm, 50 nm, 75 nm, or 100 nm. In some embodiments, the polymer membrane has a thickness selected from of 1-100 nm, 1-75 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-75 nm, 5-50 nm, 5-25 nm, 5-10 nm, 10-100 nm, 10-75 nm, 10-50 nm, 10-25 nm, 25-100 nm, 25-75nm, or 25-50nm. In some embodiments, the polymer membrane has a thickness of 5 nm to 10 nm. In some embodiments, the polymer membrane is comprised of one or more layers. In some embodiments, the polymer membrane is comprised of 1-2, 1-3, 1-5, 1- 10, 2-5, 2-10, or 5-10 layers. In some embodiments the polymer membrane comprises one or more layers of a polymer support matrix. In some embodiments, the polymer membrane is asemi permeable membrane. In some embodiments, the polymer membrane is selectively permeable to water and ions of interest. In some embodiments, the polymer membrane pores have a diameter of about 3 nm to about 15 nm. In some embodiments, the polymer membrane pores have a diameter selected from 5-100 nm, 5-75 nm, 5-50 nm, 5-25 nm, 5-10 nm, 10-10 Onm, 10-75 nm, 10-50 nm, 10-25 nm, 20-100 nm, 20-75 nm, 20-50 nm, 20-25 nm, 50-100 nm, or 50-75 nm. In some embodiments, the polymer membrane pores are spaced at least 10 nm, 20 nm, 25 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, or 500 nm edge to edge apart from other membrane pores. In some embodiments the polymer membrane comprises 1-10 affinity molecules within 10 nm of the edge of each membrane pore. In some embodiments, the polymer membrane comprises 1-5 affinity molecules within 10 nm of the edge of each membrane pore. In some embodiments, the polymer membrane comprises one or more coatings. In some embodiments, the polymer membrane comprises one or more embedded membrane protein. In some embodiments, the embedded membrane protein is a translocon. In some embodiments, the translocon binds to a modified ribosome. In some embodiments, the translocon binds a hinge protein, intermediate molecule, dendrimer, antibody, antibody complex, or combinations thereof. In some embodiments, a hinge protein, intermediate molecule, dendrimer, antibody, antibody complex, or combinations thereof binds to the modified ribosome. In some embodiments, the one or more embedded membrane proteins comprises a membrane protein pore channel that allows passage of a polypeptide chain.

[0082] In some embodiments of the cell-free protein synthesis platform provided herein, the ribosome can translocate a protein through the pore of the membrane protein.

[0083] In some embodiments of the cell-free protein synthesis platform provided herein, the housing comprises a container with one or more partitions separating the container into two or more chambers comprising a reaction chamber and a product chamber. In some embodiments, the housing comprises a container with 1-3, 1-5, or 1-10 partitions separating the container into 2-4, 2-6, or 2-11 separate chambers. In some embodiments, the housing comprises the polymer membrane. In some embodiments, the one or more partitions comprise the polymer membrane.

[0084] In some embodiments of the cell-free protein synthesis platform provided herein, the housing comprises a vessel. In some embodiments, the vessel is comprised of a polymer comprised of monomers or polymers selected from acrylamide, 3-acrylamidopropyl trimethylammonium chloride, N-hydroxymethyl acrylamide. N,N-methylenebisacrylamide,cellulose acetate, N,N-dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n-butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, poly sacrylamido-2-methyl- 1 -propane- sulfonic acid, polyacrylic acid, polymethacrylic acid, polyisopropylacrylamide, poly-3- acrilamidopropyl trimethylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polydiallyldimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide, isopropylacrylamide, or vinylpyrrolidonc. copolymers of acrylamide-2-methyl- 1 propanesulfonic acid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido- propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl- pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidone, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfide, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl- ammonium, polyvinyl chloride, polyvinylidene fluoride, poly-4- vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinylsulfonic acid, functional equivalents and combinations thereof. In some embodiments, the vessel contains a membrane support structure.

[0085] In some embodiments of the cell-free protein synthesis platform provided herein comprising a housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compartment, the housing comprises a flow reactor. In some embodiments, the flow reactor comprises one or more inlet / outlet connections. In some embodiments, the flow reactor comprises one or more inlet / outlet connections for product. In some embodiments, the flow reactor comprises the polymer membrane. In some embodiments, the polymer membrane of the flow reactor separates the flow reactor into one or more partitions. In some embodiments, the ribosome translates a protein through the polymer membrane pore into a product reaction chamber of the flow reactor. The polymer membrane can, in various embodiments, be a polymer as described herein in various embodiments.

[0086] In some embodiments, provided herein is a kit for cell-free protein synthesis comprising a polymer membrane which comprises an affinity tag and a pore, and a large ribosome subunit attached to the polymer membrane. In some embodiments, the kit further comprises a reaction solution. In some embodiments, the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts. In some embodiments, the reaction solution comprises an excess of small ribosomal subunits. In some embodiments, the kit further comprises an mRNA for a protein of interest.

[0087] In some embodiments, provided herein is a kit for cell-free protein synthesis comprising a dialysis bag which comprises a polymer membrane with an affinity tag and a pore, and large ribosomal subunit attached to the polymer membrane. In some embodiments, the kit further comprises a reaction solution. In some embodiments, the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts. In some embodiments, the reaction solution comprises an excess of small ribosomal subunits. In some embodiments, the kit for cell- free protein synthesis comprising a dialysis bag which comprises a polymer membrane with an affinity tag and a pore, and large ribosomal subunit attached to the polymer membrane further comprises an mRNA for a protein of interest. In some embodiments, the reaction solution is inside the dialysis bag.

[0088] In some embodiments the cell-free protein synthesis platform provided herein, further comprises a reaction solution. In some embodiments, the reaction solution comprises a cell-free protein synthesis extract.

[0089] In some embodiments of the cell-free protein synthesis platform provided herein the reaction solution comprises an excess of ribosome small subunits.

[0090] In some embodiments the cell-free protein synthesis platform provided herein, further comprises housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compailment.

[0091] In some embodiments the cell-free protein synthesis platform provided herein further comprises dendrimers that can be used for controlling pore diameter and distribution. In some embodiments, the dendrimers contain affinity molecules. In some embodiments, the dendrimer affinity molecules are selected form HAT-tag, His-tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1, AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin bindingpeptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl-Sepharose (Phe-tag), ProC, SplAsH (Cys-tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA- aptamer, RNA-Sephadex, RNA-Streptavadin), tags made by the SELEX process, functional equivalents or combinations thereof. In some embodiment, the dendrimer material is selected from carbosilane, citric acid, glycodendrimers, peptide, phosphorus, polyamido-amine, polyether, polyglycerol, poly L-lysine, polypropylene-imine, triazine, functional equivalents or combinations thereof.

[0092] In some embodiments of the cell-free protein synthesis platform provided herein, the product compartment further comprises a reaction chamber. In some embodiments, the reaction chamber further comprises enzymes, substrates, and reagents. In some embodiments, the enzyme are enzymes that assist in protein folding, post-translational modification, or combinations thereof. In some embodiments, the enzymes that assist in protein folding are chaperone proteins. In some embodiments, the chaperone proteins are chaperonins. In some embodiments, the enzymes are selected from Chaperonin 10, Chaperonin 60, DnaK, FIMC, GroEL, GroES, GroESL, hchA, HSCP 70, HSP32, HSP70, HSPD1, PSMG2, PSMG4, SKP, functional equivalents, and combinations thereof. In some embodiments, the enzymes that perform post- translational modification are enzymes that catalyze acetylation, disulfide bond formation, glutathionylation, glycation, glycosylation, hydroxylation, lipidation, methylation, phosphorylation, proteolysis, S-nitrosylation, succinylation, SUMOylation, trisulfide bond formation, ubiquitination, functional equivalents, and combinations thereof.

[0093] In some embodiments, the ribosome utilized within the platform may be a modified ribosome. This ribosome is modified to contain affinity molecule ligand binding sites on one or more of the ribosomal proteins. In some embodiments, these modifications arc selected from Table 1. In some embodiments, these modifications are a combination selected from Table 1. In some embodiments combinations of ligand binding sites are used to orient the ribosome over the pore to ensure polypeptide is translated through the polymer membrane pore and not into the reaction compartment. In some embodiments the ribosome ligand bindings sites are selected from HAT-tag, His-tag, anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1, AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl-Sepharose (Phe-tag), ProC,SplAsH (Cys-tag), functional equivalents, andcombinations thereof. Any protein purification methods and the affinity molecules associated with them, either currently available, or developed in the future, can be, as understood by persons of skill in the art, used with the cell-free protein synthesis platform described herein in various embodiments. As is understood by persons of skill in the art, any affinity molecule suitable for protein purification can be used in the platform described herein in various embodiments.

[0094] In some embodiments, the ribosome is modified to contain a RNA ligand binding site on the ribosomal RNA. These can in various embodiments bind RNA-affinity tags. In some embodiments ribosomal RNA modifications can be RNA-affinity tags selected from RNA-biotin, RNA-aptamer, RNA-Sephadex, RNA-Streptavadin, functional equivalents, and combinations thereof. SELEX (Systematic evolution of ligands by exponential enrichment) is a method that can be used to develop new RNA affinity tags. RNA affinity molecules on or near the polymer membrane pore bind pore bind ligand binding sites on the ribosomal RNA. RNA affinity tags are an ongoing field of study. Persons of skill in the art understand that any suitable RNA affinity tag can be used with the platform described herein in various embodiments.

[0095] In some embodiments, larger intermediate molecules such as dendrimers with affinity components, antibodies or antibody complexes (ex. ELISA / ELISA sandwich), signal recognition peptides, dendrimers, or other functionally equivalent molecules may be used to bind to ligand sites on the ribosome and affinity molecules on the polymer membrane.

[0096] In some embodiments, the ribosome is modified so the ribosomal subunits are tethered together by a linker. In some embodiments, the linker is a polymer with affinity molecules. In some embodiments, the polymer linker attaches to modified ribosomal proteins on the large and small ribosomal subunits. In some embodiments, the polymer linker comprises a polymer chain or structure, and affinity molecules. In some embodiments, the polymer material comprises a monomer or polymer selected from acrylamide, 3-acrylamidopropyl trimethylammonium chloride, N-hydroxy methyl acrylamide, N,N-methylenebisacrylamide, cellulose acetate, N,N- dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n- butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, polysacrylamido-2-methyl-l -propane-sulfonic acid, polyacrylic acid, polymethacrylic acid, polyisopropylacrylamide, poly-3-acrilamidopropyl trimethylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole,T1polycarbonate, polydially Idimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide, isopropylacrylamide, or vinylpyrrolidonc. copolymers of acrylamide-2-methyl-lpropanesulfonic acid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido-propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl-pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidone, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfone, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl-ammonium, polyvinyl chloride, polyvinylidene fluoride, poly-4-vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinyl sulfonic acid, functional equivalents, or combinations thereof. In some embodiments, the more commonly used industrial polymers, polyamide, polyacrylonitrile, poly ether sulfone, polyvinylidene fluoride, functional equivalents, or combinations thereof.

[0097] In some embodiments the polymer linker affinity molecule is selected from HAT-tag, His- tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1, AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl-Sepharose (Phe-tag), ProC, SplAsH (Cys-tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA-aptamer, RNA-Sephadex, RNA-Streptavadin), tags made by the SELEX process, functional equivalents, and combinations thereof. In some embodiments, the polymer linker is a dendrimer. In some embodiments the dendrimer material is selected from carbosilane, citric acid, glycodendrimers, peptide, phosphorus, polyamido-amine, polyether, polyglycerol, poly L-lysine, polypropylene-imine, triazine, functional equivalents, or combinations thereof.

[0098] In various embodiments described herein, the ribosomal ligand binding sites are attached to affinity molecules on the polymer membrane on or near the pore edge. In some embodiments, combinations of ribosomal protein and ribosomal RNA ligand binding sites are used to orient the ribosome so that the polypeptide being translated out of the ribosome exit tunnel is translocatedthrough the polymer membrane pore to the product compailment.

[0099] In some embodiments, the affinity molecules are located on the edge of the membrane pore. In some embodiments each polymer membrane pore has 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3- 10, 4-6, or 4-10 affinity binding molecules. In some embodiments, the affinity molecules are located within 10 nm of the edge of the membrane pore. In some embodiments, the affinity molecule is 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5- 50 nm, 0.5-25 nm, 0.5-10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-50 nm, 5-25 nm, or 5-10 nm from the polymer membrane pore edge. In some embodiments each polymer membrane pore has 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 affinity binding molecules within 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5-50 nm, 0.5-25 nm, 0.5-10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-50 nm, 5-25 nm, or 5-10 nm from the polymer membrane pore edge. In some embodiments, this attachment aligns the ribosome tunnel opening over the polymer membrane pore so that the expressed polypeptide is translated through the polymer membrane pore into the product compartment of the platform. In some embodiments, having multiple affinity molecules allows for a stronger attachment of the ribosome to the polymer membrane. In some embodiments, multiple affinity molecules and combinations allow for control over orientation and binding of the ribosome to the polymer membrane.

[0100] In some embodiments, the polymer membrane pore dimensions, height and diameter, can be varied to allow for variability in the environment for the protein to fold. In some embodiments, the polymer membrane pore dimensions, height and diameter are determined in advance by the user in accordance with the needs of the protein production process. In some embodiments, there is flexibility, or variability, in the polymer membrane pore dimensions, height and diameter, that can account for the protein folding process. Methods and systems that can be used to control pore dimensions are understood by persons of skill in the ait who will, in various embodiments, tailor the polymer membrane pores in accordance with the protein of interest. For example, the protein folding process for a protein under certain conditions can be considered by persons of skill in the ait when considering appropriate pore dimensions and flexibility.

[0101] In some embodiments, manufacturing methods are used to control pore dimension and distribution within the polymer membrane. In some embodiments pore dimensions anddispersion are controlled by using dendrimers within the polymer membrane during manufacturing. In some embodiments the dendrimer material is selected from carbosilane, citric acid, glycodendrimers, peptide, phosphorus, polyamido-amine, polyether, polyglycerol, poly L- lysine, polypropylene-imine, sulfonimide, triazine, functional equivalents, or combinations thereof. In some embodiments the dendrimer further comprises affinity molecules. In some embodiments, the affinity molecule is selected from HAT-tag, His-tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1, AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl- Sepharose (Phe-tag), ProC,SplAsH (Cys-tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA-aptamer, RNA-Sephadex, RNA-Streptavadin, tags made by the SELEX process, functional equivalents and combinations thereof. In some embodiments, a larger pore could allow for larger proteins to self-assemble. In some embodiments the polymer membrane pores are spaced apart at variable distances, so that individual ribosomes do not chemically interfere with nearby ribosomes during translation and to allow for greater access of reagents to the translating ribosome. In some embodiments the polymer membrane is comprised of several layers. In these embodiments, the polymer membrane can be designed to mimic the hydrophilic / hydrophobic regions of a naturally occurring phospholipid bilayer membrane by using multiple layers of different polymers.

[0102] In some embodiments, the polymer membrane comprises a material of a monomers or polymers selected from the group consisting of acrylamide, 3-acrylamidopropyl trimethylammonium chloride, N-hydroxymethyl acrylamide, N,N-methylenebisacrylamide, cellulose acetate, N,N-dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n-butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, polysacrylamido-2-methyl-l-propane- sulfonic acid, polyacrylic acid, polymethacrylic acid, polyisopropylacrylamide, poly-3- acrilamidopropyl trimethylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polydiallyldimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide,isopropylacrylamide, or vinylpyrrolidone, copolymers of acrylamide-2-methyl-lpropanesulfonic acid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido- propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl- pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidone, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfone, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl- ammonium, polyvinyl chloride, polyvinylidene fluoride, poly-4-vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinyl sulfonic acid, functional equivalents, and combinations thereof.

[0103] In some embodiments, readily available industrial polymers such as polyamide, polyacrylonitrile, polyether sulfone, polyvinylidene fluoride can be used to make the membrane. In some embodiments the polymer membrane is manufactured using methods known to persons of skill in the art, such as step-growth polymerization (formed through a series of individual steps / reactions, include condensation reactions, include polyesters, polyamides etc.), chaingrowth polymerization (steps include initiation, termination, propagation, chain transfer) and controlled polymerization (methods include anionic / cationic polymerization, block copolymers, end-functional polymers, branched polymers, ring-opening polymerization) and affinity molecules attached using similar methods. In some embodiments, the membrane can be manufactured by methods such as casting, electrospinning, interfacial polymerization, phase inversion, spraying, and combinations thereof known to those skilled in the art. In some embodiments these methods and materials can be used to manufacture the polymer membrane into tubes, suitable for certain embodiments such as a flow reactor or tube embodiment. In some embodiments these methods can be used to manufacture a support structure for the polymer membrane using polymers listed above.

[0104] In some embodiments dendrimers embedded within the polymer membrane during manufacturing can be used to control pore dimensions and dispersion via centrifugation. These dendrimers can, in some embodiments, be later removed via using an appropriate solvent. In some embodiments, laser etching can be used to manufacture pores within the polymer membrane. In some embodiments the dendrimer material is selected from carbosilane, citric acid, glycodendrimers, peptide, phosphorus, polyamido-amine, polyether, polyglycerol, poly L-lysine, polypropylene-imine, sulfonimide, triazine, functional equivalents, or combinations thereof.

[0105] In some embodiments, laser etching can be used to manufacture pores within the polymer membrane. In some embodiments pores are further reacted with affinity molecules to incorporate the base structure of the affinity molecules into the polymer membrane edge. The base structure comprises a variable polymer chain.

[0106] In some embodiments a membrane protein can be embedded within the polymer membrane structure. In some embodiments, kits to capture membrane proteins can be used to embed membrane proteins within the polymer membrane structure. An example of such a kit is the MembraneMax™ sold by Invitrogen. In these embodiments the membrane protein can be a translocon and would enable ribosomes to be attached either directly or through an intermediate molecule such as a hinge protein, signal recognition peptide, or a dendrimer. In these embodiments this would allow for the cell-free protein synthesis platform to more closely resemble a naturally occurring ribosome nascent complex in association with a translocon.

[0107] In some embodiments the polymer membrane can have a coating. In these embodiments the coating can prevent agglutination of both proteins in the reaction compartment and in the product compartment. In some embodiments, the coating is comprised of naturally occurring polymers such as cellulose. The coating can be attached to the polymer membrane postmanufacturing. Those skilled in the art can select, in various embodiments, appropriate coatings depending on chemical and protein synthesis environmental conditions best suited for different embodiments and proteins of interest. Coating can comprise monomers and polymers listed herein.

[0108] In some embodiments the reaction solution contains a commercially available cell-free extract. In various embodiments, commercially available cell-free protein synthesis systems, reactions, or kits are selected from Arbor Biosciences kits (myTXTL), Bioneer kits (AccuRapid), Biotech Rabbit kits (RTS 100, RTS 500, RTS 500 ProteoMaster, RTS 9000, RTS 100 Wheat Germ, RTS 500 Wheat Germ, RTS 100 Insect), Cayman Chemical kits, Creative Biomart kits, Dispendix kits (I.DOT), GenScript kits (TurboCHO), LenioBio kits (ALiCE), New England Biolabs kits (PUR Express, NEB Express), Promega kits (Flexi), SinoBiological kits, Takara kits, ThermoFisher kits, and the like, or combinations thereof. In some embodiments the reaction solution comprises a cell-free extract prepared by the manufacturer of the platform. In someembodiments, the reaction solution comprises a cell-free extract prepared by or the end user. The methods to prepare these extracts are well documented in literature and are understood by persons of skill in the art. In some embodiments, some reaction solution components can be prepared using various embodiments of the platform described herein. In some embodiments, the reaction solution contains a high ration of smaller ribosomal subunits. In these embodiments, the high ratio of smaller ribosome subunits increases the availability of the subunits that can bind the larger ribosome subunits that are attached to the membrane. This would increase the likelihood of these subunits’ association and initiating translation, thereby increasing the yield and efficiency of the platform.

[0109] In some embodiments tRNA molecules can be added by the end user. In some embodiments, engineered tRNA’s can be added to the reaction mixture. In some embodiments noncanonical amino acids can be added to the reaction solution. Noncanonical amino acids used in various embodiments can include, for example, homoallylglycine, homopropargylglycine, azidohomoalanine, azidonorleucine, azidophenylalanine, acetylphenyl-alanine, acetylphenylalanine, propargyloxyphenylalanine, functional equivalents and combinations thereof.

[0110] In some embodiments, tRNA aminoacyl-synthetases can be added to the reaction solution. In some embodiments, tRNA aminoacyl synthetases used to incorporate non-canonical amino acids can be added to reaction mixture by platform manufacturer or end user. Addition of these enzymes, in various embodiments, recycles used tRNA molecules and allows for increased yield and efficiency of the platform. In some embodiments the energy molecules and / or energy systems included in the reaction solution comprise creatine phosphate / creatine kinase, glucose, glutamate decarboxylase, maltodextrin, nucleoside triphosphates (NTPs), phosphoenolpyruvate, 3-phosphoglycerate, functional equivalents, and combinations thereof. In some embodiments noncanonical nucleotides can be included in the reaction solution.

[0111] In some embodiments the reaction solution contains one or more buffers. Buffers that can be used, in various embodiments, include, for example, MES, Bis-Tris, ADA, ACES, PIPES, MOPSO, Bis-Tris Propane, BES, MOPS, TES, HEPES, DIPSO, MOBS, TAPSO, Tris or Trizma®, HEPPSO, POPSO, TEA, EPPS, Tricine, Gly-Gly, Bicine, HEPBS, TAPS, AMPD, TABS, AMPSO, CHES, CAPSO, AMP, CAPS, CABS, other functionally equivalent buffer system, and combinations thereof. In some embodiments buffers are added by a platformmanufacturer. In some embodiments buffers are added by an end user.

[0112] In some embodiments, mRNA coding for the protein of interest is added by the end user. In some embodiments, kits or platforms can be provided along with mRNA coding for a protein of interest. In some embodiments the reaction solution contains enzymes to process DNA to RNA to mRNA. In some embodiments these enzymes can be sold with the platform. In some embodiments, commercially available mRNA synthesis kits are used to make mRNA. In some embodiments, commercially available mRNA synthesis kits can be used. In various embodiments, commercially available mRNA synthesis kits can be selected from ApexBio (mCAP, HyperScribe™), Creative Biogene (RNTK-001, RNTK-002), New England Biolabs (HiScribe® with CleanCap®), System Biosciences (mRNAExpress™), Takara (IVTpro™), ThermoFisher (MegaScript™, mMESSAGE mMACHINE™), functional equivalents, or combinations thereof.

[0113] In some embodiments, the platform housing comprising the platform resembles a dialysis bag. In some embodiments, the platform housing is a dialysis bag. In these embodiments, the dialysis bag provides a cost-effective version of the platform and can be variable in size and used for a variety of applications. It is envisioned by the inventor that users can easily tailor the dialysis bag embodiments of the platform and quickly and flexibly use it. In some embodiments, the ribosomes are bound to the dialysis bag membrane from inside of the housing. In various embodiments, the protein synthesis process translates the protein through the polymer membrane of the bag into the external containment vessel. In some embodiments the ribosomes are bound to the outside of the dialysis bag membrane. In these embodiments the protein is produced outside of the dialysis bag and the protein is translated through the polymer membrane of the dialysis bag into the internal compartment of the dialysis bag. These embodiments provide a wide range of control over the chemical environment of the platform and allow for high purity of the proteins being translated.

[0114] In some embodiments the product chamber further comprises a reaction chamber. In some embodiments the reaction chamber comprises enzymes, one or more substrates, and reagents. In some embodiments enzymes assist in protein folding and / or post-translational modification. In some embodiments reagents comprise macromolecular crowding reagents such as liposomes. In some embodiments protein folding enzymes include chaperone proteins such as chaperonins. In some embodiments enzymes that assist in protein folding and / or post-translational modificationare selected from those describe herein, in various embodiments. In some embodiments post- translational modification enzymes can be selected from those described herein in various embodiments. In some embodiments, liposomes are prepared from cationic lipids, fatty acids, glycerolipids, ionizable lipids, PEGylated lipids, phospholipids, sterols, functional equivalents, and combinations thereof. In some embodiments, reaction chamber further comprises metal ions. In some embodiments metal ions are selected from Ca, Co, Cr, Cu, Fe, K, Mg, Mo, Mn, Na, Ni, V, Zn, functional equivalents, and combinations thereof.

[0115] The cell-free protein synthesis platform described herein in various embodiments provides high yields and fast delivery of proteins of interest. Target yields of protein for the platform described herein in various embodiments are around 1 g / L of reaction volume. The cell- free protein synthesis platform, in various embodiments, can generate up to 100 mg of protein within 24-72 hours, depending on reaction volume and membrane surface area. In some embodiments, the target yield of protein is 1-100 g / L, 1-75 g / L, 1-50 g / L, 1-25 g / L, 1- 10 g / L, 1-5 g / L, 5-100 g / L, 5-75 g / L, 5-50 g / L, 5-25 g / L, 5-10 g / L, 10-100 g / L, 10-75 g / L, 10-50 g / L, 10-25 g / L, 25-100 g / L, 25-75 g / L, or 25-50 g / L. In some embodiments, this platform embodiment can generate 1-750 mg, 1-500 mg, 1-250 mg, 1-100 mg, 1-75 mg, 1-50 mg, 1-25 mg, 1-20 mg, 1-10 mg, 1-5 mg, 1-750 pg, 1-500 pg, 1-250 pg, 1-100 pg, 1-75 pg, 1-50 pg, 1-25 pg, 1-20 pg, 1-10 pg, or 1-5 pg of protein within 24-72 hours. In some embodiments, this platform can produce protein continuously for several days with continuously, or periodically, added reagents. In some embodiments, this platform can produce protein continuously for 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 days. Various parameters understood by persons of skill in the ail can be varied to achieve the yield, scale, and speed of protein preparation desired.

[0116] In some embodiments the housing comprises a polymer membrane tube, cap, and base. In some embodiments, the tube cap further comprises a hole for reagent addition. In some embodiments, polymer tube cap and base are further housed in a container. In some embodiments, the protein is translated through the polymer membrane tube into the product portion of the embodiment. In some embodiments the ribosomes in the reaction would translate the protein through the polymer membrane of the tube into the external containment vessel. In some embodiments the ribosomes are bound to the outside of the tube membrane. In these embodiments the reaction solution would translate the protein through the polymer membrane of the tube into the internal compartment of the tube. These embodiments would allow for a widerange of control over the chemical environment of the platform and allow for high purity of the proteins being translated. In some embodiments the product chamber further comprises a reaction chamber. In some embodiments reaction chamber comprises enzymes, one or more substrate, and protein synthesis reagents. In some embodiments enzymes assist in protein folding and / or post-translational modification. In some embodiments reagents comprise macromolecular crowding reagents such as liposomes.

[0117] In some embodiments protein folding enzymes include chaperone proteins such as chaperonins. In some embodiments enzymes that assist in protein folding and / or post- translational modification are selected from those described herein. In some embodiments, reaction chamber further comprises metal ions. In some embodiments metal ions are selected from Ca, Co, Cr, Cu, Fe, K, Mg, Mo, Mn, Na, Ni, V, Zn, functional equivalents, and combinations thereof.

[0118] Embodiments where the platform has a housing which comprises a polymer membrane tube, cap, and base can generate up to 100 mg of protein within 24-72 hours, depending on reaction volume and membrane surface area. In some embodiments, the target yield of protein is 1-100 g / L, 1-75 g / L, 1-50 g / L, 1-25 g / L, 1-10 g / L, 1-5 g / L, 5-100 g / L, 5-75 g / L, 5-50 g / L, 5-25 g / L, 5-10 g / L, 10-100 g / L, 10-75 g / L, 10-50 g / L, 10-25 g / L, 25-100 g / L, 25-75 g / L, or 25-50 g / L. In some embodiments, this platform embodiment can generate 1-750 mg, 1-500 mg, 1-250 mg, 1-100 mg, 1-75 mg, 1-50 mg, 1-25 mg, 1-20 mg, 1-10 mg, 1-5 mg, 1-750 pg, 1-500 pg, 1-250 pg, 1-100 pg, 1-75 pg, 1-50 pg, 1-25 pg, 1-20 pg, 1-10 pg, or 1-5 pg of protein within 24-72 hours. In some embodiments, this platform can produce protein continuously for several days with added reagents. In some embodiments, this platform can produce protein continuously for 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 days.

[0119] In some embodiments the platform housing is a vessel separated into two partitions by the polymer membrane with ribosomes attached. In these embodiments, the reaction volume and surface area can be varied by altering the dimensions of the vessel. In some embodiments the membrane is held using a support structure inserted into the vessel. In some embodiments the vessel and components are manufactured from a polymer. In some embodiments, the polymer is comprised of monomers or polymers selected from the list described herein. In some embodiments the vessel can be manufactured using various polymer manufacturing techniques including injection molding, casting, vacuum / heat forming, additive manufacturing. In someembodiments housing is sold as a kit containing the vessel, membrane and ribosomes and optionally reaction solution. In some embodiments the product chamber further comprises a reaction chamber. In some embodiments the reaction chamber comprises enzymes, substrates, and reagents.

[0120] In some embodiments, the target yield of protein for this platform embodiment is 1 g / L of reaction volume. This platform embodiment can, in various embodiments, generate up to 100 mg of protein within 24-72 hours, depending on reaction volume and membrane surface area. In some embodiments, the target yield of protein is 1-100 g / L, 1-75 g / L, 1-50 g / L, 1-25 g / L, 1-10 g / L, 1-5 g / L, 5-100 g / L, 5-75 g / L, 5-50 g / L, 5-25 g / L, 5-10 g / L, 10-100 g / L, 10-75 g / L, 10-50 g / L, 10-25 g / L, 25-100 g / L, 25-75 g / L, or 25-50 g / L. In some embodiments, this platform embodiment may generate 1-100 g, 1-75 g, 1-50 g, 1-25 g, 1-10 g, 1-5 g, 5-100 g, 5-75 g, 5-50 g, 5-25 g, 5-10 g, 10-100 g, 10-75 g, 10-50 g, 10-25 g, 25-100 g, 25-75 g, 25-50 g, 1-750 mg, 1-500 mg, 1 -250 mg, 1 -100 mg, 1 -75 mg, 1-50 mg, 1-25 mg, l-20mg, l- 10mg, l-5mg of protein within 24-72 hours. In some embodiments, this platform may produce protein continuously for several days with added reagents. In some embodiments, this platform may produce protein continuously for 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 days.

[0121] In some embodiments the platform housing is a flow reactor containing polymer membrane tubes with ribosomes attached. In some embodiments, the polymer membrane tube polymer is comprised of monomers and polymers selected from those listed herein.

[0122] In some embodiments the flow reactor can be manufactured using various polymer manufacturing techniques including injection molding, casting, vacuum / heat forming, additive manufacturing, functional equivalents, and combinations thereof. This flow reactor can be harnessed and used in industrial applications including manufacturing of therapeutics at scale, and for materials science. In some embodiments, the target yield of protein for this platform embodiment is 1 g / L of reaction volume. In some embodiments of the flow reactor, the target yield of protein is 1-100 g / L, 1-75 g / L, 1-50 g / L, 1-25 g / L, 1-10 g / L, 1-5 g / L, 5-100 g / L, 5-75 g / L, 5-50 g / L, 5-25 g / L, 5-10 g / L, 10-100 g / L, 10-75 g / L, 10-50 g / L, 10-25 g / L, 25-100 g / L, 25- 75 g / L, or 25-50 g / L. This platform embodiment can generate several kilograms of protein within 24-72 hours, depending on reaction volume and membrane surface area. In some embodiments of the flow reactor, the flow reactor can generate 1-1000 kg, 1-750 kg, 1-500 kg, 1-250 kg, 1-100 kg, 1-75 kg, 1-50 kg, 1-25 kg, 1-20 kg, 1-10 kg, 1-5 kg, 100-1000 g, 100-750 g, 100-500 g, 100-250 g, 1-100 g, 1-75 g, 1-50 g, 1-25 g, 1-10 g, 1-5 g, 5-100 g, 5-75 g, 5-50 g, 5-25 g, 5-10 g, 10- 100 g, 10-75 g, 10-50 g, 10-25 g, 25-100 g, 25-75 g, 25-50 g, of protein within 24-72 hours. In some embodiments, this platform can continuously produce protein for several days, weeks, months with continuously added reagents. In some embodiments, this platform can produce protein for 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 days, 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3- 10, 4-6, or 4-10 weeks, 1-3, 1-5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 months with continuously added reagents.

[0123] In some embodiments the platform is able to be sterilized using common industrial sterilization techniques. The specific embodiments can be selected such that to ensure reaction requirements such as turbulent flow and tangential flow are met. In some embodiments the product chamber of this platform further comprises a reaction chamber as described herein.

[0124] In some embodiments enzymes assist in protein folding, post-translational modification, macromolecular crowding reagents, liposomes. In some embodiments protein folding enzymes include chaperone proteins, chaperonins. In some embodiments enzymes are selected from Chaperonin 10, Chaperonin 60, DnaK, FIMC, GroEL, GroES, GroESL, hchA, HSCP 70, HSP32, HSP70, HSPD1, PSMG2, PSMG4, SKP, functional equivalents and combinations thereof. In some embodiments post-translational modification enzymes are selected from enzymes that perform the processes of acetylation, disulfide bond formation, glutathionylation, glycation, glycosylation, hydroxylation, lipidation, methylation, phosphorylation, proteolysis, S- nitrosylation, succinylation, SUMOylation, trisulfide bond formation, ubiquitination, functional equivalents and combinations thereof. In some embodiments macromolecular crowding reagents are selected from Dextran 70, Ficoll 70, PEG, colloid-polymer mixtures, functional equivalents and combinations thereof. In some embodiments, liposomes are prepared from cationic lipids, fatty acids, glycerolipids, ionizable lipids, PEGylated lipids, phospholipids, sterols, functional equivalents, and combinations thereof. In some embodiments, reaction chamber further comprises metal ions. In some embodiments metal ions are selected from Ca, Co, Cr, Cu, Fe, K, Mg, Mo, Mn, Na, Ni, V, Zn, functional equivalents, and combinations thereof.

[0125] In some embodiments the platform is prepared by manufacturing housing. The housing is then combined with the polymer membrane. The polymer membrane is treated with a solution of ribosomes that attach to the polymer membrane at the pores. The embodiment is then chemically stabilized using a buffer and salt solution and able to be delivered to end user. The embodimentis then combined with reaction solution. The reaction solution is then activated using mRNA encoding for the protein of interest. The reaction is allowed to proceed as long as needed / desired. The protein of interest is removed from product compartment of the platform and further processed if needed. Additional support equipment including agitators, pumps, heating and cooling tools can be sold or used alongside the platform to regulate the chemical environment.

[0126] FIG. 1 is a diagram showing the interaction and components of a ribosome with the polymer membrane, described in various embodiments, containing affinity molecules. In FIG. 1 100 is the larger ribosomal subunit, 60S in eukaryotic organisms and 50S in prokaryotic organisms. 200 is the smaller ribosome subunit 40S in eukaryotic organisms and 30S in prokaryotic. 300 is the polymer membrane, specifications of which are highly variable including but not limited to pore characteristics, material, thickness, number of layers and external support structures as described herein. 400 shows a membrane pore, characteristics of which are highly variable including width and dispersion within the membrane as described herein. 500 illustrates an affinity binding molecule attached to membrane, with high variability in ligand chemistry and other methods of binding proteins as described herein. 600 is a modified ribosomal protein designed to work with the selected affinity molecule and recombined with ribosome to form new engineered ribosome as described herein. 700 is an mRNA strand coding for the protein of interest being expressed. All enzymes needed for mRNA transcription and processing can be included in reagent mixture. 800 is a tRNA carrying amino acids and released upon transfer of amino acid to polypeptide chain. 900 is a protein of interest being expressed. 950 is polypeptide strand of protein of interest being expressed. In FIG. 1, the larger ribosome subunit

[0100] , 60S in eukaryotic organisms and 30S in prokaryotic organisms, is attached to the polymer membrane. The smaller ribosomal subunit

[0200] , 40S in eukaryotic organisms and 30S in prokaryotic organisms, associates with the larger ribosomal subunit

[0100] during the translation process. During translation, the tRNA molecules

[0800] are used up. By adding aminoacyl tRNA synthetases as described herein, the tRNA is recycled and can increase platform efficiency and yield. Within the platform, the polymer membrane

[0300] as described herein, contains an affinity molecule

[0500] as described herein, near’ the membrane pore

[0400] as described herein. This diagram shows how the ribosome [100, 200] attaches to the polymer membrane

[0300] at the affinity molecules

[0500] . The alignment of the ribosome tunnel opening over the polymer membrane pore

[0400] allows for the polypeptide strand

[0950] translated by the ribosome[100,200] to pass through the membrane pore

[0400] . By selecting which ribosomal proteins

[0600] to modify, and where affinity molecules

[0500] are on the polymer membrane

[0300] , the ribosome alignment can be tailored as needed. This will allow for new ribosomes to be incorporated within the platform. Because the polymer membrane keeps reagents separate from the protein

[0900] being translated, the platform can produce proteins with high purity.

[0127] FIG. 2 is a diagram showing the interaction and components of a ribosome with the polymer membrane tube, provided in various embodiments herein, comprising affinity molecules. 100. Larger Ribosome subunit 60S in eukaryotic organisms and 50S in prokaryotic. 200. Smaller ribosome subunit 40S in eukaryotic organisms and 30S in prokaryotic. 300. Polymer membrane tube, specifications of which are highly variable including but not limited to pore characteristics, material, thickness, number of layers and external support structures as described herein. 400. Membrane pore, characteristics of which are highly variable including width and dispersion within the membrane. 500. Affinity binding molecule attached to membrane, with high variability in ligand chemistry and other methods of binding proteins as described herein. 600. Modified ribosomal protein designed to work with the selected affinity molecule and recombined with ribosome to form new engineered ribosome. 700. mRNA strand coding for the protein of interest being expressed. All enzymes needed for mRNA transcription and processing are included in reagent mixture. 800. tRNA carrying amino acids and released upon transfer of amino acid to polypeptide chain. 900. Protein of interest being expressed. 950. Polypeptide strand of protein of interest being expressed. In FIG. 2, the larger ribosome subunit

[0100] , 60S in eukaryotic organisms and 30S in prokaryotic, is attached to the polymer membrane tube. The smaller ribosomal subunit

[0200] , 40S in eukaryotic organisms and 30S in prokaryotic, associates with the larger ribosomal subunit

[0100] during the translation process. During translation, the tRNA molecules

[0800] are used up. By adding aminoacyl tRNA synthetases as described herein, the tRNA is recycled and can increase platform efficiency and yield. Within the platform, the polymer membrane tube

[0300] as described herein, contains an affinity molecule

[0500] as described herein, near the polymer membrane pore

[0400] as described herein. This diagram shows how the ribosome [100, 200] attaches to the polymer membrane tube

[0300] at the affinity molecules

[0500] . The alignment of the ribosome tunnel opening over the polymer membrane pore

[0400] allows for the polypeptide strand

[0950] translated by the ribosome[100,200] to pass through the membrane pore

[0400] . By selecting which ribosomal proteins

[0600] to modify, and where affinity molecules

[0500] are on the polymer membrane

[0300] , the ribosome alignment can be tailored as needed. This will allow for new ribosomes to be incorporated within the platform and new affinity molecules as described herein. Because the polymer membrane keeps reagents separate from the protein

[0900] being translated, the platform can produce proteins with high purity. Because the polymer membrane is shaped into a tube, the reaction solution can be pumped through the tube in some embodiments such as the flow reactor described herein. This continual flow of reagents can produce high yield embodiments as described herein, applicable for industrial uses. Energy molecules are also shown in this diagram and are described herein. Depending on the specifics of the reaction solution, energy molecules can vary as described herein.

[0128] FIG. 3 is a diagram showing the interaction and components of a ribosome with the polymer membrane containing affinity molecules. Chemical diagram of the general mechanism of an affinity molecule binding to appropriate amino acid tag on a ribosomal protein. In this example, a Ni affinity molecule is pictured. Other chemical ligands or affinity binding methods can be used but general principle remains the same as described herein. Within the diagram, R indicates a suitable polymer that comprises the membrane as described herein. The dimensions of the polymer membrane pore are not specified as this is a variable described herein. The affinity binding molecule can be any affinity molecule described herein. The affinity binding amino acid sequence can be selected from the descriptions provided herein or sequence listing provided herein.

[0129] FIG. 4 is a diagram showing the interaction and components of a ribosome with the polymer membrane containing affinity molecules. 100. Larger Ribosome subunit 60S in eukaryotic organisms and 50S in prokaryotic. 200. Smaller ribosome subunit 40S in eukaryotic organisms and 30S in prokaryotic. 300. Polymer membrane, specifications of which are highly variable including but not limited to pore characteristics, material, thickness, number of layers and external support structures. 400. Membrane pore, characteristics of which are highly variable including width and dispersion within the membrane. 550. Antibody used to bind to ribosomal protein and attached to the membrane pore in a method similar to ELISA tests although other methods to achieve antibody attachment to a polymer can be used. 650. Modified ribosomal protein designed to work with the selected antibody molecule and recombined with ribosome to form new engineered ribosome. 700. mRNA strand coding for the protein of interest beingexpressed. All enzymes needed for mRNA transcription and processing are included in reagent mixture. 800. tRNA carrying amino acids and released upon transfer of amino acid to polypeptide chain. 900. Protein of interest being expressed. 950. Polypeptide strand of protein of interest being expressed. In FIG. 3, the larger ribosome subunit

[0100] , 60S in eukaryotic organisms and 30S in prokaryotic, is attached to the polymer membrane. The smaller ribosomal subunit

[0200] , 40S in eukaryotic organisms and 30S in prokaryotic, associates with the larger ribosomal subunit

[0100] during the translation process. During translation, the tRNA molecules

[0800] are used up. By adding aminoacyl tRNA synthetases as described herein, the tRNA is recycled and can increase platform efficiency and yield. Within the platform, the polymer membrane

[0300] as described herein, contains an affinity molecule

[0550] in this diagram an antibody, as described herein, near the membrane pore

[0400] as described herein. This diagram shows how the ribosome [100, 200] attaches to the polymer membrane

[0300] at the affinity molecules

[0550] in this diagram antibodies. The alignment of the ribosome tunnel opening over the polymer membrane pore

[0400] allows for the polypeptide strand

[0950] translated by the ribosome [100,200] to pass through the membrane pore

[0400] . By selecting which ribosomal proteins

[0650] to modify with an antibody epitope, and where affinity molecules

[0550] in this diagram antibodies, are on the polymer membrane

[0300] , the ribosome alignment can be tailored as needed. This will allow for new ribosomes to be incorporated within the platform. Because the polymer membrane keeps reagents separate from the protein

[0900] being translated, the platform can produce proteins with high purity.

[0130] FIG. 5 is a diagram showing the interaction and components of a ribosome with the polymer membrane embedded membrane protein. 100. Larger Ribosome subunit 60S in eukaryotic organisms and 50S in prokaryotic. 200. Smaller ribosome subunit 40S in eukaryotic organisms and 30S in prokaryotic. 300. Polymer membrane, specifications of which are highly variable including but not limited to pore characteristics, material, thickness, number of layers and external support structures. 450. Pore of membrane protein (integrated into the chemical structure of the polymer membrane) which allows for the polypeptide strand to lass through to product portion of system. 575. Membrane protein integrated into the chemical structure of the polymer membrane. This method closely mimics what is seen in the endoplasmic reticulum. 675. Region of the ribosomal subunit which attaches to the membrane protein to initiate translation at the endoplasmic reticulum. A hinge protein can be added if necessary to the system to mimic thenatural process of protein translation. 700. mRNA strand coding for the protein of interest being expressed. All enzymes needed for mRNA transcription and processing are included in reagent mixture. 800. tRNA carrying amino acids and released upon transfer of amino acid to polypeptide chain. 900. Protein of interest being expressed. 950. Polypeptide strand of protein of interest being expressed. In FIG. 5, the larger ribosome subunit

[0100] , 60S in eukaryotic organisms and 30S in prokaryotic, is attached to the polymer membrane via a membrane protein

[0575] as described herein. The smaller ribosomal subunit

[0200] , 40S in eukaryotic organisms and 30S in prokaryotic, associates with the larger ribosomal subunit

[0100] during the translation process. During translation, the tRNA molecules

[0800] are used up. By adding aminoacyl tRNA synthetases as described herein, the tRNA is recycled and can increase platform efficiency and yield. Within the platform, the polymer membrane

[0300] as described herein, contains an embedded membrane protein

[0575] as described herein. This diagram shows how the ribosome [100, 200] attaches to the polymer membrane

[0300] at the embedded membrane protein. The alignment of the ribosome tunnel opening over the embedded membrane protein

[0575] channel

[0450] allows for the polypeptide strand

[0950] translated by the ribosome [100,200] to pass through the membrane protein channel

[0450] . By selecting which ribosomal proteins

[0650] to modify or which ribosomes are compatible with membrane proteins, the ribosome alignment can be tailored as needed to ensure ribosome translated polypeptide through the membrane protein channel

[0450] , This will allow for new ribosomes and channel proteins, as described herein, to be incorporated within the platform. Because the polymer membrane keeps reagents separate from the protein

[0900] being translated, the platform can produce proteins with high purity. Not pictured within this diagram is a hinge protein and other molecules associated with binding the ribosome to a membrane protein embedded within the polymer membrane as described herein.

[0131] In FIG. 6 pictured are the components of a CFPS reaction kit for small scale protein expression. Dotted lines indicate assembly. 100. Housing container or vessel. Container houses the platform embodiment as described herein. 200. Cap. Cap is attached to the dialysis bag polymer membrane. Cap is hollow to allow reagents to be added. Cap can be closed off to prevent ingress of contaminants. 300. Clamp. Dialysis bag clamp or other suitable clamp is used to close off the bottom of the dialysis bag. Additional clamp can be used to close off dialysis mag instead of cap. 400. Cap holder. Cap holder contains dialysis bag with attached cap and keeps platform suspended in container or vessel solution. 500. Dialysis bag. Dialysis bag made fromthe polymer membrane with affinity molecules as described herein. The polymer membrane tube

[0500] can be clamped

[0300] at both ends and left within the container solution. This would then render the cap

[0200] and the cap holder

[0400] not necessary for use. To assemble the embodiment, the cap

[0200] is attached to the polymer membrane tube

[0500] . This assembly is then slid through the cap holder

[0400] opening. Once combined, the bottom of the tube is clamped

[0300] . A solution containing ribosomes is added to the dialysis bag and spread over the polymer membrane surface to attach ribosomes to the affinity molecules. Reaction solution is added to the dialysis bag and translation of proteins through the polymer membrane can begin. The dialysis bag embodiment is left suspended in the container

[0100] solution by the cap holder. The container solution would then comprise the product portion of the embodiment as described herein. In embodiments where a cap

[0200] and cap holder

[0400] are not used, the dialysis bag is clamped on bottom of the polymer membrane tube. The solution containing ribosomes as described herein is added to the dialysis bag and spread over the surface of the polymer membrane to attach the ribosomes to the affinity molecules as described herein. The reaction solution as described herein is added to the dialysis bag. The mRNA for the protein of interest is added to the dialysis bag. The dialysis bag is then clamped from the top using a suitable clamp

[0300] , The dialysis bag is placed in the solution inside the container

[0100] and reaction is allowed to proceed as necessary. The solution of the container

[0100] comprises the product portion of the embodiment as described herein. In a different embodiment, the polymer membrane tube is clamped from the bottom. The polymer membrane tube is slid through the cap holder

[0400] hole. The cap

[0200] is attached to the polymer membrane tube at the top. A buffer solution as described herein is added to the dialysis bag. To attach ribosomes to outside of the polymer membrane, the dialysis bag is dipped into container

[0100] solution so that ribosomes can attach to affinity molecules on the outside of the dialysis bag. Reaction solution is added to the container

[0100] along with the mRNA for the protein of interest. The reaction is allowed to proceed as needed. In this embodiment, the dialysis bag comprises the product portion of the platform as described herein. If cap

[0200] and cap holder

[0400] are not used the top of the polymer tube can be clamped using a suitable clamp

[0300] . An agitator can be added to container

[0100] to keep solution mixing.

[0132] In FIG. 7 pictured are the components of an assembled CFPS reaction kit for small scale protein expression. 100. Housing container or vessel. Container houses the platform embodimentas described herein. 200. Cap. a Cap is attached to the dialysis bag polymer membrane. The Cap is hollow to allow reagents to be added. The Cap can be closed off to prevent ingress of contaminants. 300. Clamp. Dialysis bag clamp or other suitable clamp is used to close off the bottom of the dialysis bag. Additional clamp can be used to close off dialysis bag instead of cap. 400. Cap holder. The Cap holder contains dialysis bag with attached cap and keeps platform suspended in container or vessel solution. 500. Dialysis bag. Dialysis bag made from the polymer membrane with affinity molecules as described herein. For detailed description of assembly and use of this embodiment refer to detailed description of FIG. 7.

[0133] In FIG. 8 pictured are the components of a CFPS reaction kit for small scale protein expression. Dotted lines indicate assembly. 100. Housing container or vessel. Container houses the platform embodiment as described herein. 200. Cap. Cap is attached to the polymer membrane tube. Cap can be hollow to allow reagents to be added. Hollow cap can be closed off to prevent ingress of contaminants. 300. Base. Base is attached to bottom of polymer membrane tube to seal off reaction solution. 400. Cap holder. The Cap holder contains polymer membrane tube with attached cap and keeps platform suspended in container or vessel solution. 500. Polymer membrane tube. Polymer membrane tube made from the polymer membrane with affinity molecules as described herein. To assemble the embodiment, the cap

[0200] and base

[0300] are attached to the polymer membrane tube

[0500] . This assembly is then slid through the cap holder

[0400] opening. Once combined, the assembled tube can be left suspended by the cap holder

[0400] . A solution containing ribosomes is added to the polymer membrane tube and spread over the polymer membrane surface to attach ribosomes to the affinity molecules.Reaction solution and mRNA for the protein of interest are added to the polymer membrane tube and translation of proteins through the polymer membrane can begin. The polymer membrane tube embodiment is left suspended in the container

[0100] solution by the cap holder. The container solution would then comprise the product portion of the embodiment as described herein. In another embodiment, the cap

[0200] and base

[0300] are attached to the polymer membrane tube

[0500] . A suitable buffer solution as described herein is added to the assembled polymer membrane tube. To attach ribosomes to outside of the polymer membrane, the dialysis bag is dipped into container

[0100] solution so that ribosomes can attach to affinity molecules on the outside of the dialysis bag. Reaction solution is added to the container

[0100] along with the mRNA for the protein of interest. The reaction is allowed to proceed as needed. In thisembodiment, the polymer membrane tube comprises the product portion of the platform as described herein. Agitators can be added to polymer membrane tube and reaction solution to keep solutions mixing.

[0134] In FIG. 9 Pictured are the components of an assembled CFPS reaction kit for small scale protein expression. 100. Housing container or vessel. Container houses the platform embodiment as described herein. 200. Cap. The Cap is attached to the polymer membrane tube. The Cap can be hollow to allow reagents to be added. Hollow caps can be closed off to prevent ingress of contaminants. 300. Base. Base is attached to bottom of polymer membrane tube to seal off reaction solution. 400. Cap holder. Cap holder contains polymer membrane tube with attached cap and keeps platform suspended in container or vessel solution. 500. Polymer membrane tube. Polymer membrane tube made from the polymer membrane with affinity molecules as described herein. For detailed description of assembly and use of this embodiment refer to detailed description of FIG. 8.

[0135] In FIG. 10 Pictured are components of a CFPS reaction kit for moderate scale protein expression. A membrane is not shown in this diagram. Refer to Figure 12 and Figure 13 for membrane integration. 100. Lab kit housing. Lab kit housing can have variable dimensions and material choice depending on application requirements. Although grooves pictured are continuous, they can be noncontinuous or be segmented if application requires it. Dimensions of grooves are variable as well. The shape of the housing is also able to be changed / modified per application or to adjust reaction volume / surface area ratio. 200. Perforated Frame. Perforated frame can have variable perforations that help hold the frame assembly together. Membrane not pictured in this diagram. Refer to FIG. 12 and FIG. 13 for diagrams with membrane. 250. Perforation. A perforation is made in the frame to allow for assembly of the frame and fitment. Perforations can be variable in shape, distribution and dimensions. 300. Protruded frame. Protruded frame can have variable protrusions that help hold the frame assembly together by corresponding to the perforations on the other frame component. Membrane not pictured in this diagram. Refer to FIG. 12 and FIG. 13 for diagrams with membrane. 350. Protrusion. A protrusion is made in the frame to allow for mating with corresponding perforations in the other frame component. Mating of these surfaces will allow for tight assembly of the frame and assist with alignment. Protrusions and perforations are optional if using adhesives to assemble the membrane frame. 400. Frame assembly groove. The frame assembly groove can be continuousor non-continuous depending on application requirements. This groove is optional if adhesives are used as primary method of assembly. Dimensions of the groove are highly variable. To assemble the kit, a polymer membrane is perforated in locations corresponding to the holes

[0250] and protrusions

[0350] on the frame components [200, 300]. The membrane is placed between frame components [200, 300] and pressed together. Adhesive can be used to keep components from separating. A solution containing the ribosomes described herein is placed over the membrane and spread to attach the ribosomes to the affinity molecules on the polymer membrane as described herein. An alternative is to dip the assembled frame [200, 300] in a solution containing ribosomes to attach the ribosomes to the affinity molecules on the polymer membrane as described herein. Frame [200, 300] is then slid into the grooves

[0400] . Adhesive can be used to keep the frame from moving and to seal the assembly. Reaction solution is added to the compartment with the attached ribosomes. This compartment comprises the reaction portion of the platform as described herein. A buffer, as described herein, is added to the other compartment. This compartment comprises the product portion of the platform as described herein. In the reaction compartment, mRNA for the protein of interest is added to the reaction solution and translation of protein through the polymer membrane pores can proceed as described herein. Agitators can be added to both compartments of the kit to keep solutions mixing.

[0136] FIG. 11 shows a schematic of the assembled components of a CFPS reaction kit for moderate scale protein expression. Membrane is not pictured in this diagram. Refer to FIG. 12 and FIG. 13 for membrane integration. 100. Lab kit housing. Lab kit housing can have variable dimensions and material choice depending on application requirements. Although grooves pictured are continuous, they can be noncontinuous or be segmented if application requires it. Dimensions of grooves are variable as well. The shape of the housing is also able to be changed / modified per application or to adjust reaction volume / surface area ratio. 200. Perforated Frame. Perforated frame can have variable perforations that help hold the frame assembly together. Membrane not pictured in this diagram. Refer to Figures H and I for diagrams with membrane. 250. Perforation. A perforation is made in the frame to allow for assembly of the frame and fitment. Perforations can be variable in shape, distribution and dimensions. 300. Protruded frame. Protruded frame can have variable protrusions that help hold the frame assembly together by corresponding to the perforations on the other frame component.Membrane not pictured in this diagram. Refer to Figures 12 and 13 for diagrams with membrane. 350. Protrusion. A protrusion is made in the frame to allow for mating with corresponding perforations in the other frame component. Mating of these surfaces will allow for tight assembly of the frame and assist with alignment. Protrusions and perforations are optional if using adhesives to assemble the membrane frame. 400. Frame assembly groove. The frame assembly groove can be continuous or non-continuous depending on application requirements. This groove is optional if adhesives are used as primary method of assembly. Dimensions of the groove are highly variable. For detailed description of assembly and use of this embodiment refer to detailed description of FIG.10.

[0137] In FIG. 12 pictured are components of a CFPS reaction kit for moderate scale protein expression. Membrane is pictured in this diagram. 100. Lab kit housing. Lab kit housing can have variable dimensions and material choice depending on application requirements. Although grooves pictured are continuous, they can be noncontinuous or be segmented if application requires it. Dimensions of grooves are variable as well. The shape of the housing is also able to be changed / modified per application or to adjust reaction volume / surface area ratio. 200. Perforated Frame. Perforated frame can have variable perforations that help hold the frame assembly together. 250. Perforation. A perforation is made in the frame to allow for assembly of the frame and fitment. Perforations can be variable in shape, distribution and dimensions. 300. Protruded frame. Protruded frame can have variable protrusions that help hold the frame assembly together by corresponding to the perforations on the other frame component.Membrane not pictured in this diagram. 350. Protrusion. A protrusion is made in the frame to allow for mating with corresponding perforations in the other frame component. Mating of these surfaces will allow for tight assembly of the frame and assist with alignment. Protrusions and perforations are optional if using adhesives to assemble the membrane frame. 400 Frame assembly groove. The frame assembly groove can be continuous or non-continuous depending on application requirements. This groove is optional if adhesives are used as primary method of assembly. Dimensions of the groove are highly variable. 500. Proprietary membrane. The proprietary membrane is designed to attach modified ribosomes to one side. Prior to use / addition of CFPS extract, this membrane is treated with a solution of modified ribosomes. Membrane characteristics are highly variable depending on application requirements. 550. Membrane perforations. Membrane perforations are designed to fit around frame protrusion analogous to theperforated frame. This is to help with alignment and maintain strength of the membrane. In cases where adhesives are the primary method of assembly, the perforations are optional. Perforation dimensions and distribution is to be the same to the perforated frame component. To assemble the kit, a polymer membrane

[0500] is perforated

[0550] in locations corresponding to the holes

[0250] and protrusions

[0350] on the frame components [200, 300], The membrane

[0500] is placed between frame components [200, 300] and pressed together. Adhesive can be used to keep components from separating. A solution containing the ribosomes described herein is placed over the membrane

[0500] and spread to attach the ribosomes to the affinity molecules on the polymer membrane as described herein. An alternative is to dip the assembled frame [200, 300] in a solution containing ribosomes to attach the ribosomes to the affinity molecules on the polymer membrane

[0500] as described herein. Frame [200, 300] is then slid into the grooves

[0400] . Adhesive can be used to keep the frame from moving and to seal the assembly. Reaction solution is added to the compartment with the attached ribosomes. This compartment comprises the reaction portion of the platform as described herein. A buffer, as described herein, is added to the other compartment. This compartment comprises the product portion of the platform as described herein. In the reaction compartment, mRNA for the protein of interest is added to the reaction solution and translation of protein through the polymer membrane pores can proceed as described herein. Agitators can be added to both compartments of the kit to keep solutions mixing.

[0138] In FIG. 13 pictured are components of a CFPS reaction kit for moderate scale protein expression. Membrane is pictured in this diagram. 100. Lab kit housing. Lab kit housing can have variable dimensions and material choice depending on application requirements. Although grooves pictured are continuous, they can be noncontinuous or be segmented if application requires it. Dimensions of grooves are variable as well. The shape of the housing is also able to be changed / modified per application or to adjust reaction volume / surface area ratio. 200. Perforated Frame. Perforated frame can have variable perforations that help hold the frame assembly together. 250. Perforation. A perforation is made in the frame to allow for assembly of the frame and fitment. Perforations can be variable in shape, distribution and dimensions. 300. Protruded frame. Protruded frame can have variable protrusions that help hold the frame assembly together by corresponding to the perforations on the other frame component. 350. Protrusion. A protrusion is made in the frame to allow for mating with correspondingperforations in the other frame component. Mating of these surfaces will allow for tight assembly of the frame and assist with alignment. Protrusions and perforations are optional if using adhesives to assemble the membrane frame. 400. Frame assembly groove. The frame assembly groove can be continuous or non-continuous depending on application requirements. This groove is optional if adhesives are used as primary method of assembly. Dimensions of the groove are highly variable. 500. Proprietary membrane. The proprietary membrane is designed to attach modified ribosomes to one side. Prior to use / addition of CFPS extract, this membrane is treated with a solution of modified ribosomes. Membrane characteristics are highly variable depending on application requirements. 550. Membrane perforations. Membrane perforations are designed to fit around frame protrusion analogous to the perforated frame. This is to help with alignment and maintain strength of the membrane. In cases where adhesives are the primary method of assembly, the perforations are optional. Perforation dimensions and distribution is to be the same to the perforated frame component. For detailed description of assembly and use of this embodiment refer to detailed description of FIG.12.

[0139] In FIG. 14 pictured are the components of a flow reactor for large scale protein expression. 100. Chamber. The chamber dimensions and shape are variable per application needs. Polymer material choice is described herein. Ideally chamber dimensions and materials are selected for pressure differential resistance and sterilizability. 200. Perforated base plate. The base plate has perforations to hold the membrane tubes and to provide separation between the reaction and product portions of the embodiment as described herein. Perforation shape and dimensions are variable. Base plate dimensions, including perforation distribution and amount, and material choice are designed as needed per the end user requirements. Edges of perforations can be modified onto a cone or other shape to change flow and turbulence and can have a protrusion to better hold the polymer membrane tube. 300. Chamber cap. The chamber cap is designed with variability in materials, dimensions, edges, and shape to optimize for the end user requirements. Chamber cap includes a process connection to connect with industrial plumbing. The chamber cap is attached to the outside of the chamber and has an edge to hold baseplate. 400. Membrane tube. The membrane tube is designed to separate reaction solution from the product portion, and to bind the ribosome to the affinity molecules on the polymer membrane as described herein. In some embodiments the polymer material selected will allow for resistance to a pressure differential and sterilizability. The protein of interest is expressed through themembrane tube pores into the chamber which comprises the product portion of the platform. 500. Process connection. The process connections are there to readily integrate the flow reactor into industry standard connections and allow for use with regular process plumbing. Other connection types can be used including threaded fittings and other commonly used connections in industry. To assemble the flow reactor, the base plates

[0200] are attached to the chamber

[0100] top and bottom. Components can be attached together using adhesives, cement or heat. The polymer membrane tubes

[0400] as described herein, are slid through the perforations on the top and bottom base plates

[0200] . The polymer membrane tubes can be held in place using adhesives. Adhesives will also provide a seal separating the flow reactor into a reaction and product compartment. The chamber caps

[0300] are attached to the top and bottom of the assembled chamber. Components can be attached together using adhesives, cement or heat. To use the flow reactor, the flow reactor is connected to a vessel containing a solution of ribosomes as described herein. This solution is pumped through the chamber connections

[0500] and through the inside of the polymer membrane tubes with affinity molecules described herein. A vessel containing a buffer solution described herein is connected to the process connections

[0500] of the flow reactor chamber

[0100] , Buffer solution described herein is pumped through the chamber process connections

[0500] to create a product partition of the embodiment as described herein. To use the flow reactor, the reaction solution described herein in the vessel connected to the chamber cap

[0300] connections

[0500] along with the mRNA for the protein of interest are pumped through the flow reactor through the polymer membrane tubes

[0400] containing bound ribosomes. The ribosomes translate the protein of interest through the pores on the polymer membrane tube into the product partition of the flow reactor as described herein. The reaction proceeds as determined by end user as described herein. In another embodiment, the ribosomes are attached to the outside of the polymer membrane tube by utilizing the chamber

[0100] process connection in which case the reaction and product partition are inverted.

[0140] In FIG. 15 pictured is an assembled flow reactor for large scale protein expression. 100. Chamber. The chamber dimensions and shape are variable per application needs. Polymer material choice is described herein. Ideally chamber dimensions and materials are selected for pressure differential resistance and sterilizability. 200. Perforated base plate. The base plate has perforations to hold the membrane tubes and to provide separation between the reaction and product portions of the embodiment as described herein. Perforation shape and dimensions arevariable. Base plate dimensions, including perforation distribution and amount, and material choice are designed as needed per the end user requirements. Edges of perforations can be modified onto a cone or other shape to change flow and turbulence and can have a protrusion to better hold the polymer membrane tube. 300. Chamber cap. The chamber cap is designed with variability in materials, dimensions, edges, and shape to optimize for the end user requirements. Chamber cap includes a process connection to connect with industrial plumbing. The chamber cap is attached to the outside of the chamber and has an edge to hold baseplate. 400. Membrane tube. The membrane tube is designed to separate reaction solution from the product portion, and to bind the ribosome to the affinity molecules on the polymer membrane as described herein. In some embodiments the polymer material selected will allow for resistance to a pressure differential and sterilizability. The protein of interest is expressed through the membrane tube pores into the chamber which comprises the product portion of the platform. 500. Process connection. The process connections are there to readily integrate the flow reactor into industry standard connections and allow for use with regular process plumbing. Other connection types can be used including threaded fittings and other commonly used connections in industry. For detailed description of assembly and use of this embodiment refer to detailed description of FIG.14.

[0141] In FIG. 16 pictured is the Candida albicans 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L26B, L31B, L35A are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0142] In FIG. 17 pictured is the Danio rerio 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L31 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane atthe affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0143] In FIG. 18 pictured is the Drosophila melanogaster 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L26, L31, L35 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0144] In FIG. 19 pictured is the Escherichia coli 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22, L23, L24, L29 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0145] In FIG. 20 pictured is the Haloarcula marismortui 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22P, L23P, L24P, L29P, L31e are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0146] In FIG. 21 pictured is the Homo sapiens 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L26, L31, L35 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0147] In FIG. 22 pictured is the Kluyveromyces lactis 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L25, KLLA0A06336p, KLLA0B02937p, KLLA0B05742p, KLLA0E12453p, KLLA0F05247p are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0148] In FIG. 23 pictured is the Mas musculus 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L19, L26, L31, L35 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0149] In FIG. 24 pictured is the Nicotiana tabacum 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22, L22-2, L23a, L26, L31, L35, L38 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind theaffinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0150] In FIG. 25 pictured is the Oryctolagus cuniculus 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L24, L29, SRP19, SRP54, are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0151] In FIG. 26 pictured is the Pseudomonas aeruginosa 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L23, L24, L29, L32 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0152] In FIG. 27 pictured is the Pyrococcus furiosus 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L3P, L24P, L29P, L31e are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosomethrough the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0153] In FIG. 28 pictured is the Saccharomyces cerevisiae 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L17, L22, L23, L24, L29 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0154] In FIG. 29 pictured is the Spinacia oleracea 50S Chloroplast ribosomal subunit structure from the outside perspective. Ribosomal proteins rpL17, rpL22, rpL26, rpL31 , rpL38 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0155] In FIG. 30 pictured is the Staphylococcus aureus 50S ribosomal subunit structure from the outside perspective. Ribosomal proteins L22, L23, L24, L29 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0156] In FIG. 31 pictured is the Sus scrofa 60S ribosomal subunit structure from the outsideperspective. Ribosomal proteins eL22, uL22, L23, L24, L29, L31, L38 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.

[0157] In FIG. 32 pictured is the Thermus thermophilus 60S ribosomal subunit structure from the outside perspective. Ribosomal proteins L23, L24, L29, L32 are highlighted. These ribosomal proteins can be modified to contain ligand binding sites suitable to bind the affinity binding molecules on the polymer membrane. Refer to Table 1. for the list of ribosomal protein modifications. The ribosome tunnel opening is also visible and labeled. When attached to a polymer membrane at the affinity molecules as described herein, the ribosome tunnel opening aligns with the polymer membrane pore so that the polypeptide is translated by the ribosome through the polymer membrane pore as described herein. Combinations of affinity molecules can be selected to better align the ribosome tunnel opening over the polymer membrane pore.EXAMPLES

[0158] Cell - free systems have mostly been developed using E. coli and other model organism extracts until recently. This is significant because the scope of extract-based Cell-Free Protein Synthesis (CFPS) applications is determined by the chosen source organism and the biochemical resources present at the time of cell harvest and extract preparation. This means that metabolic enzymes, cellular machinery like the translation system, and co - factor and energy regeneration systems, unique to a chassis organism are also unique to that organism's extract. The most exploited CFPS systems are from E. coli (bacterium) (Carlson et al., 2012; Hodgman and Jewett, 2012), wheat germ (plant) (Madin et al., 2000; Takai et al., 2010), Spodoptera frugiperda (insect) (Ezure et al., 2010; Tarui et al., 2001), and rabbit reticulocytes (mammal) (Anastasina et al., 2014; Kobs, 2008; Pelham and Jackson, 1976), with others being developed (Ferrer-Miralles et al., 2009; Gan and Jewett, 2014; Hodgman and Jewett, 2013), (Kovtunet al., 2010; Mureev et al.,2009), (Brodel et al., 2014; Martin et al., 2017), (Wang et al., 2018), (Mikami et al., 2010), (Kelwick et al., 2016). However, CFPS systems derived from non-model organisms have only recently been developed, most notably from species of archeae (Endoh et al., 2008, 2007, 2006), Bacillus (Moore, Simon J.; MacDonald, James T.; Wienecke, Sarah; Ishwarbhai, Alka; Tsipa, Argyro; Aw, Rochelle; Kylilis, Nicolas; Bell, David J., McClymont, David W.; Jensen, Kirsten; Polizzi, Karen M.; Biedendieck, Rebekka; Freemont, 2018), Streptomyces (Li et al., 2018, 2017), and Vibrio (Des Soye et al., 2018 ; Failmezger et al., 2018; Wiegand et al., 2018 ).

[0159] Example 1: Preparation of Platform Synthesis of Polymer Membrane

[0160] The polymer membrane used within the platform can be prepared using the following methods:

[0161] Method 1. Spraying: A dendrimer of relevant dimensions is synthesized. Prepared dendrimers or custom dendrimers can be ordered from Bio-Synthesis, CD Bioparticles, Creative Biolabs, Dendritech, Life Chemicals, NARD Institute, Richmann Chemical. Dendrimer is reacted using chemical methods with an affinity molecule as aforementioned. Reaction is performed in a way so that the affinity molecule attached to the dendrimer can react at its base with PES monomers and solvent. Dendrimer is distributed on a prepared surface using spraying. Spraying conditions are adjusted to change distribution of dendrimers on prepared surface. Care is taken to ensure monolayer distribution of dendrimers. PES monomer, purchasable from Fisher Scientific, Richmann Chemical, Sigma Aldrich, is dissolved in an appropriate solvent purchasable from Richmann Chemical, Sigma Aldrich. Solvent is selected to ensure nonreactivity with dendrimers on prepared surface. Appropriate solvent, containing monomers, is sprayed onto prepared surface containing dendrimers. Care is taken to ensure PES layer thickness is not thicker than dimensions of dendrimers. Solvent is allowed to evaporate to polymerize PES monomers. Care is taken to ensure affinity molecules on dendrimers are able to polymerize with PES monomers. Drying of newly formed PES membrane is performed to evaporate residual solvent. Solvent is selected that is able to dissolve the aforementioned dendrimer but leave PES polymer and affinity molecules intact. Membrane is studied using SEM to ensure proper chemical characteristics, pore dimensions and distribution are achieved. PESmembrane with affinity molecules is then formed into a tube, bag of appropriate dimensions or is kept flat.

[0162] Method 2. Casting: Casting of membranes is a common method to manufacture polymer membranes. This method is also known as phase separation. A dendrimer of relevant dimensions is synthesized. Prepared dendrimers or custom dendrimers can be ordered from Bio-Synthesis, CD Bioparticles, Creative Biolabs, Dendritech, Life Chemicals, NARD Institute, Richmann Chemical. Dendrimer is reacted using chemical methods with an affinity molecule as aforementioned. Reaction is performed in a way so that the affinity molecule attached to the dendrimer can react at its base with PES monomers and solvent. Dendrimer is distributed on a solvent that is inorganic such as water. Care is taken to ensure dendrimers are spread in an even single layer. An organic solution containing PES monomer is poured onto the water layer. Monomer solution is then spread into a thin layer. Solution is kept long enough to form a polymer membrane layer with thickness of the dendrimers. Affinity molecule monomer is polymerized with the PES monomers during this phase. Membrane is then removed from solution and dried. Solvent is added to dissolve dendrimers leaving the affinity molecules near the polymer membrane pore edge.Preparation of Reaction Solution

[0163] A Cell-free protein synthesis reaction solution can be purchased from a variety of vendors. For example, a cell-free protein synthesis reaction solution can be purchased from NE Biolabs (PURExpress® A Ribosome Kit) that does not contain reconstituted ribosomes. Cell-free protein synthesis extracts are well described in literature including preparation methods. A cell culture is grown. Cells are lysed and solution is centrifuged to remove cellular debris. Solution is then purified using various columns and concentrated. RNAases are added to remove residual genetic material. TRNAs are added to solution along with buffers, amino acids and tRNA aminoacyl synthetases to recycle tRNA molecules. The Solution is then ready to be added to the platform.

[0164] Example 2:Using Platform

[0165] To attach ribosomes to the polymer membrane tube, dialysis bag or flat membrane, a solution containing modified ribosomes described herein is added to the polymer tube, dialysis bag or flat membrane partition. The polymer tube, dialysis bag, or flat membrane partition is thenplaced horizontally and rolled to distribute ribosomes along affinity molecules on polymer membrane tube pores. To attach ribosomes to polymer membrane tubes in the flow reactor, ribosome containing solution is pumped through the flow reactor.

[0166] Reaction solution described herein is added to the polymer membrane tube, dialysis bag, partitioned vessel, flow reactor. Reaction solution is purchasable commercially from New England Biolabs (PURExpress® A Ribosome Kit).

[0167] MRNA for protein of interest is added to reaction solution along with energy molecules due to their inherent instability in solution. Reaction is allowed to proceed as long as deemed necessary by end user.

[0168] Enzymes are added to the product portion of the platform that perform acetylation, disulfide bond formation, glutathionylation, glycation, glycosylation, hydroxylation, lipidation, methylation, phosphorylation, proteolysis, S-nitrosylation, succinylation, SUMOylation, trisulfide bond formation, ubiquitination and other post-translation modification processes. These enzymes are readily available for purchase from Abeam, Jena Bioscience, SigmaAldrich, ThermoFisher. Chaperonins are added to the product portion of the platform to assist in protein folding and are readily available for purchase from CreativeBiomart, Fisher Scientific, Molecular Depot, MyBioSource, SigmaAldrich, StressMarq, Syd Labs. Substrate is added to assist in protein folding including enzymes, reagents, and ions. Liposomes are added to the product portion of the platform to allow expression of membrane proteins. Lipids used to synthesize liposomes are purchasable form Advansta, Avanti Research, BOC Sciences, BroadPharm, Fisher Scientific, Hzymes, LabCart, Kyfora Bio, Polysciences, Proteintech Group, Sigma Aldrich. Such additions have been documented in literature, and one skilled in the ail would be able to tailor the platform to individual needs.

[0169] Solution from product compartment is removed and centrifugated to concentrate protein produced by the platform. Any necessary purification and concentration steps are performed to further purify the protein.

[0170] Example 3:

[0171] Dendrimers can be used to bind to biological molecules including nucleic acids as discussed herein. This is an active field of research particularly in pharmacology as dendrimers are useful in delivering therapeutics. An example of literature discussing the use of dendrimers for drug delivery purposes (Jiayi Pan, 2020). Jiayi Pan, S. A. (2020). Dendrimers for drugdelivery purposes. In J. Pan, Nanoengineered Biomaterials for Advanced Drug Delivery (pp. 201-242). Elsevier.

[0172] Methods of synthesizing dendrimers are well known to those skilled in the ail as there is significant research within the field. An example of literature discussing an approach to dendrimer synthesis. See, for example, A. Ya-Ting Huang, C.-L. K. (2023, January). Solid-phase dendrimer synthesis: a promising approach to transform dendrimer construction. Materials Today Chemistry, 27, p. 101285. This is an area where new methods to synthesize dendrimers are being continuously developed within material science and other fields of chemistry.

[0173] Polymer membrane manufacturing methods are well known to those skilled in the ail and are well described in literature. An example of literature reviewing porous polymeric membrane preparation is XueMei Tan, D. R. (2019, July 8). A Review on Porous Polymeric Membrane Preparation. Pail I: Production Techniques with Poly sulfone and Poly (Vinylidene Fluoride). Polymers, 11 (7), p. 1160. Another example of literature reviewing modified polyether-sulfone membrane is Noof A. Alenazi, M. A. (2017). Modified polyether-sulfone membrane: a mini review. Designed Monomers and Polymers, 20(1), pp. 532-546. Methods to synthesize polymer membrane are being continuously improved upon and new methods developed.

[0174] Dendrimers can be used within the polymer membrane manufacturing process. An example of literature reviewing the use of dendrimers within the preparation and modification of membranes is O. Karatas, R. K. (2022, March). A review on dendrimers in preparation and modification of membranes: progress, applications, and challenges. Materials Today Chemistry, 23, p. 100683. New methods incorporating dendrimers within manufacturing of polymer membrane are being developed as this is an ongoing field of research.

[0175] Table 1. Ribosomal Protein Modifications

[0176] This table organizes several organisms’ ribosomes that can be modified to potentially achieve binding to membrane pore. Organisms are representative of different classes and kingdoms. Persons of skill in the ail understand that additional modifications will become available for use as the research continues to develop in this field. Table 1 is organized according to organism class, species, ribosome, ribosomal proteins and ribosomal protein modifications.

[0177] Table 2. Protein Binding Methods

[0178] This table organizes some of the common methods for binding proteins. The list of protein tags includes affinity binding protein tags, antibody binding protein tags and other commonly used and available tags.

[0179] The following sequences are modifications of ribosomal proteins that can be used with various embodiments described herein.

[0180] SEQ ID NO: 1MAKISQDVSSSRSKARKAYFTASSVERRVLLSAPLSKELRQQYNVKSLPIRQNDHVLVVRGSKKHSEGKVNSVYRLKFAIQVDKLQKEKSNHASVPINIHPSKVVITKLHLDKDRKALIQRKGGKAE

[0181] SEQ ID NO: 2MAKISQDVSSSRSKARKAYFTASSVERRVLLSAPLSKELRQQYNVKSLPIRQNDEVLVVRGSKKESEGKVNSVYRLKFAIQVDKLQKEKSNEASVPINIHPSKVVITKLHLDKDRKALIQ RKGGKAE

[0182] SEQ ID NO: 3MAK1SQDVSSSRSKARKAYFTASSVERRVLLSAPLSKELRQQYNVKSLP1RQNDRVLVVRGSKKRSEGKVNSVYRLKFAIQVDKLQKEKSNRASVPINIHPSKVVITKLHLDKDRKALIQRKGGKAE

[0183] SEQ ID NO: 4MAKISQDVSSSRSKARKAYFTASSVERRVLLSAPLSKELRQQYNVKSLPIRQNDCVLVVRGSKKCSEGKVNSVYRLKFAIQVDKLQKEKSNCASVPINIHPSKVVITKLHLDKDRKALIQRKGGKAE

[0184] SEQ ID NO: 5MAHHHHVTREYTINLHKRLHGVNFKKRAPKAVKEIKKFATLHMGTTDVRLDPKLNIAIWKRGVQGVENRMRLRISRKHNHEHDAKEKLFAYVEPVIVPSTKGLQTVVVHHHH

[0185] SEQ ID NO: 6MAHQHQVTREYTINLHKRLHGVNFKKRAPKAVKEIKKFATLHMGTTDVRLDPKLNIAIWKRGVQGVENRMRLRISRKRNDEEDAKEKLFAYVEPVIVPSTKGLQTVVVHQHQ

[0186] SEQ ID NO: 7MAEEEEVTREYTINLHKRLHGVNFKKRAPKAVKEIKKFATLHMGTTDVRLDPKLNIAIWKRGVQGVENRMRLRISRKENEEEDAKEKLFAYVEPVIVPSTKGLQTVVVEEEE

[0187] SEQ ID NO: 8MARRRRVTREYTINLHKRLRGVNFKKRAPKAVKEIKKFATLHMGTTDVRLDPKLNIAIWKRGVQGVENRMRLRISRKRNDEEDAKEKLFAYVEPVIVPSTKGLQTVVVRRRR

[0188] SEQ ID NO: 9MACCCCVTREYTINLHKRLHGVNFKKRAPKAVKEIKKFATLHMGTTDVRLDPKLNIAIWKRGVQGVENRMRLRISRKCNCECDAKEKLFAYVEPVIVPSTKGLQTVVVCCCC

[0189] SEQ ID NO: 10MALQDVVTREYTINLHKRLHGVNFKKRAPKAVKEIKKFATLHMGTTDVRLDPKLNIAIWKRGVQGVENRMRLRISRKRNDEEDAKEKLFAYVEPVIVPSTKGLQTVVVEPEA

[0190] SEQ ID NO: 11MAGVKTFELRHHSKEQLESQLVELKQELATLKVQKLHHPSLPRIHTVRKNIARVLTVINLNQRENVRAFYAGKKYIPKDLRAKKTRALRRKLTKFEASQETEKARKQRIAFPQRKFAIK A

[0191] SEQ ID NO: 12MAGVKTFELRHQSKEQLESQLVELKQELATLKVQKLHQPSLPRIHTVRKNIARVLTVINLNQRENVRAFYAGKKY1PKDLRAKKTRALRRKLTKFEASQETEKARKQR1AFPQRKFA1KA

[0192] SEQ ID NO: 13MAGVKTFELREESKEQLESQLVELKQELATLKVQKLEEPSLPRIHTVRKNIARVLTVINLNQRENVRAFYAGKKYIPKDLRAKKTRALRRKLTKFEASQETEKARKQRIAFPQRKFAIKA

[0193] SEQ ID NO: 14MAGVKTFELRRRSKEQLESQLVELKQELATLKVQKLRRPSLPRIHTVRKNIARVLTVINLNQRENVRAFYAGKKYIPKDLRAKKTRALRRKLTKFEASQETEKARKQRIAFPQRKFAIK A

[0194] SEQ ID NO: 15MAGVKTFELRCCSKEQLESQLVELKQELATLKVQKLCCPSLPRIHTVRKNIARVLTVINLNQRENVRAFYAGKKYIPKDLRAKKTRALRRKLTKFEASQETEKARKQRIAFPQRKFAIK A

[0195] SEQ ID NO: 16MVRYSLDPENPHHSCKSRGSNLRVHFKNTRETAQAIKGMHIRKANKYLKDVIVKHQCVPFRRYNGGVGRCAQAKQHDWTQGRWPKKSAEFLLHMLKNAESNAELKHLHVHHLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVSQKKKVSQKKLKKQKLMARE

[0196] SEQ ID NO: 17MVRYSLDPENPHQSCKSRGSNLRVHFKNTRETAQAIKGMHIRKANKYLKDVIVKHQCVPFRRYNGGVGRCAQAKQHDWTQGRWPKKSAEFLLHMLKNAESNAELKHLQVHQLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVSQKKKVSQKKL10KKQKLMARE

[0197] SEQ ID NO: 18MVRYSLDPENPEESCKSRGSNLRVHFKNTRETAQAIKGMHIRKANKYLKDVIVKHQCVPFRRYNGGVGRCAQAKQHDWTQGRWPKKSAEFLLHMLKNAESNAELKELEVEELVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVSQKKKVSQKKLKKQKLMARE

[0198] SEQ ID NO: 19MVRYSLDPENPRRSCKSRGSNLRVHFKNTRETAQAIKGMHIRKANKYLKDVIVKHQCVPFRRYNGGVGRCAQAKQHDWTQGRWPKKSAEFLLHMLKNAESNAELKRLRVRRLVIEHIQVNKAPKMRRRTYRAHGR1NPYMSSPCH1EM1LTEKEQ1VPKPEEEVSQKKKVSQKKLKKQKLMARE

[0199] SEQ ID NO: 20MVRYSLDPENPCCSCKSRGSNLRVHFKNTRETAQAIKGMHIRKANKYLKDVIVKHQCVPFRRYNGGVGRCAQAKQHDWTQGRWPKKSAEFLLHMLKNAESNAELKCLCVCCLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVSQKKKVSQKKLKKQKLMARE

[0200] SEQ ID NO: 21MRFFSSSFLPKLSAMAPIKKQVTKGGKKKKQVLKFTLDCTHPVEHHIMDAANFEQFLQERIKVNGKAGNLGGGVVSIERHHSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANTKESYELRYFQINQDEEEEDED

[0201] SEQ ID NO: 22MRFFSSSFLPKLSAMAPIKKQVTKGGKKKKQVLKFTLDCTHPVEHQIMDAANFEQFLQERIKVNGKAGNLGGGVVSIERHQSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANTKESYELRYFQINQDEEEEDED

[0202] SEQ ID NO: 23MRFFSSSFLPKLSAMAPIKKQVTKGGKKKKQVLKFTLDCTHPVEEEIMDAANFEQFLQERIKVNGKAGNLGGGVVSIEREESKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANTKESYELRYFQINQDEEEEDED

[0203] SEQ ID NO: 24MRFFSSSFLPKLSAMAPIKKQVTKGGKKKKQVLKFTLDCTHPVERRIMDAANFEQFLQERIKVNGKAGNLGGGVVSIERRRSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANTKESYELRYFQINQDEEEEDED

[0204] SEQ ID NO: 25MRFFSSSFLPKLSAMAPIKKQVTKGGKKKKQVLKFTLDCTHPVECCIMDAANFEQFLQERIKVNGKAGNLGGGVVSIERCCSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANTKESYELRYFQINQDEEEEDED

[0205] SEQ ID NO: 26MAPKKGEKKKGRSAIHHHVTREYTINIHKRIHGISFKRRAPRALKEIRKFAVKEMGTPDVRIDTRLNKAVWAKGVRNVPYRMRVRLSRKHNHHEHSPNHLYTLVTYVPVTTYKGLQTV NVDEN

[0206] SEQ ID NO: 27MAPKKGEKKKGRSAIHQHVTREYTINIHKRIHGISFKRRAPRALKEIRKFAVKEMGTPDVRIDTRLNKAVWAKGVRNVPYRMRVRLSRKQNHQEHSPNKLYTLVTYVPVTTYKGLQTV NVDEN

[0207] SEQ ID NO: 28MAPKKGEKKKGRSAIEEEVTREYTINIHKRIHGISFKRRAPRALKEIRKFAVKEMGTPDVRIDTRLNKAVWAKGVRNVPYRMRVRLSRKENEEEESPNKLYTLVTYVPVTTYKGLQTVNVDEN

[0208] SEQ ID NO: 29MAPKKGEKKKGRSAIRRRVTREYTINIHKRIHGISFKRRAPRALKEIRKFAVKEMGTPDVRIDTRLNKAVWAKGVRNVPYRMRVRLSRKRNRRERSPNKLYTLVTYVPVTTYKGLQTVNVDEN

[0209] SEQ ID NO: 30MAPKKGEKKKGRSAICCCVTREYTINIHKRIHGISFKRRAPRALKEIRKFAVKEMGTPDVRIDTRLNKAVWAKGVRNVPYRMRVRLSRKCNCCECSPNKLYTLVTYVPVTTYKGLQTV NVDEN

[0210] SEQ ID NO: 31MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLHHHRLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKEHHHHHHTDDEPAKKKLSKKK LQRQKEKMLRSE

[0211] SEQ ID NO: 32MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLHQHRLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKEHQHQHQTDDEPAKKKLSKKKLQRQKEKMLRSE

[0212] SEQ ID NO: 33MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLDADRLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKEDDDDKATDDEPAKKKLSKKKLQRQKEKMLRSE

[0213] SEQ ID NO: 34MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLEEERLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKEEEEEEETDDEPAKKKLSKKKLQRQKEKMLRSE

[0214] SEQ ID NO: 35MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLRRRRLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKERRRRRRTDDEPAKKKLSKKKLQRQKEKMLRSE

[0215] SEQ ID NO: 36MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLDADRLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKEECCCCATDDEPAKKKLSKKKLQRQKEKMLRSE

[0216] SEQ ID NO: 37MGRYSRESDNVAKSCKARGPNLRVHFKNTHETAQAIKRMPLRRAQRYLKAVIDQKECVPFRRFNGGVGRCAQAKQWKTTQGRWPKKSAEFLLQLLRNAEANADCKGLDADRLVVHHIQVNRAQCLRRRTYRAHGRINPYMSSPCHVEVILTEKECCPGCCTDDEPAKKKLSKKKLQRQKEKMLRSE

[0217] SEQ ID NO: 38MAPTAKTNKGDTKTAAAKPAEKKAAPAAAAAKGKVEKPKAEAAKPAAAAAKNVKKASEAAKDVKAAAAAAKPAAAKPAAAKPAAASKDAGKKAPAAAAPKKDAKAAAAPAPAKAAPAKKAASTPAAAPPAKKAAPAKAAAPAAAAPAPAAAAPAVAKPAPKPKAKAAPAPSKVVKKNVLRGKGQKKKKVSLRFTIHCHHIAHHSIMDVADFEKYIKARLKVNGKVNNLGNNVTFERHHHHLIVSSDVHFSKAYLKYLTKKYLKKNSLRDWIRVVANEKDSYELRYFRIS SNDDEDDDAE

[0218] SEQ ID NO: 39MAPTAKTNKGDTKTAAAKPAEKKAAPAAAAAKGKVEKPKAEAAKPAAAAAKNVKKASEAAKDVKAAAAAAKPAAAKPAAAKPAAASKDAGKKAPAAAAPKKDAKAAAAPAPAKAAPAKKAASTPAAAPPAKKAAPAKAAAPAAAAPAPAAAAPAVAKPAPKPKAKAAPAPSKVVKKNVLRGKGQKKKKVSLRFT1HCQH1AQHS1MDVADFEKY1KARLKVNGKVNNLGNNVTFERQHQHLIVSSDVHFSKAYLKYLTKKYLKKNSLRDWIRVVANEKDSYELRYFRISSNDDEDDDAE

[0219] SEQ ID NO: 40MAPTAKTNKGDTKTAAAKPAEKKAAPAAAAAKGKVEKPKAEAAKPAAAAAKNVKKASEAAKDVKAAAAAAKPAAAKPAAAKPAAASKDAGKKAPAAAAPKKDAKAAAAPAPAKAAPAKKAASTPAAAPPAKKAAPAKAAAPAAAAPAPAAAAPAVAKPAPKPKAKAAPAPSKVVKKNVLRGKGQKKKKVSLRFTIECEEIAEESIMDVADFEKYIKARLKVNGKVNNLGNNVTFEREEEELIVSSDVHFSKAYLKYLTKKYLKKNSLRDWIRVVANEKDSYELRYFRISSNDDEDDDAE

[0220] SEQ ID NO: 41MAPTAKTNKGDTKTAAAKPAEKKAAPAAAAAKGKVEKPKAEAAKPAAAAAKNVKKASEAAKDVKAAAAAAKPAAAKPAAAKPAAASKDAGKKAPAAAAPKKDAKAAAAPAPAKAAPAKKAASTPAAAPPAKKAAPAKAAAPAAAAPAPAAAAPAVAKPAPKPKAKAAPAPSKVVKKNVLRGKGQKKKKVSLRFTIRCRRIARRSIMDVADFEKYIKARLKVNGKVNNLGNNVTFERRRRRLIVSSDVHFSKAYLKYLTKKYLKKNSLRDWIRVVANEKDSYELRYFRIS SNDDEDDDAE

[0221] SEQ ID NO: 42MAPTAKTNKGDTKTAAAKPAEKKAAPAAAAAKGKVEKPKAEAAKPAAAAAKNVKKASEAAKDVKAAAAAAKPAAAKPAAAKPAAASKDAGKKAPAAAAPKKDAKAAAAPAPAKAAPAKKAASTPAAAPPAKKAAPAKAAAPAAAAPAPAAAAPAVAKPAPKPKAKAAPAPSKVVKKNVLRGKGQKKKKVSLRFTICCCCIACCSIMDVADFEKYIKARLKVNGKVNNLGNNVTFERCCCCLIVSSDVHFSKAYLKYLTKKYLKKNSLRDWIRVVANEKDSYELRYFRISSNDDEDDDAE

[0222] SEQ ID NO: 43MKQNPFVSSSRRKNRKRHFQAPSHIRRRLMSAPLSKELRQKYNVRSMPIRRDDEVQVIRGHFKHHHHGKVVQAYRKKFVVYVEKIQRHNANHTNVYVGIHPSKVLIVKHHHDKDRKAILERRGKGRLAALGKDKGKYTEETAAQPMETA

[0223] SEQ ID NO: 44MKQNPFVSSSRRKNRKRHFQAPSHIRRRLMSAPLSKELRQKYNVRSMPIRRDDEVQVIRGHFKHQHQGKVVQAYRKKFVVYVEKIQRHNANQTNVYVGIHPSKVLIVKHQHDKDRKA1LERRGKGRLAALGKDKGKYTEETAAQPMETA

[0224] SEQ ID NO: 45MKQNPFVSSSRRKNRKRHFQAPSHIRRRLMSAPLSKELRQKYNVRSMPIRRDDEVQVIRGHFKEEEEGKVVQAYRKKFVVYVEKIQRENANETNVYVGIHPSKVLIVKEEEDKDRKAILERRGKGRLAALGKDKGKYTEETAAQPMETA

[0225] SEQ ID NO: 46MKQNPFVSSSRRKNRKRHFQAPSHIRRRLMSAPLSKELRQKYNVRSMPIRRDDEVQVIRGHFKRRRRGKVVQAYRKKFVVYVEKIQRRNANRTNVYVGIHPSKVLIVKRRRDKDRKAILERRGKGRLAALGKDKGKYTEETAAQPMETA

[0226] SEQ ID NO: 47MKQNPFVSSSRRKNRKRHFQAPSHIRRRLMSAPLSKELRQKYNVRSMPIRRDDEVQVIRGHFKCCCCGKVVQAYRKKFVVYVEKIQRCNANCTNVYVGIHPSKVLIVKCCCDKDRKAILERRGKGRLAALGKDKGKYTEETAAQPMETA

[0227] SEQ ID NO: 48MTKTKGEKINKSAHHHHHTRECTIHLAKRVHNIGFKKRAPRAIKEIRKFAEREMGTTDVRIDTRLNKHIWSKGIRSTPFRIRVRLARRRNHHHDSPNKLYTYVTYVPVSTFKNLQTENVESHHH

[0228] SEQ ID NO: 49MTKTKGEKINKSAHQHQHTRECTIHLAKRVQHQGFKKRAPRAIKEIRKFAEREMGTTDVRIDTRLNKHIWSKGIRSTPFRIRVRLARRRNDDEDSPNKLYTYVTYVPVSTFKNLQTENV ESHQH

[0229] SEQ ID NO: 50MTKTKGEKINKSADDDDKTRECTIHLAKRVHNIGFKKRAPRAIKEIRKFAEREMGTTDVRIDTRLNKHIWSKGIRSTPFRIRVRLARRRNDDEDSPNKLYTYVTYVPVSTFKNLQTENVESSDD

[0230] SEQ ID NO: 51MTKTKGEKINKSAEEEEETRECTIHLAKRVEEEGFKKRAPRAIKEIRKFAEREMGTTDVRIDTRLNKHIWSKGIRSTPFRIRVRLARRRNDDEDSPNKLYTYVTYVPVSTFKNLQTENVESEEE

[0231] SEQ ID NO: 52MTKTKGEKINKSARRRRRTRECTIHLAKRVRRRGFKKRAPRAIKEIRKFAEREMGTTDVR1DTRLNKH1WSKG1RSTPFR1RVRLARRRNDDEDSPNKLYTYVTYVPVSTFKNLQTENVESRRR

[0232] SEQ ID NO: 53MTKTKGEKINKSACCCCVTRECTIHLAKRVHNIGFKKRAPRAIKEIRKFAEREMGTTDVRIDTRLNKHIWSKGIRSTPFRIRVRLARRRNDDEDSPNKLYTYVTYVPVSTFKNLQTENVECCCC

[0233] SEQ ID NO: 54MTKTKGEKINKSAINEVVTRECTIHLAKRVHNIGFKKRAPRAIKEIRKFAEREMGTTDVRIDTRLNKHIWSKGIRSTPFRIRVRLARRRNDDEDSPNKLYTYVTYVPVSTFKNLQTENVEEPEA

[0234] SEQ ID NO: 55MHKVKCSELRIKDKKELTKQLDELKNELHHHHVAKVTGGAPSKLSKIRVVRKAIARVYIVMHQKQKENLRKVFKNKKYKPLDLRKKKTRAIRKALSPRDANRKTLKEIRKRSVFPQRKFAVKA

[0235] SEQ ID NO: 56MHKVKCSELRIKDKKELTKQLDELKNELQHQHVAKVTGGAPSKLSKIRVVRKAIARVYIVMHQKQKENLRKVFKNKKYKPLDLRKKKTRAIRKALSPRDANRKTLKEIRKRSVFPQRKFAVKA

[0236] SEQ ID NO: 57MEKVKCSELRIKDKKELTKQLDELKNELEEEEVAKVTGGAPSKLSKIRVVRKAIARVYIVMHQKQKENLRKVFKNKKYKPLDLRKKKTRAIRKALSPRDANRKTLKEIRKRSVFPQRK FAVKA

[0237] SEQ ID NO: 58MRKVKCSELRIKDKKELTKQLDELKNELRRRRVAKVTGGAPSKLSKIRVVRKAIARVYIVMHQKQKENLRKVFKNKKYKPLDLRKKKTRAIRKALSPRDANRKTLKEIRKRSVFPQRKFAVKA

[0238] SEQ ID NO: 59MVKVKCSELRIKDKKELTKQLDELKNELCCCCVAKVTGGAPSKLSKIRVVRKAIARVYIVMHQKQKENLRKVFKNKKYKPLDLRKKKTRAIRKALSPRDANRKTLKEIRKRSVFPQRKFAVKA

[0239] SEQ ID NO: 60MXXXHHHHRNKKQRKQQVKLRKPGFAVAKYVRMSPRKVRLVVDV1RGKSVQDAEDLLRFIPRSASEPVAKVLNSAKANAHHNDEHHHHRLFVKEAYVDAGPTLKRLIPRARGSANIIKKRTSHITIIVAEHHHH

[0240] SEQ ID NO: 61MXXXHQHQRNKKQRKQQVKLRKPGFAVAKYVRMSPRKVRLVVDVIRGKSVQDAEDLLRFIPRSASEPVAKVLNSAKANALHNDEHQHQRLFVKEAYVDAGPTLKRLIPRARGSANIIKKRTSHITIIVAEHQHQ

[0241] SEQ ID NO: 62MXXXEEEERNKKQRKQQVKLRKPGFAVAKYVRMSPRKVRLVVDVIRGKSVQDAEDLLRFIPRSASEPVAKVLNSAKANALHNDEEEEERLFVKEAYVDAGPTLKRLIPRARGSANIIKKRTSHITIIVAEEEEE

[0242] SEQ ID NO: 63MXXXRRRRRNKKQRKQQVKLRKPGFAVAKYVRMSPRKVRLVVDVIRGKSVQDAEDLLRFIPRSASEPVAKVLNSAKANALHNDERRRRRLFVKEAYVDAGPTLKRLIPRARGSANIIKKRTSHITIIVAERRRR

[0243] SEQ ID NO: 64MXXXCCCCRNKKQRKQQVKLRKPGFAVAKYVRMSPRKVRLVVDVIRGKSVQDAEDLLRFIPRSASEPVAKVLNSAKANALHNDECCCCRLFVKEAYVDAGPTLKRLIPRARGSANIIKKRTSHITIIVAECCCC

[0244] SEQ ID NO: 65MXXXEQTFRNKKQRKQQVKLRKPGFAVAKYVRMSPRKVRLVVDVIRGKSVQDAEDLLRFIPRSASEPVAKVLNSAKANALHNDEMLEDRLFVKEAYVDAGPTLKRLIPRARGSANIIKKRTSHITIIVAEEPEA

[0245] SEQ ID NO: 66MSHYDILQAPVISEKAYSAMERGVYSFWVSPKATKTEIKDAIQQHFHVRVIGISTMNVPGKRKRVGRFIGQRNDRKKAIVRLAHHQHIHHLAGQ

[0246] SEQ ID NO: 67MSHYDILQAPVISEKAYSAMERGVYSFWVSPKATKTEIKDAIQQHFQVRVIGISTMNVPGKRKRVGRFIGQRNDRKKAIVRLAEHQHIQHLAGQ

[0247] SEQ ID NO: 68MSHYDILQAPVISEKAYSAMERGVYSFWVSPKATKTEIKDAIQQEFEVRVIGISTMNVPGKRKRVGRF1GQRNDRKKA1VRLAEEQE1EELAGQ

[0248] SEQ ID NO: 69MSHYDILQAPVISEKAYSAMERGVYSFWVSPKATKTEIKDAIQQRFRVRVIGISTMNVPGKRKRVGRFIGQRNDRKKAIVRLARRQRIRRLAGQ

[0249] SEQ ID NO: 70MSHYDILQAPVISEKAYSAMERGVYSFWVSPKATKTEIKDAIQQCFCVRVIGISTMNVPGKRKRVGRFIGQRNDRKKAIVRLACCQCICCLAGQ

[0250] SEQ ID NO: 71MPRPSAGSHHNDKLHFKKGDHVIVLSGKHKGQHGKVLLALPRDQKVVVHGVNVITKNVKPSMTNPQGGQEQRELALHASKVALVDPETGKATRVRKQIVDGKKVRVAVASHHHH

[0251] SEQ ID NO: 72MPRPSAGSHHNDKLHFKKGDHVIVLSGKHKGQHGKVLLALPRDQKVVVQGVNVITKNVKPSMTNPQGGQEQRELALHASKVALVDPETGKATRVRKQIVDGKKVRVAVASHQHQ

[0252] SEQ ID NO: 73MPRPSAGSHHNDKLHFKKGDEVIVLSGKHKGQEGKVLLALPRDQKVVVEGVNVITKNVKPSMTNPQGGQEQRELALHASKVALVDPETGKATRVRKQIVDGKKVRVAVASEEEE

[0253] SEQ ID NO: 74MPRPSAGSHHNDKLHFKKGDRVIVLSGKHKGQRGKVLLALPRDQKVVVRGVNVITKNVKPSMTNPQGGQEQRELALHASKVALVDPETGKATRVRKQIVDGKKVRVAVASRRRR

[0254] SEQ ID NO: 75MPRPSAGSHHNDKLHFKKGDCVIVLSGKHKGQCGKVLLALPRDQKVVVCGVNVITKNVKPSMTNPQGGQEQRELALHASKVALVDPETGKATRVRKQIVDGKKVRVAVASCCCC

[0255] SEQ ID NO: 76MPRPSAGSHHNDKLHFKKGDTVIVLSGKHKGQTGKVLLALPRDQKVVVEGVNVITKNVKPSMTNPQGGQEQRELALHASKVALVDPETGKATRVRKQIVDGKKVRVAVASEPEA

[0256] SEQ ID NO: 77MKPSEMRHHQHTDFAKEIDAHKKELMELRFQAAAGQLAHPHHVRQLRREVAQLNTVK AELARHHHH

[0257] SEQ ID NO: 78MKPSEMRQHQHTDFAKEIDAQKKELMELRFQAAAGQLAQPHRVRQLRREVAQLNTVKAELARHQHQ

[0258] SEQ ID NO: 79MKPSEMREEQETDFAKEIDAEKKELMELRFQAAAGQLAEPHEVRQLRREVAQLNTVKAELAREEEE

[0259] SEQ ID NO: 80MKPSEMRRRQRTDFAKEIDARKKELMELRFQAAAGQLARPHRVRQLRREVAQLNTVKAELARRRRR

[0260] SEQ ID NO: 81MKPSEMRNLQATDFAKEIDARKKELMELRFQAAAGQLAQPHRVRQLRREVAQLNTVKAELARCCCC

[0261] SEQ ID NO: 82MKPSEMRNLQATDFAKEIDARKKELMELRFQAAAGQLAQPHRVRQLRREVAQLNTVKAELAREPEA

[0262] SEQ ID NO: 83MGISYSVEADHHHTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQHHHHHHMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0263] SEQ ID NO: 84MGISYSVEADHQHTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQHQHQHQMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0264] SEQ ID NO: 85MGISYSVEADPDTTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQDDDDKAMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0265] SEQ ID NO: 86MGISYSVEADEEETAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQEEEEEEMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0266] SEQ ID NO: 87MGISYSVEADRRRTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQRRRRRRMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVEL1LEEPEVED

[0267] SEQ ID NO: 88MGISYSVEADCCCTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQCCCCCCMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0268] SEQ ID NO: 89MGISYSVEADPDTTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADHQCCPGCCMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0269] SEQ ID NO: 90MGISYSVEADPDTTAKAMLRERQMSFKHSKAIAREIKGKTAGEAVDYLEAVIEGDQPVPFKQHNSGVGHKSKVDGWDAGRYPEKASKAFLDLLENAVGNADTQDPSRVGMTIKHVAAHKVGEQQGRKPRAMGRASAWNSPQVDVELILEEPEVED

[0270] SEQ ID NO: 91MSWDVIKHPHVTEKAMNDMDFQNKLQFAVDDRASKGEVADAVEEQYHHHVEQVNTQNTMDGEKKAVVRLSHHHDAQEVASRIGVF

[0271] SEQ ID NO: 92MSWDVIKHPHHQHKAMNDMDFQNKLQFAVDDRASKGEVADAVEEQYHQHVEQVNTQNTMDGEKKAVVRLSEDDDAQEVASRIGVF

[0272] SEQ ID NO: 93MSWDVIKHPHEEEKAMNDMDFQNKLQFAVDDRASKGEVADAVEEQYEEEVEQVNTQNTMDGEKKAVVRLSEDDDAQEVASRIGVF

[0273] SEQ ID NO: 94MSWDVIKHPHRRRKAMNDMDFQNKLQFAVDDRASKGEVADAVEEQYRRRVEQVNTQNTMDGEKKAVVRLSEDDDAQEVASRIGVF

[0274] SEQ ID NO: 95MSWDVIKHPHCCCKAMNDMDFQNKLQFAVDDRASKGEVADAVEEQYCCCVEQVNTQNTMDGEKKAVVRLSEDDDAQEVASRIGVF

[0275] SEQ ID NO: 96MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAHDTVEVLRGDFAHHEGEVINVDLDKAVIHVHHVTLHKTHHHEVPRPLDTSNVRVTDHHHHDEKREARLESEDDSA

[0276] SEQ ID NO: 97MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAHDTVEVLRGDFAHQEGEVINVDLDKAVIHVHQVTLEKTHQHEVPRPLDTSNVRVTDHQHQDEKREARLESEDDSA

[0277] SEQ ID NO: 98MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAGDTVEVLRGDFAGEEGEVINVDLDKAVIHVEDVTLEKTDGEEVPRPLDTSNVRVTDDDDKDEKREARLESE DDSA

[0278] SEQ ID NO: 99MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAEDTVEVLRGDFAEEEGEVINVDLDKAVIHVEEVTLEKTEEEEVPRPLDTSNVRVTDEEEEDEKREARLESEDDSA

[0279] SEQ ID NO: 100MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNARDTVEVLRGDFARREGEVINVDLDKAVIHVRRVTLEKTRRREVPRPLDTSNVRVTDRRRRDEKREARLESEDDSA

[0280] SEQ ID NO: 101MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAGDTVEVLRGDFAGEEGEVINVDLDKAVIHVEDVTLEKTCCCEVPRPLDTSNVRVTDCCCCDEKREARLESE DDSA

[0281] SEQ ID NO: 102MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAGDTVEVLRGDFAGEEGEVINVDLDKAVIHVEDVTLEKTDGEEVPRPLDTSNVRVTDYKDDDDKREARLESE DDSA

[0282] SEQ ID NO: 103MSKQPDKQRKSQRRAPLHERHKQVRATLSADLREEYGQRNVRVNAGDTVEVLRGDFAGEEGEVINVDLDKAVIHVEDVTLEKTDGEEVPRPLDTSNVRVTDLYDDDDKREARLESE DDSA

[0283] SEQ ID NO: 104MTVLHVQEIRDMTPAEREAELDDLKTELLNARAVQAAGHHHHHPGRIKELRKAIARIKT1QGEEGDLQENE

[0284] SEQ ID NO: 105MTVLHVQEIRDMTPAEREAELDDLKTELLNARAVQAAGHQHQHPGRIKELRKAIARIKTIQGEEGDLQENE

[0285] SEQ ID NO: 106MTVLHVQEIRDMTPAEREAELDDLKTELLNARAVQAAGDDDDKPGRIKELRKAIARIKTIQGEEGDLQENE

[0286] SEQ ID NO: 107MTVLHVQEIRDMTPAEREAELDDLKTELLNARAVQAAGEEEEEPGRIKELRKAIARIKTIQGEEGDLQENE

[0287] SEQ ID NO: 108MTVLHVQEIRDMTPAEREAELDDLKTELLNARAVQAAGRRRRRPGRIKELRKAIARIKTIQGEEGDLQENE

[0288] SEQ ID NO: 109MTVLHVQEIRDMTPAEREAELDDLKTELLNARAVQAAGCCCCCPGRIKELRKAIARIKTIQGEEGDLQENE

[0289] SEQ ID NO: 110MSHHHHEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFHHHHHHIVEAETAE

[0290] SEQ ID NO: 111MSASDFEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFHQHQHQIVEAETAE

[0291] SEQ ID NO: 112MSASDFEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFDDDDKAIVEAETAE

[0292] SEQ ID NO: 113MSASDFEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFEEEEEAIVEAETAE

[0293] SEQ ID NO: 114MSASDFEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFRRRRRRIVEAETAE

[0294] SEQ ID NO: 115MSASDFEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFCCCCCCIVEAETAE

[0295] SEQ ID NO: 116MSASDFEERVVTIPLRDARAEPNHKRADKAMILIREHLAKHFSVDEDAVRLDPSINEAAWARGRANTPSKIRVRAARFCCPGCCIVEAETAE

[0296] SEQ ID NO: 117MVRYSLDPHHPHHSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCV PFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKHLHVHHLVIE HIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVAQKKKISQKKL KKQKLMARE

[0297] SEQ ID NO: 118MVRYSLDPHQPHQSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCV PFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKHLQVHQLVIE HIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVAQKKKISQKKL KKQKLMARE

[0298] SEQ ID NO: 119MVRYSLDPEEPEESCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVP FRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKELEVEELVIEHI QVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVAQKKKISQKKLKK QKLMARE

[0299] SEQ ID NO: 120MVRYSLDPRRPRRSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKRLRVRRLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVAQKKKISQKKLKKQKLMARESEQ ID NO: 121MVRYSLDPCCPCCSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKCLCVCCLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKEQIVPKPEEEVAQKKKISQKKLKKQKLMARE

[0300] SEQ ID NO: 122MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFHHHHHHEEEEDED

[0301] SEQ ID NO: 123MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFHQHQHQEEEEDED

[0302] SEQ ID NO: 124MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFDDDDKEEEEEDED

[0303] SEQ ID NO: 125MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFEEEEEEEEEEDED

[0304] SEQ ID NO: 126MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFRRRRRREEEEDED

[0305] SEQ ID NO: 127MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFCCCCCCEEEEDED

[0306] SEQ ID NO: 128MAPVKKLVVKGGKKKKQVLKFTLDCTHPVEDGIMDAANFEQFLQERIKVNGKAGNLGGGVVTIERSKSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFCCPGCCEEEEDED

[0307] SEQ ID NO: 129MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQREHHHHHHVHVGIHPSKVVITRLKLDKDRKK 1LERKAKSRQVGKEKGKYKEET1EKMQE

[0308] SEQ ID NO: 130MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQREHQHQHQVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0309] SEQ ID NO: 131MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQREDDDDKTVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0310] SEQ ID NO: 132MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQREEEEEEEVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0311] SEQ ID NO: 133MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQRERRRRRRVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0312] SEQ ID NO: 134MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQRECCCCCCVHVGIHPSKVVITRLKLDKDRKKI LERKAKSRQVGKEKGKYKEETIEKMQE

[0313] SEQ ID NO: 135MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGQQIGKVVQVYRKKYVIYIERVQRECCPGCCVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0314] SEQ ID NO: 136MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKHHHHHHSPNKLYTLVTYVPVTTFKNLQTVNVDEN

[0315] SEQ ID NO: 137MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVR1DTRLNKAVWAKG1RNVPYR1RVRLSRKHQHQHQSPNKLYTLVTYVPVTTFKNLQTVNVDEN

[0316] SEQ ID NO: 138MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKDDDDKDSPNKLYTLVTYVPVTTFKNLQTVNVDEN

[0317] SEQ ID NO: 139MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKEEEEEESPNKLYTLVTYVPVTTFKNLQTVNVDEN

[0318] SEQ ID NO: 140MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKRRRRRRSPNKLYTLVTYVPVTTFKNLQTVNVDEN

[0319] SEQ ID NO: 141MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKCCCCCCSPNKLYTLVTYVPVTTFKNLQTVNVDEN

[0320] SEQ ID NO: 142MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKCCPGCCSPNKLYTLVTYVPVTTFKNLQT VNVDEN

[0321] SEQ ID NO: 143MHHHKARDLRGKKKEELLKQLDDLKVELSQLRVAKVTHHHASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKYAVKA

[0322] SEQ ID NO: 144MHQHKARDLRGKKKEELLKQLDDLKVELSQLRVAKVTHQHASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKYAVKA

[0323] SEQ ID NO: 145MEEEKARDLRGKKKEELLKQLDDLKVELSQLRVAKVTEEEASKLSK1RVVRKS1ARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKYAVKA

[0324] SEQ ID NO: 146MRRRKARDLRGKKKEELLKQLDDLKVELSQLRVAKVTRRRASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKYAVKA

[0325] SEQ ID NO: 147MCCCKARDLRGKKKEELLKQLDDLKVELSQLRVAKVTCCCASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKYAVKA

[0326] SEQ ID NO: 148MARYGATSTNPHKSASARGSYLRVSYKNTRETAQAISGWNLQKAQKYLDQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAASNAEAKGLHAHRLYVSHIQVNQAPKQRRRTFRAHGRINKYESSPSHIELVVTEKEHHVEHASEKKVARLSSRQRGRLATQKRITA

[0327] SEQ ID NO: 149MARYGATSTNPHKSASARGSYLRVSYKNTRETAQAISGWNLQKAQKYLDQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAASNAEAKGLHAQRLYVSHIQVNQAPKQRRRTFRAHGRINKYESSPSHIELVVTEKEHQVEHASEKKVARLSSRQRGRLATQKRITA

[0328] SEQ ID NO: 150MARYGATSTNPEKSASARGSYLRVSYKNTRETAQAISGWNLQKAQKYLDQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAASNAEAKGLEAERLYVSHIQVNQAPKQRRRTFRAHGRINKYESSPSHIELVVTEKEEEVEEASEKKVARLSSRQRGRLATQKRITA

[0329] SEQ ID NO: 151MANLRTQKRLAASVAGVGKRKIWMDPHQTAEIAQAHSRNAIRKLVKHQTIVKKPTRIHSRSRTRAMAESKRNGRHTGYGKRKGTKEARLPSQVVWIRRLRVLRRLLAKYRDAGKIDRHLYHSLYKSAKGNTFKHKRALVEHIIQAKADAQREKALKEEAEARRSKNRAARERRAQRVAEKREALLKEDA

[0330] SEQ ID NO: 152MANLRTQKRLAASVAGVGKRKIWMDPHQTAEIAQAHSRNAIRKLVKHQTIVKKPTRIHSRSRTRAMAESKRNGRHTGYGKRKGTKEARLPSQVVWIRRLRVLRRLLAKYRDAGKIDRHLYHSLYKSAKGNTFKHKRALVEHIIQAKADAQREKALKEEAEARRSKNRAARERRAQRVAEKREALLKEDA

[0331] SEQ ID NO: 153MANLRTQKRLAASVAGVGKRKIWMDPEETAEIAQAESRNAIRKLVKEETIVKKPTRIHSRSRTRAMAESKRNGRHTGYGKRKGTKEARLPSQVVWIRRLRVLRRLLAKYRDAGKIDRHLYHSLYKSAKGNTFKHKRALVEHIIQAKADAQREKALKEEAEARRSKNRAARERRAQRVAEKREALLKEDA

[0332] SEQ ID NO: 154MAPSTKATAAKKAVVKGTNGKKALKVRTSASFRLPKTLKLARSPKYATKAVPHYNRLDSYKVIEQPITSETAMKKVEDGNTLVFKVSLKANKYQIKKAVKELYHHHVLSVNTLVRPNGTKKAYVRLTHHHDALDIANRHHHH

[0333] SEQ ID NO: 155MAPSTKATAAKKAVVKGTNGKKALKVRTSASFRLPKTLKLARSPKYATKAVPHYNRLDSYKVIEQPITSETAMKKVEDGNTLVFKVSLKANKYQIKKAVKELYHQHVLSVNTLVRPNGTKKAYVRLTHQHDALDIANRHQHQ

[0334] SEQ ID NO: 156MAPSTKATAAKKAVVKGTNGKKALKVRTSASFRLPKTLKLARSPKYATKAVPHYNRLDSYKVIEQPITSETAMKKVEDGNTLVFKVSLKANKYQIKKAVKELYEEEVLSVNTLVRPNGTKKAYVRLTEEEDALDIANREEEE

[0335] SEQ ID NO: 157MAPSTKATAAKKAVVKGTNGKKALKVRTSASFRLPKTLKLARSPKYATKAVPHYNRLDSYKVIEQPITSETAMKKVEDGNTLVFKVSLKANKYQIKKAVKELYRRRVLSVNTLVRPNGTKKAYVRLTRRRDALDIANRRRRR

[0336] SEQ ID NO: 158MAPSTKATAAKKAVVKGTNGKKALKVRTSASFRLPKTLKLARSPKYATKAVPHYNRLDSYKVIEQPITSETAMKKVEDGNTLVFKVSLKANKYQIKKAVKELYCCCVLSVNTLVRPNGTKKAYVRLTCCCDALDIANRCCCC

[0337] SEQ ID NO: 159MAPSTKATAAKKAVVKGTNGKKALKVRTSASFRLPKTLKLARSPKYATKAVPHYNRLDSYKVIEQPITSETAMKKVEDGNTLVFKVSLKANKYQIKKAVKELYEVDVLSVNTLVRPNGTKKAYVRLTADFDALDIANREPEA

[0338] SEQ ID NO: 160MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKHHHHHHVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGHHHH

[0339] SEQ ID NO: 161MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKHQHQHQVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGHQHQ

[0340] SEQ ID NO: 162MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKKDDDDKVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGGKLE

[0341] SEQ ID NO: 163MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKEEEEEEVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGEEEE

[0342] SEQ ID NO: 164MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKRRRRRRVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGRRRR

[0343] SEQ ID NO: 165MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKCCCCCCVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGCCCC

[0344] SEQ ID NO: 166MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKKGQEGKVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGEPEA

[0345] SEQ ID NO: 167MAKQSLDVSSDRRKARKAYFTAPSSERRVLLSAPLSKELREQYNIKALPIRKEDEVLVVRGSKCCPGCCVSSVYRLKFAVQVDKLTKEKSNGASVPTNIHPSKVVITKLHLDKDRKALIQRKGGKLE

[0346] SEQ ID NO: 168MAGLHHHHHREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRHNHEEDAKNPLFSYVEPVVVASAKGLHTHHHHHHHH

[0347] SEQ ID NO: 169MAGLHQHQHREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRHNQEEDAKNPLFSYVEPVVVASAKGLHTHQHQHQHQ

[0348] SEQ ID NO: 170MAGLDDDDKREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTVVVDDDDK

[0349] SEQ ID NO: 171MAGLEEEEEREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTEEEEEEEE

[0350] SEQ ID NO: 172MAGLRRRRRREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTRRRRRRRR

[0351] SEQ ID NO: 173MAGLCCCCCREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTCCCCCCCC

[0352] SEQ ID NO: 174MAGLKDVVTREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTDYKDDDDK

[0353] SEQ ID NO: 175MAGLKDVVTREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTVVVDEPEA

[0354] SEQ ID NO: 176MAGLKDVVTREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTCCPGCCEA

[0355] SEQ ID NO: 177MAGLKDVVTREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTDLYDDDDK

[0356] SEQ ID NO: 178MAGLKDVVTREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTTQDPSRVG

[0357] SEQ ID NO: 179MAGLKDVVTREYTINMHKRLHGVSFKKRAPKAVKEIKKFAKLHMGTDDVRLDPKLNQEIWKRGIKGVPFRLRLRISRRRNEEEDAKNPLFSYVEPVVVASAKGLHTWSHPQFEK

[0358] SEQ ID NO: 180MAHHKAYHLRHHHKDQLEQQLVELKKELAELKVQKLHHHHLPKINTVRKNIARVLTVISQNQRQAVRELYKGKKYQPKDLRAKKTRALRRALTKFEASQVTEKQRKKQIAFPQRKYAIKA

[0359] SEQ ID NO: 181MAHQKAYHLRHHHKDQLEQQLVELKKELAELKVQKLHHHHLPKINTVRKNIARVLTVISQNQRQAVRELYKGKKYQPKDLRAKKTRALRRALTKFEASQVTEKQRKKQIAFPQRKY AIKA

[0360] SEQ ID NO: 182MAEEKAYELREEEKDQLEQQLVELKKELAELKVQKLEEEELPKINTVRKNIARVLTVISQNQRQAVRELYKGKKYQPKDLRAKKTRALRRALTKFEASQVTEKQRKKQIAFPQRKYAIKA

[0361] SEQ ID NO: 183MARRKAYRLRRRRKDQLEQQLVELKKELAELKVQKLRRRRLPKINTVRKNIARVLTVISQNQRQAVRELYKGKKYQPKDLRAKKTRALRRALTKFEASQVTEKQRKKQIAFPQRKYA IKA

[0362] SEQ ID NO: 184MACCKAYCLRCCCKDQLEQQLVELKKELAELKVQKLCRCCLPKINTVRKNIARVLTVISQNQRQAVRELYKGKKYQPKDLRAKKTRALRRALTKFEASQVTEKQRKKQIAFPQRKYA IKA

[0363] SEQ ID NO: 185MVRYSLDPENPHHSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLKKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKHLHVDSLV1EHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTHHHHIVPKPEEEVAQKKKISQKKL KKQKLMARE

[0364] SEQ ID NO: 186MVRYSLDPENPHQSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLKKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKHLQVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTHQHQIVPKPEEEVAQKKKISQKKL KKQKLMARE

[0365] SEQ ID NO: 187MVRYSLDPENPEESCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLKKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKELEVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEEEEIVPKPEEEVAQKKKISQKKLKK QKLMARE

[0366] SEQ ID NO: 188MVRYSLDPENPRRSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLKKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKRLRVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTRRRRIVPKPEEEVAQKKKISQKKLK KQKLMARE

[0367] SEQ ID NO: 189MVRYSLDPENPCCSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLKKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKCLCVDSLVIEH IQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTCCCCIVPKPEEEVAQKKKISQKKLKKQKLMARE

[0368] SEQ ID NO: 190MSMLRLQKRLASSVLRCGKKKVWLDPNHTNEIAHHNSRQQIRKLIKDGHIIRKPVTVHSRARCRKNTLARRKGRHMGIGKRKGTANARMPEKVTWMRRMRILRRLLRRYRESKKIDRHMYHSLYLKVKGNVFKNKRILMEHIHKLKADKARKKLLADQAEARRSKTKEARKRREERLQ AKKEEIIKTLS KEEETKK

[0369] SEQ ID NO: 191MSMLRLQKRLASSVLRCGKKKVWLDPNHTNEIAQHNSRQQIRKLIKDGQIIRKPVTVHSRARCRKNTLARRKGRHMGIGKRKGTANARMPEKVTWMRRMRILRRLLRRYRESKKIDRHMYHSLYLKVKGNVFKNKR1LMEH1HKLKADKARKKLLADQAEARRSKTKEARKRREERLQ AKKEEIIKTLS KEEETKK

[0370] SEQ ID NO: 192MSMLRLQKRLASSVLRCGKKKVWLDPNETNEIAEENSRQQIRKLIKDGEIIRKPVTVHSRARCRKNTLARRKGRHMGIGKRKGTANARMPEKVTWMRRMRILRRLLRRYRESKKIDRHMYHSLYLKVKGNVFKNKRILMEHIHKLKADKARKKLLADQAEARRSKTKEARKRREERLQAKKEEIIKTLSKEEETKK

[0371] SEQ ID NO: 193MSMLRLQKRLASSVLRCGKKKVWLDPNRTNEIARRNSRQQIRKLIKDGRIIRKPVTVHSRARCRKNTLARRKGRHMGIGKRKGTANARMPEKVTWMRRMRILRRLLRRYRESKKIDRHMYHSLYLKVKGNVFKNKRILMEHIHKLKADKARKKLLADQAEARRSKTKEARKRREERLQ AKKEEIIKTLS KEEETKK

[0372] SEQ ID NO: 194MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYHHHVAKVNTLIRPDGEKKAYVRLAHHHDALDVANKIGII

[0373] SEQ ID NO: 195MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYHQHVAKVNTLIRPDGEKKAYVRLAHQHDALDVANKIGII

[0374] SEQ ID NO: 196MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYEEEVAKVNTLIRPDGEKKAYVRLAEEEDALDVANKIGII

[0375] SEQ ID NO: 197MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYRRRVAKVNTLIRPDGEKKAYVRLARRRDALDVANKIGII

[0376] SEQ ID NO: 198MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKHDHVQVVRGHYKHHHHGKVVQVYRKKYVIYIHHVQRHKANHTTVHVGIHPSKVVITHLHLDKDRKK1LERKAKSRQVGKEKGKYKEET1EKMQE

[0377] SEQ ID NO: 199MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKHDQVQVVRGHYKHQHQGKVVQVYRKKYVIYIHQVQRHKANQTTVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0378] SEQ ID NO: 200MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKEDEVQVVRGHYKEEEEGKVVQVYRKKYVIYIEEVQREKANETTVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0379] SEQ ID NO: 201MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKRDRVQVVRGHYKRRRRGKVVQVYRKKYVIYIRRVQRRKANRTTVHVGIHPSKVVITRLKLDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0380] SEQ ID NO: 202MAPAKKGGEKKKGRSAHHHHVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKHNHHEHSPNHLYTLHTHVPVTTFKNLQTVHHHHH

[0381] SEQ ID NO: 203MAPAKKGGEKKKGRSAHQHQVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKHNQHEHSPNKLYTLVTYVPVTTFKNLQTVHQHQH

[0382] SEQ ID NO: 204MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKRNEDEDSPNKLYTLVTYVPVTTFKNLQTVDDDDK

[0383] SEQ ID NO: 205MAPAKKGGEKKKGRSAEEEEVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKENEEEESPNKLYTLVTYVPVTTFKNLQTVEEEEE

[0384] SEQ ID NO: 206MAPAKKGGEKKKGRSARRRRVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVR1DTRLNKAVWAKG1RNVPYR1RVRLSRKRNRRERSPNKLYTLVTYVPVTTFKNLQTVRRRRR

[0385] SEQ ID NO: 207MAPAKKGGEKKKGRSACCCCVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKCNCCECSPNKLYTLVTYVPVTTFKNLQTVCCCCC

[0386] SEQ ID NO: 208MAPAKKGGEKKKGRSAINEVVTREYTINIHKRIHGVGFKKRAPRALKEIRKFAMKEMGTPDVRIDTRLNKAVWAKGIRNVPYRIRVRLSRKRNEDEDSPNKLYTLVTYVPVTTFKNLQTVNEPEA

[0387] SEQ ID NO: 209MHHHHARDLRHHHKEELLKQLDDLKVELSQLRVAKVHHHHASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLTKHEEKLKTKKQQRKEREYPLRKYAVKA

[0388] SEQ ID NO: 210MHQHQARDLRHQHKEELLKQLDDLKVELSQLRVAKVHQHQASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLTKHEEKLKTKKQQRKERLYPLRKYAVKA

[0389] SEQ ID NO: 211MEEEEARDLREEEKEELLKQLDDLKVELSQLRVAKVEEEEASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLTKHEEKLKTKKQQRKERLYPLRK YAVKA

[0390] SEQ ID NO: 212MRRRRARDLRRRRKEELLKQLDDLKVELSQLRVAKVRRRRASKLSKIRVVRKSIARVLTVINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLTKHEEKLKTKKQQRKERLYPLR KYAVKA

[0391] SEQ ID NO: 213MCCCCARDLRCCCKEELLKQLDDLKVELSQLRVAKVCCCCASKLSKIRVVRKSIARVLT VINQTQKENLRKFYKGKKYKPLDLRPKKTRAMRRRLTKHEEKLKTKKQQRKERLYPLR KYAVKA

[0392] SEQ ID NO: 214MVKYSKEPDNPHHSCKARGSSLRVHFKNTRETAHALRKMPLDKAKRYLEDVLAHKQA1 PFTRFCRGVGRTAQAKNRHSSGQGRWPVKSTKFILDLLKNAESNADVKHLHVDHLFISH IQVNQAQKQRRRTYRAHGRINPYMSHPCHIELTLSHHHHYVKKEPETQLAPRKTKGSS

[0393] SEQ ID NO: 215MVKYSKEPDNPHQSCKARGSSLRVHFKNTRETAHALRKMPLDKAKRYLEDVLAHKQAI PFTRFCRGVGRTAQAKNRHSSGQGRWPVKSTKFILDLLKNAESNADVKHLQVDHLFISH IQVNQAQKQRRRTYRAHGRINPYMSHPCHIELTLSHQHQYVKKEPETQLAPRKTKGSS

[0394] SEQ ID NO: 216MVKYSKEPDNPEESCKARGSSLRVHFKNTRETAHALRKMPLDKAKRYLEDVLAHKQAI PFTRFCRGVGRTAQAKNRHSSGQGRWPVKSTKFILDLLKNAESNADVKELEVDELFISHI QVNQAQKQRRRTYRAHGRINPYMSHPCHIELTLSEEEEYVKKEPETQLAPRKTKGSS

[0395] SEQ ID NO: 217MVKYSKEPDNPRRSCKARGSSLRVHFKNTRETAHALRKMPLDKAKRYLEDVLAHKQAI PFTRFCRGVGRTAQAKNRHSSGQGRWPVKSTKFILDLLKNAESNADVKRLRVDRLFISHI QVNQAQKQRRRTYRAHGRINPYMSHPCHIELTLSRRRRYVKKEPETQLAPRKTKGSS

[0396] SEQ ID NO: 218MVKYSKEPDNPCCSCKARGSSLRVHFKNTRETAHALRKMPLDKAKRYLEDVLAHKQAI PFTRFCRGVGRTAQAKNRHSSGQGRWPVKSTKFILDLLKNAESNADVKCLCVDCLFISHI QVNQAQKQRRRTYRAHGRINPYMSHPCHIELTLSCCCCYVKKEPETQLAPRKTKGSS

[0397] SEQ ID NO: 219MVKYSKEPDNPTKSCKARGSSLRVHFKNTRETAHALRKMPLDKAKRYLEDVLAHKQAI PFTRFCRGVGRTAQAKNRHSSGQGRWPVKSTKFILDLLKNAESNADVKGLDVDLLFISHIQVNQAQKQRRRTYRAHGRINPYMSHPCHIELTLSEPEAYVKKEPETQLAPRKTKGSS

[0398] SEQ ID NO: 220MSKGSSGGGAMKGGKKKGATFVIDCSKPVEDKIMEIASLEKFLQERIKVGGKAGALHHSVTVTRHHHKITVTSHTTFSKRYLKYLTKKYLKKNNVRDWLRVISSNKDRNVYELRYFNIAENEAEEED

[0399] SEQ ID NO: 221MSKGHQHGGAMKGGKKKGATFVIDCSKPVEDKIMEIASLEKFLQERIKVGGKAGALHQSVTVTRDKTKITVTSHTTFSKRYLKYLTKKYLKKNNVRDWLRVISSNKDRNVYELRYFN IAENEAEEED

[0400] SEQ ID NO: 222MSKGEEEGGAMKGGKKKGATFVIDCSKPVEDKIMEIASLEKFLQERIKVGGKAGALEESVTVTRDKTKITVTSETTFSKRYLKYLTKKYLKKNNVRDWLRVISSNKDRNVYELRYFNIAENEAEEED

[0401] SEQ ID NO: 223MSKGRRRGGAMKGGKKKGATFVIDCSKPVEDKIMEIASLEKFLQERIKVGGKAGALRRSVTVTRDKTKITVTSRTTFSKRYLKYLTKKYLKKNNVRDWLRVISSNKDRNVYELRYFNIAENEAEEED

[0402] SEQ ID NO: 224MSKGCCCGGAMKGGKKKGATFVIDCSKPVEDKIMEIASLEKFLQERIKVGGKAGALCCSVTVTRDKTKITVTSCTTFSKRYLKYLTKKYLKKNNVRDWLRVISSNKDRNVYELRYFNIAENEAEEED

[0403] SEQ ID NO: 225MKYNPRVSSSRRKSRKAHFTAPSSVRRVLMSAPLSSDLRTKYNVRSMPVRKHDHVQVVRGTYKHRHGKVVQVYRKKWVIHIHHITREKVNHSTVNVGIHPSKVVVTHLRHDKDRKSLLDRKAKGRAHHHHDKGTKFTAEDIMQTVD

[0404] SEQ ID NO: 226MKYNPRVSSSRRKSRKAHFTAPSSVRRVLMSAPLSSDLRTKYNVRSMPVRKHDQVQVVRGTYKHRQGKVVQVYRKKWVIHIHQITREKVNHSTVNVGIHPSKVVVTHLRQDKDRKSLLDRKAKGRAHQHQDKGTKFTAEDIMQTVD

[0405] SEQ ID NO: 227MKYNPRVSSSRRKSRKAHFTAPSSVRRVLMSAPLSSDLRTKYNVRSMPVRKEDEVQVVRGTYKEREGKVVQVYRKKWVIHIEEITREKVNESTVNVGIHPSKVVVTELREDKDRKSLLDRKAKGRAEEEEDKGTKFTAEDIMQTVD

[0406] SEQ ID NO: 228MKYNPRVSSSRRKSRKAHFTAPSSVRRVLMSAPLSSDLRTKYNVRSMPVRKRDRVQVVRGTYKRRRGKVVQVYRKKWVIHIRRITREKVNRSTVNVGIHPSKVVVTRLRRDKDRKSLLDRKAKGRARRRRDKGTKFTAEDIMQTVD

[0407] SEQ ID NO: 229MKYNPRVSSSRRKSRKAHFTAPSSVRRVLMSAPLSSDLRTKYNVRSMPVRKCDCVQVVRGTYKCRCGKVVQVYRKKWVIHICCITREKVNCSTVNVGIHPSKVVVTCLRCDKDRKSLLDRKAKGRACCCCDKGTKFTAED1MQTVD

[0408] SEQ ID NO: 230MKYNPRVSSSRRKSRKAHFTAPSSVRRVLMSAPLSSDLRTKYNVRSMPVRKDDEVQVVRGTYKGREGKVVQVYRKKWVIHIERITREKVNGSTVNVGIHPSKVVVTKLRLDKDRKSLLDRKAKGRAEPEADKGTKFTAEDIMQTVD

[0409] SEQ ID NO: 231MSDKTKGRKEHHHHREYTINLHKRLHSCTFKKKAPTAIKEIRKFAQKAMGTKDVRVDVKLNKQIWSRGIRSVPRRVRVRIARKRNHDHHAKEHLYSLVTVAEIPAEGLKGLGTKIIHHH H

[0410] SEQ ID NO: 232MSDKTKGRKEHQHQREYTINLHKRLHSCTFKKKAPTAIKEIRKFAQKAMGTKDVRVDVKLNKQIWSRGIRSVPRRVRVRIARKRNHDQHAKEELYSLVTVAEIPAEGLKGLGTKIIHQH Q

[0411] SEQ ID NO: 233MSDKTKGRKEEEEEREYTINLHKRLESEEFKKKAPTAIKEIRKFAQKAMGTKDVRVDVKLNKQIWSRGIRSVPRRVRVRIARKRNDDEDAKEELYSLVTVAEIPAEGLKGLGTKIIEEEE

[0412] SEQ ID NO: 234MSDKTKGRKERRRRREYTINLHKRLHSCTFKKKAPTAIKEIRKFAQKAMGTKDVRVDVKLNKQIWSRGIRSVPRRVRVRIARKRNRDRRAKEELYSLVTVAEIPAEGLKGLGTKIIRRR R

[0413] SEQ ID NO: 235MSDKTKGRKECCCCREYTINLHKRLHSCTFKKKAPTAIKEIRKFAQKAMGTKDVRVDVKLNKQIWSRGIRSVPRRVRVRIARKRNCDCCAKEELYSLVTVAEIPAEGLKGLGTKIICCC C

[0414] SEQ ID NO: 236MSDKTKGRKEEVVTREYTINLHKRLHSCTFKKKAPTAIKEIRKFAQKAMGTKDVRVDVKLNKQIWSRGIRSVPRRVRVRIARKRNDDEDAKEELYSLVTVAEIPAEGLKGLGTKIIEPE A

[0415] SEQ ID NO: 237MHRIHVHELRHHHKTELLAQLKDLKAELALLRVAKVTGGAPNKLSKIKVVRLSIAQVLTVISQKQKTALREAYKNKKYLPLDLRPKKTRAIRKRLTKHQVSLKTEREKKKEMYFPIRK YAIKV

[0416] SEQ ID NO: 238MHRIQVHELRHQHKTELLAQLKDLKAELALLRVAKVTGGAPNKLSKIKVVRLSIAQVLTVISQKQKTALREAYKNKKYLPLDLRPKKTRAIRKRLTKHQVSLKTEREKKKEMYFPIRKYAIKV

[0417] SEQ ID NO: 239MERIEVHELREEEKTELLAQLKDLKAELALLRVAKVTGGAPNKLSKIKVVRLSIAQVLTVISQKQKTALREAYKNKKYLPLDLRPKKTRAIRKRLTKHQVSLKTEREKKKEMYFPIRKY AIKV

[0418] SEQ ID NO: 240MRRIRVHELRRRRKTELLAQLKDLKAELALLRVAKVTGGAPNKLSKIKVVRLSIAQVLTVISQKQKTALREAYKNKKYLPLDLRPKKTRAIRKRLTKHQVSLKTEREKKKEMYFPIRKYAIKV

[0419] SEQ ID NO: 241MCRICVHELRCCCKTELLAQLKDLKAELALLRVAKVTGGAPNKLSKIKVVRLSIAQVLTVISQKQKTALREAYKNKKYLPLDLRPKKTRAIRKRLTKHQVSLKTEREKKKEMYFPIRK YAIKV

[0420] SEQ ID NO: 242MPKQIHEIKDFLLTARRKDARTVKIKKNKDMVKFKVRCSKYLYTLCVSDFEKADKLKQSLHHHLHVQDL

[0421] SEQ ID NO: 243MPKQIHEIKDFLLTARRKDARTVKIKKNKDMVKFKVRCSKYLYTLCVSDFEKADKLKQSLHQHLQVQDL

[0422] SEQ ID NO: 244MPKQIHEIKDFLLTARRKDARTVKIKKNKDMVKFKVRCSKYLYTLCVSDFEKADKLKQSLEEELEVQDL

[0423] SEQ ID NO: 245MPKQIHEIKDFLLTARRKDARTVKIKKNKDMVKFKVRCSKYLYTLCVSDFEKADKLKQSLRRRLRVQDL

[0424] SEQ ID NO: 246MPKQIHEIKDFLLTARRKDARTVKIKKNKDMVKFKVRCSKYLYTLCVSDFEKADKLKQSLCCCLCVQDL

[0425] SEQ ID NO: 247MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGHQHGKVVQVYRKKYVIYIERVQREKANGTTVHVGIHPSKVVITHLHHDKDRKKILERKAKSRQVGKEKGK

[0426] SEQ ID NO: 248MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGHQHGKVVQVYRKKYVIYIERVQREKANGTTVHVGIHPSKVVITHLQHDKDRKKILERKAKSRQVGKEKGK

[0427] SEQ ID NO: 249MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGEQEGKVVQVYRKKYVIYIERVQREKANGTTVHVGIHPSKVVITELEEDKDRKKILERKAKSRQVGKEKGK

[0428] SEQ ID NO: 250MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGRQRGKVVQVYRKKYVIYIERVQREKANGTTVHVGIHPSKVVITRLRRDKDRKKILERKAKSRQVGKEKGK

[0429] SEQ ID NO: 251MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKDDEVQVVRGHYKGCQCGKVVQVYRKKYVIYIERVQREKANGTTVHVGIHPSKVVITCLCCDKDRKKILERKAKSRQVGKEKGK

[0430] SEQ ID NO: 252MAKIKARDLRHHHKEELLKQLEDLKVELSQLRVAKVTGGAASKLSKIRVVRKSIARVLTVINQTQKENLRHHYHGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKFAVKA

[0431] SEQ ID NO: 253MAKIKARDLRHQHKEELLKQLEDLKVELSQLRVAKVTGGAASKLSKIRVVRKSIARVLTVINQTQKENLRHQYHGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKFAVKA

[0432] SEQ ID NO: 254MAKIKARDLREEEKEELLKQLEDLKVELSQLRVAKVTGGAASKLSKIRVVRKSIARVLTV1NQTQKENLREEYEGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKFAVKA

[0433] SEQ ID NO: 255MAKIKARDLRRRRKEELLKQLEDLKVELSQLRVAKVTGGAASKLSKIRVVRKSIARVLTVINQTQKENLRRRYRGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKFAVKA

[0434] SEQ ID NO: 256MAKIKARDLRCCCKEELLKQLEDLKVELSQLRVAKVTGGAASKLSKIRVVRKSIARVLTVINQTQKENLRCCYCGKKYKPLDLRPKKTRAMRRRLNKHEENLKTKKQQRKERLYPLRKFAVKA

[0435] SEQ ID NO: 257MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSHHHLHVFLEKNKMYSREWNRDVQYRGRVRVQLKHHHHHHHHHHFPSRKSVMLYAAEMIPKLKT

[0436] SEQ ID NO: 258MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSHQHLQVFLEKNKMYSREWNRDVQYRGRVRVQLKHQHQHQHQHQFPSRKSVMLYAAEMIPKLKT

[0437] SEQ ID NO: 259MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSAVGLNVFLEKNKMYSREWNRDVQYRGRVRVQLKQEDDDDKLVQFPSRKSVMLYAAEMIPKLKT

[0438] SEQ ID NO: 260MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSEEELEVFLEKNKMYSREWNRDVQYRGRVRVQLKEEEEEEEEEEFPSRKSVMLYAAEMIPKLKT

[0439] SEQ ID NO: 261MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSRRRLRVFLEKNKMYSREWNRDVQYRGRVRVQLKRRRRRRRRRRFPSRKSVMLYAAEMIPKLKT

[0440] SEQ ID NO: 262MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSCCCLCVFLEKNKMYSREWNRDVQYRGRVRVQLKCCCCCCCCCCFPSRKSVMLYAAEMIPKLKT

[0441] SEQ ID NO: 263MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSAVGLNVFLEKNKMYSREWNRDVQYRGRVRVQLKQDYKDDDDKQFPSRKSVMLYAAEMIPKLKT

[0442] SEQ ID NO: 264MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSAVGLNVFLEKNKMYSREWNRDVQYRGRVRVQLKQECCPGCCVQFPSRKSVMLYAAEMIPKLKT

[0443] SEQ ID NO: 265MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSAVGLNVFLEKNKMYSREWNRDVQYRGRVRVQLKQDLYDDDDKQFPSRKSVMLYAAEMIPKLKT

[0444] SEQ ID NO: 266MXXXXXXXXXXXDRFICIYPAYLNNKKTIAEGRRIPISKAVENPTATEIQDVCSAVGLNVFLEKNKMYSREWNRDVQYRGRVRVQLKQWSHPQFEKQFPSRKSVMLYAAEMIPKLKT

[0445] SEQ ID NO: 267MXXADLGRKITSALRSLSNATIINEEVLNAMLKEVCTALLEADVNIKLVKQLRENVKSAIDLEEMASGLNKRKMIQHAVFKELVKLVDPGVKAWTPTKGKQNVIMFVGLQGSGKTTTCSKLAYYYQRKGWKTCLICADTFRAGAFDQLKQNATKARIPFYGSYHEHDPVIIASEGVEKFKNENFEIIIVDTSGRHHQHDSLFEEMLQVANAIQPDNIVYVMDASIGQACEAQAKAFKDKVDVASVIVTKLDGHAHGGGALSAVAATKSPIIFIGTGEHIDDFEPFKTQPFISKLLGMQFTLRDMYEQFQNIMKMGPFSQILGMIPGFGTDFMSKGNEQESMARLKKLMTIMDSMNDQELDSTDGAKVFSKQPGRIQRVARGSGVSTRDVQELLTQYTKFAQMVKKMXXXXXXXXXXXXXXXXXXXXXXX

[0446] SEQ ID NO: 268MXXADLGRKITSALRSLSNATIINEEVLNAMLKEVCTALLEADVNIKLVKQLRENVKSAIDLEEMASGLNKRKMIQHAVFKELVKLVDPGVKAWTPTKGKQNVIMFVGLQGSGKTTTCSKLAYYYQRKGWKTCLICADTFRAGAFDQLKQNATKARIPFYGSYHEQDPVIIASEGVEKFKNENFEIIIVDTSGRHHQHDSLFEEMLQVANAIQPDNIVYVMDASIGQACEAQAKAFK DKVDVASVIVTKLDGHAQGGGALSAVAATKSPIIFIGTGEHIDDFEPFKTQPFISKLLGMQ FTLRDMYEQFQNIMKMGPFSQILGMIPGFGTDFMSKGNEQESMARLKKLMTIMDSMND QELDSTDGAKVFSKQPGRIQRVARGSGVSTRDVQELLTQYTKFAQMVKKMXXXXXXXXXXXXXXXXXXXXXXX

[0447] SEQ ID NO: 269MXXADLGRKITSALRSLSNATIINEEVLNAMLKEVCTALLEADVNIKLVKQLRENVKSAIDLEEMASGLNKRKMIQHAVFKELVKLVDPGVKAWTPTKGKQNVIMFVGLQGSGKTTTCSKLAYYYQRKGWKTCLICADTFRAGAFDQLKQNATKARIPFYGSYEEEDPVIIASEGVEK FKNENFE111VDTSGRHEQEDSLFEEMLQVANA1QPDN1VYVMDAS1GQACEAQAKAFKD KVDVASVIVTKLDGHAEGGGALSAVAATKSPIIFIGTGEHIDDFEPFKTQPFISKLLGMQFT LRDMYEQFQNIMKMGPFSQILGMIPGFGTDFMSKGNEQESMARLKKLMTIMDSMNDQELDSTDGAKVFSKQPGRIQRVARGSGVSTRDVQELLTQYTKFAQMVKKMXXXXXXXXX XXXXXXXXXXXXXX

[0448] SEQ ID NO: 270MXXADLGRKITSALRSLSNATIINEEVLNAMLKEVCTALLEADVNIKLVKQLRENVKSAIDLEEMASGLNKRKMIQHAVFKELVKLVDPGVKAWTPTKGKQNVIMFVGLQGSGKTTTC SKLAYYYQRKGWKTCLICADTFRAGAFDQLKQNATKARIPFYGSYRERDPVIIASEGVE KFKNENFEIIIVDTSGRHRQRDSLFEEMLQVANAIQPDNIVYVMDASIGQACEAQAKAFK DKVDVASVIVTKLDGHARGGGALSAVAATKSPIIFIGTGEHIDDFEPFKTQPFISKLLGMQFTLRDMYEQFQNIMKMGPFSQILGMIPGFGTDFMSKGNEQESMARLKKLMTIMDSMNDQ ELDSTDGAKVFSKQPGRIQRVARGSGVSTRDVQELLTQYTKFAQMVKKMXXXXXXXX xxxxxxxxxxxxxxx

[0449] SEQ ID NO: 271MXXADLGRKITSALRSLSNATIINEEVLNAMLKEVCTALLEADVNIKLVKQLRENVKSAIDLEEMASGLNKRKMIQHAVFKELVKLVDPGVKAWTPTKGKQNVIMFVGLQGSGKTTTC SKLAYYYQRKGWKTCLICADTFRAGAFDQLKQNATKARIPFYGSYCECDPVIIASEGVE KFKNENFEIIIVDTSGRHCQCDSLFEEMLQVANAIQPDNIVYVMDASIGQACEAQAKAFK DKVDVASVIVTKLDGHACGGGALSAVAATKSPIIFIGTGEHIDDFEPFKTQPFISKLLGMQFTLRDMYEQFQNIMKMGPFSQILGMIPGFGTDFMSKGNEQESMARLKKLMTIMDSMNDQ ELDSTDGAKVFSKQPGRIQRVARGSGVSTRDVQELLTQYTKFAQMVKKMXXXXXXXXxxxxxxxxxxxxxxx

[0450] SEQ ID NO: 272MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKEHSVANRRLAFDRTRSKAAVGKLFHHLGHHYHHRPGGYLRILKCGFRAGDNAPMAYVELVHHHH

[0451] SEQ ID NO: 273MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKEHSVANRRLAFDRTRSKAAVGKLFHQLGHQYHQRPGGYLRILKCGFRAGDNAPMAYVELVHQHQ

[0452] SEQ ID NO: 274MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKEESVANRRLAFDRTRSKAAVGKLFEELGEEYEERPGGYLRILKCGFRAGDNAPMAYVELVEEE E

[0453] SEQ ID NO: 275MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKERSVANRRLAFDRTRSKAAVGKLFRRLGRRYRRRPGGYLRILKCGFRAGDNAPMAYVELVRRRR

[0454] SEQ ID NO: 276MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKEDSVANRRLAFDRTRSKAAVGKLFCCLGCCYCCRPGGYLRILKCGFRAGDNAPMAYVELVCCCC

[0455] SEQ ID NO: 277MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKEDSVANRRLAFDRTRSKAAVGKLFNDLGKRYANRPGGYLRILKCGFRAGDNAPMAYVELVEPEA

[0456] SEQ ID NO: 278MRHRKSGRHLSRTSAHRKAMFQNMAVSLFEHELIKTTLPKAKELRRVAEPLITLAKEDSVANRRLAFDRTRSKAAVGKLFCCLGPGYCCRPGGYLRILKCGFRAGDNAPMAYVELVDRPV

[0457] SEQ ID NO: 279MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNEHHHHHHLKVSTVFVNEGRSLKRIMPRAKGRADRIVKRSCHITVKVADK

[0458] SEQ ID NO: 280MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNEHQHQHQLKVSTVFVNEGRSLKRIMPRAKGRADRIVKRSCHITVKVADK

[0459] SEQ ID NO: 281MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNEGDDDDKLKVSTVFVNEGRSLKRIMPRAKGRADRIVKRSCHITVKVADK

[0460] SEQ ID NO: 282MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNEEEEEEELKVSTVFVNEGRSLKR1MPRAKGRADR1VKRSCH1TVKVADK

[0461] SEQ ID NO: 283MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNERRRRRRLKVSTVFVNEGRSLKRIMPRAKGRADRIVKRSCHITVKVADK

[0462] SEQ ID NO: 284MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNECCCCCCLKVSTVFVNEGRSLKRIMPRAKGRADRIVKRSCHITVKVADK

[0463] SEQ ID NO: 285MEVAAKLSGARISAQKARLVADQIRGKKVGEALNLLAFSSKKAAEIMKKVLESAVANAEHNECCPGCCLKVSTVFVNEGRSLKRIMPRAKGRADRIVKRSCHITVKVADK

[0464] SEQ ID NO: 286MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFHHHVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALHHHHHHHH

[0465] SEQ ID NO: 287MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFHQHVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALHQHQHQHQ

[0466] SEQ ID NO: 288MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALQPGDDDDK

[0467] SEQ ID NO: 289MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFEEEVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALEEEEEEEE

[0468] SEQ ID NO: 290MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFRRRVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALRRRRRRRR

[0469] SEQ ID NO: 291MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFCCCVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALCCCCCCCC

[0470] SEQ ID NO: 292MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALDYKDDDDK

[0471] SEQ ID NO: 293MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALQPGQEPEA

[0472] SEQ ID NO: 294MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALQPCCPGCC

[0473] SEQ ID NO: 295MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALDLYDDDDK

[0474] SEQ ID NO: 296MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALTQDPSRVG

[0475] SEQ ID NO: 297MNQERVFKVLLGPHISEKATGLADGKSQFVFKVATDATKLEIKKAVESLFSVKVQRVTTLNVKGKTKRTARGLGKRNDWKKAYIALWSHPQFEK

[0476] SEQ ID NO: 298MQKIRRDDEVIVIAGKDKHHHGKVLKVLADDRLVVHHVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNHHHHKADRVGFKVEDGKKIRVFKSTQHHHH

[0477] SEQ ID NO: 299MQKIRRDDEVIVIAGKDKHQHGKVLKVLADDRLVVHQVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNHQHQKADRVGFKVEDGKKIRVFKSTQHQHQ

[0478] SEQ ID NO: 300MQKIRRDDEVIVIAGKDKGKRGKVLKVLADDRLVVGGVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNDDDDKADRVGFKVEDGKKIRVFKSTQKPVQ

[0479] SEQ ID NO: 301MQKIRRDDEVIVIAGKDKEEEGKVLKVLADDRLVVEEVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNEEEEKADRVGFKVEDGKKIRVFKSTQEEEE

[0480] SEQ ID NO: 302MQKIRRDDEVIVIAGKDKEEEGKVLKVLADDRLVVEEVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNEEEEKADRVGFKVEDGKKIRVFKSTQEEEE

[0481] SEQ ID NO: 303MQK1RRDDEV1V1AGKDKRRRGKVLKVLADDRLVVRRVNL1KRHTKPNPMLGQQGG1VEKEAPLHVSNVAIFNRRRRKADRVGFKVEDGKKIRVFKSTQRRRR

[0482] SEQ ID NO: 304MQKIRRDDEVIVIAGKDKCCCGKVLKVLADDRLVVCCVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNCCCCKADRVGFKVEDGKKIRVFKSTQCCCC

[0483] SEQ ID NO: 305MQKIRRDDEVIVIAGKDKGKRGKVLKVLADDRLVVGGVNLIKRHTKPNPMLGQQGGIVEKEAPLHVSNVAIFNTETSKADRVGFKVEDGKKIRVFKSTQEPEA

[0484] SEQ ID NO: 306KANELREHHHGPVTGNNISHLLRDQFNLRMQKATGHLGHSHLLSQVKRDIARVKTVLNQQ

[0485] SEQ ID NO: 307KANELREHQHGPVTGNNISHLLRDQFNLRMQKATGHLGQSHLLSQVKRDIARVKTVLNQQ

[0486] SEQ ID NO: 308KANELREEEEGPVTGNNISHLLRDQFNLRMQKATGELGESHLLSQVKRDIARVKTVLNQQ

[0487] SEQ ID NO: 309KANELRERRRGPVTGNNISHLLRDQFNLRMQKATGRLGRSHLLSQVKRDIARVKTVLNQQ

[0488] SEQ ID NO: 310KANELRECCCGPVTGNNISHLLRDQFNLRMQKATGCLGCSHLLSQVKRDIARVKTVLNQQ

[0489] SEQ ID NO: 311MAVQQNKKSRSARDMRRSHDALESNALSVEKHHHHVHLRHHVSPDGFYRGHHHH

[0490] SEQ ID NO: 312MAVQQNKKSRSARDMRRSHDALESNALSVEKHQHQVHLRHHVSPDGFYRGHQHQ

[0491] SEQ ID NO: 313MAVQQNKKSRSARDMRRSHDALESNALSVEKEEEEVHLRHHVSPDGFYRGEEEE

[0492] SEQ ID NO: 314MAVQQNKKSRSARDMRRSHDALESNALSVEKRRRRVHLRHHVSPDGFYRGRRRR

[0493] SEQ ID NO: 315MAVQQNKKSRSARDMRRSHDALESNALSVEKCCCCVHLRHHVSPDGFYRGCCCC

[0494] SEQ ID NO: 316MAVQQNKKSRSARDMRRSHDALESNALSVEKSTGEVHLRHHVSPDGFYRGEPEA

[0495] SEQ ID NO: 317MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDHHHHHHHHHHHHHHHHHHHTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKPEVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIKLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKHHHHHNTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITY VSVESKQ

[0496] SEQ ID NO: 318MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDHQHQHQHQHQHQHQHQHQHTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKPEVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIKLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKHQHQHNTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0497] SEQ ID NO: 319MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDDDDDKDDDDKDDDDKK PKMTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKK PEVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWG IKLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKDDDDKNTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITY VSVESKQ

[0498] SEQ ID NO: 320MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDEEEEEEEEEEEEEEEEEEETFYKLLERR1ATLPKNYTQEMFEQKLGQLEDM1KEGE1VDVRA1VATQPWV1KLKKKPEVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIKL RAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKEEEEE NTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVE SKQ

[0499] SEQ ID NO: 321MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDRRRRRRRRRRRRRRRRRRRTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKP EVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGI KLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLK LDENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0500] SEQ ID NO: 322MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDCCCCCCCCCCCCCCCCCCCTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKP EVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGI KLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLK LDENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0501] SEQ ID NO: 323MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPG LTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDYKDDDDKDYKDDDDKK PKMTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKK PEVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIKLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLK LDENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYV SVESKQ

[0502] SEQ ID NO: 324MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKE1FMPVT11ETPPLRVFG1RAYRMGYLGLETATEV1VPDCCPGCCCCPGCCCCPGCCMTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKPE VMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIK LRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLKL DENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0503] SEQ ID NO: 325MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDLYDDDDKDLYDDDDKKPKMTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKP EVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGI KLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLK LDENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0504] SEQ ID NO: 326MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDTQDPSRVGTQDPSRVGPKMTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKPE VMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIK LRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLKL DENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0505] SEQ ID NO: 327MGKVHRPRRGSLGFSPRKRAKSIVPRIRSWPKETEVRMLGFAGYKAGMTHILMIDDEPGLTNGKEIFMPVTIIETPPLRVFGIRAYRMGYLGLETATEVIVPDWSHPQFEKWSHPQFEKPKMTFYKLLERRIATLPKNYTQEMFEQKLGQLEDMIKEGEIVDVRAIVATQPWVIKLKKKPEVMEYAIGGTSVEEKFNYIKEKLGKELRVGEVLKEGELLDVIAVTKGKGTQGPVKRWGIKLRAHKDSKGRRKVGSIGPWHPARVMWTVPMAGQMGFHHRTELNKRLIAIGENGKLKLDENTEIEITPKGGFPHYGIVRSDFMMIAGSVPGAIKRIIRVRPAIRPPKKKPPVQRPQITYVSVESKQ

[0506] SEQ ID NO: 328MKISSKQPRKQRKFLYNAPLHVRQKLMSAPLSRELREKYKVRNLPVRVHDHVR1MRGDYKGHEGKVVEVDLKRYRIYVHHATLRKTHHHHVFYPIHPSNVMIIELHHHDEKRKKIIERRAG

[0507] SEQ ID NO: 329MKISSKQPRKQRKFLYNAPLHVRQKLMSAPLSRELREKYKVRNLPVRVHDQVRIMRGDYKGHEGKVVEVDLKRYRIYVHQATLRKTHQHQVFYPIHPSNVMIIELHQHDEKRKKIIERRAG

[0508] SEQ ID NO: 330MKISSKQPRKQRKFLYNAPLHVRQKLMSAPLSRELREKYKVRNLPVRVEDEVRIMRGDYKGHEGKVVEVDLKRYRIYVEEATLRKTEEEEVFYPIHPSNVMIIELEEEDEKRKKIIERRAG

[0509] SEQ ID NO: 331MKISSKQPRKQRKFLYNAPLHVRQKLMSAPLSRELREKYKVRNLPVRVRDRVRIMRGDYKGHEGKVVEVDLKRYRIYVRRATLRKTRRRRVFYPIHPSNVMIIELRREDEKRKKIIERRAG

[0510] SEQ ID NO: 332MKISSKQPRKQRKFLYNAPLHVRQKLMSAPLSRELREKYKVRNLPVRVCDCVRIMRGDYKGHEGKVVEVDLKRYRIYVCCATLRKTCCCCVFYPIHPSNVMIIELCCEDEKRKKIIERRAG

[0511] SEQ ID NO: 333MKPSEIREMSIEEIDAKIRELRLQLAKERGLLTMGHHHHHPMVIRNLRRDIARLLTIKKEKLREREKGKVKKIll

[0512] SEQ ID NO: 334MKPSEIREMSIEEIDAKIRELRLQLAKERGLLTMGHQHQHPMVIRNLRRDIARLLTIKKEKLREREKGKVKK

[0513] SEQ ID NO: 335MKPSEIREMSIEEIDAKIRELRLQLAKERGLLTMGDDDDKPMVIRNLRRDIARLLTIKKEKLREREKGKVKK

[0514] SEQ ID NO: 336MKPSEIREMSIEEIDAKIRELRLQLAKERGLLTMGEEEEEPMVIRNLRRDIARLLTIKKEKLREREKGKVKK

[0515] SEQ ID NO: 337MKPSEIREMSIEEIDAKIRELRLQLAKERGLLTMGRRRRRPMVIRNLRRDIARLLTIKKEKLREREKGKVKK

[0516] SEQ ID NO: 338MKPSEIREMSIEEIDAKIRELRLQLAKERGLLTMGCCCCCPMVIRNLRRDIARLLTIKKEKLREREKGKVKK

[0517] SEQ ID NO: 339MAIHHHHHVIFTVPIKKIKKIVPRWKRAPRAVKFVREFVARHAKAQEVIIDPKVNEKIWERGIEKPPSKLRVKVKVEEHHHHHEHTVRIAYVTLA

[0518] SEQ ID NO: 340MAIHQHQHVIFTVPIKKIKKIVPRWKRAPRAVKFVREFVARHAKAQEVIIDPKVNEKIWERGIEKPPSKLRVKVKVEEHQHQHEQTVRIAYVTLA

[0519] SEQ ID NO: 341MAIDDDDKVIFTVPIKKIKKIVPRWKRAPRAVKFVREFVARHAKAQEVIIDPKVNEKIWERGIEKPPSKLRVKVKVEEDDDDKEETVRIAYVTLA

[0520] SEQ ID NO: 342MAIEEEEEVIFTVPIKKIKKIVPRWKRAPRAVKFVREFVARHAKAQEVIIDPKVNEKIWERGIEKPPSKLRVKVKVEEEEEEEEETVRIAYVTLA

[0521] SEQ ID NO: 343MAIRRRRRVIFTVPIKKIKKIVPRWKRAPRAVKFVREFVARHAKAQEVIIDPKVNEKIWERGIEKPPSKLRVKVKVEERRRRRERTVRIAYVTLA

[0522] SEQ ID NO: 344MAICCCCCVIFTVPIKKIKKIVPRWKRAPRAVKFVREFVARHAKAQEVIIDPKVNEKIWERGIEKPPSKLRVKVKVEECCCCCECTVRIAYVTLA

[0523] SEQ ID NO: 345MARYGATSTNHHHSASARGSYLRVSFKNTRETAQAINGWELTKAQKYLEQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAAANAEAKGHHHHHLYVSHIQVNQAPKQRRRTYRAHGRINKYESSPSHIELVVTEKEEAVAKAAEKKVVRLTS

[0524] SEQ ID NO: 346MARYGATSTNHQHSASARGSYLRVSFKNTRETAQAINGWELTKAQKYLEQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAAANAEAKGHQHQHLYVSH1QVNQAPKQRRRTYRAHGR1NKYESSPSH1ELVVTEKEEAVAKAAEKKVVRLTS

[0525] SEQ ID NO: 347MARYGATSTNPAKSASARGSYLRVSFKNTRETAQAINGWELTKAQKYLEQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAAANAEAKGDDDDKLYVSHIQVNQAPKQRRRTYRAHGRINKYESSPSHIELVVTEKEEAVAKAAEKKVVRLTS

[0526] SEQ ID NO: 348MARYGATSTNEEESASARGSYLRVSFKNTRETAQAINGWELTKAQKYLEQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAAANAEAKGEEEEELYVSHIQVNQAPKQRRRTYRAHGRINKYESSPSHIELVVTEKEEAVAKAAEKKVVRLTS

[0527] SEQ ID NO: 349MARYGATSTNRRRSASARGSYLRVSFKNTRETAQAINGWELTKAQKYLEQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAAANAEAKGRRRRRLYVSHIQVNQAPKQRRRTYRAHGRINKYESSPSHIELVVTEKEEAVAKAAEKKVVRLTS

[0528] SEQ ID NO: 350MARYGATSTNCCCSASARGSYLRVSFKNTRETAQAINGWELTKAQKYLEQVLDHQRAIPFRRFNSSIGRTAQGKEFGVTKARWPAKSVKFVQGLLQNAAANAEAKGCCCCCLYVSHIQVNQAPKQRRRTYRAHGRINKYESSPSHIELVVTEKEEAVAKAAEKKVVRLTS

[0529] SEQ ID NO: 351MAPNTSRKQKIAKTFTHDHSSPTENGVHHPASYAKYLIDHIKVEGAVGNLGNAVTVTEHHHVVTVVSTAKFSGKYLKYLTKKYLKKNQLRDWIRFVSTKTNEYRLAFYQVTPEEDEEEDEE

[0530] SEQ ID NO: 352MAPNTSRKQKIAKTFTHDQSSPTENGVHQPASYAKYLIDHIKVEGAVGNLGNAVTVTEHQHVVTVVSTAKFSGKYLKYLTKKYLKKNQLRDWIRFVSTKTNEYRLAFYQVTPEEDEEEDEE

[0531] SEQ ID NO: 353MAPNTSRKQKIAKTFTEDESSPTENGVEEPASYAKYLIDHIKVEGAVGNLGNAVTVTEEEEVVTVVSTAKFSGKYLKYLTKKYLKKNQLRDWIRFVSTKTNEYRLAFYQVTPEEDEEEDEE

[0532] SEQ ID NO: 354MAPNTSRKQKIAKTFTRDRSSPTENGVRRPASYAKYLIDHIKVEGAVGNLGNAVTVTERRRVVTVVSTAKFSGKYLKYLTKKYLKKNQLRDW1RFVSTKTNEYRLAFYQVTPEEDEEEDEE

[0533] SEQ ID NO: 355MAPNTSRKQKIAKTFTCDCSSPTENGVCCPASYAKYLIDHIKVEGAVGNLGNAVTVTECCCVVTVVSTAKFSGKYLKYLTKKYLKKNQLRDWIRFVSTKTNEYRLAFYQVTPEEDEEEDEE

[0534] SEQ ID NO: 356MAKQSLDVSSDRRKARKAYFTAPSSQRRVLLSAPLSKELRAQYGIKALPIRRDDEVLVVRGSKKHHHGKISSVYRLKFAVQVHHVTKEKVNGASVPINLHPHHLVITHLHLDKDRKALIQRKG

[0535] SEQ ID NO: 357MAKQSLDVSSDRRKARKAYFTAPSSQRRVLLSAPLSKELRAQYGIKALPIRRDDEVLVVRGSKKHQHGKISSVYRLKFAVQVHQVTKEKVNGASVPINLHPHQLVITHLHLDKDRKALIQRKG

[0536] SEQ ID NO: 358MAKQSLDVSSDRRKARKAYFTAPSSQRRVLLSAPLSKELRAQYGIKALPIRRDDEVLVVRGSKKEEEGKISSVYRLKFAVQVEEVTKEKVNGASVPINLHPEELVITELHLDKDRKALIQR KG

[0537] SEQ ID NO: 359MAKQSLDVSSDRRKARKAYFTAPSSQRRVLLSAPLSKELRAQYGIKALPIRRDDEVLVVRGSKKRRRGKISSVYRLKFAVQVRRVTKEKVNGASVPINLHPRRLVITRLHLDKDRKALIQ RKG

[0538] SEQ ID NO: 360MAKQSLDVSSDRRKARKAYFTAPSSQRRVLLSAPLSKELRAQYGIKALPIRRDDEVLVVRGSKKCCCGKISSVYRLKFAVQVCCVTKEKVNGASVPINLHPCCLVITCLHLDKDRKALIQ RKG

[0539] SEQ ID NO: 361MAGHHHHVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKHHEEEDAKNPLFSYVEPVLVASAKGLQTHHHHHHH

[0540] SEQ ID NO: 362MAGHQHQVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQA1WKRGVKGVEYRLRLR1SRKHQEEEDAKNPLFSYVEPVLVASAKGLQTHQHQHQH

[0541] SEQ ID NO: 363MAGLKDVVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKRNEEEDAKNPLFSYVEPVLVASAKGLQTVVDDDDK

[0542] SEQ ID NO: 364MAGEEEEVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKEEEEEDAKNPLFSYVEPVLVASAKGLQTEEEEEEE

[0543] SEQ ID NO: 365MAGRRRRVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKRREEEDAKNPLFSYVEPVLVASAKGLQTRRRRRRR

[0544] SEQ ID NO: 366MAGCCCCVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKCCEEEDAKNPLFSYVEPVLVASAKGLQTCCCCCCC

[0545] SEQ ID NO: 367MAGLKDVVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKRNEEEDAKNPLFSYVEPVLVASAKGLQTEPEAEDA

[0546] SEQ ID NO: 368MAGLKDVVTREYTINLHKRLHGVSFKKRAPRAVKEIKKFAKLHMGTDDVRLAPELNQAIWKRGVKGVEYRLRLRISRKRNEEEDAKNPLFSYVEPVLVASAKGLQTQDPSRVG

[0547] SEQ ID NO: 369MAREITDIKQFLELTRRADVKTATVKINKKHHHHHHHHHQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKHHHH

[0548] SEQ ID NO: 370MAREITDIKQFLELTRRADVKTATVKINKKHQHQHQHQHQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKHQHQ

[0549] SEQ ID NO: 371MAREITDIKQFLELTRRADVKTATVKINKKLDDDDKPFRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKVNRL

[0550] SEQ ID NO: 372MAREITDIKQFLELTRRADVKTATVKINKKEEEEEEEEEQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKEEEE

[0551] SEQ ID NO: 373MAREITDIKQFLELTRRADVKTATVKINKKRRRRRRRRRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKRRRR

[0552] SEQ ID NO: 374MAREITDIKQFLELTRRADVKTATVKINKKCCCCCCCCCQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKCCCC

[0553] SEQ ID NO: 375MAREITDIKQFLELTRRADVKTATVKINKKDYKDDDDKRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKVNRL

[0554] SEQ ID NO: 376MAREITDIKQFLELTRRADVKTATVKINKKLNKAGKPFRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKEPEA

[0555] SEQ ID NO: 377MAREITDIKQFLELTRRADVKTATVKINKKLCCPGCCFRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKVNRL

[0556] SEQ ID NO: 378MAREITDIKQFLELTRRADVKTATVKINKKDLYDDDDKRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKVNRL

[0557] SEQ ID NO: 379MAREITDIKQFLELTRRADVKTATVKINKKTQDPSRVGRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKVNRL

[0558] SEQ ID NO: 380MAREITDIKQFLELTRRADVKTATVKINKKWSHPQFEKRQTKFKVRGSSSLYTLVINDAGKAKKLIQSLPPTLKVNRL

[0559] SEQ ID NO: 381MGFFKKKEKKAEFDVFRLYGLHHPEPGKCDEITTRGYSISMSVDKARRVIDQIRGRSYAETLMILELMPYRACYPIFKLIYSAAANASHNKHFHHHNLIISKAEVNKGITLKKVKPRARGRSYMIKRPTCHITIVLRDITHFDSYDKFLESLTPKKLIALLGLMSTGRRRELLCGRFRENHKIKSFLYKIALFKRYEVM

[0560] SEQ ID NO: 382MGFFKKKEKKAEFDVFRLYGLHHPEPGKCDEITTRGYSISMSVDKARRVIDQIRGRSYAETLM1LELMPYRACYP1FKL1YSAAANASHNKHFQHQNL11SKAEVNKG1TLKKVKPRARGRSYMIKRPTCHITIVLRDITHFDSYDKFLESLTPKKLIALLGLMSTGRRRELLCGRFRENHKIKSFLYKIALFKRYEVM

[0561] SEQ ID NO: 383MGFFKKKEKKAEFDVFRLYGLHHPEPGKCDEITTRGYSISMSVDKARRVIDQIRGRSYAETLMILELMPYRACYPIFKLIYSAAANASHNKEFEEENLIISKAEVNKGITLKKVKPRARGRSYMIKRPTCHITIVLRDITHFDSYDKFLESLTPKKLIALLGLMSTGRRRELLCGRFRENHKIKSFLYKIALFKRYEVM

[0562] SEQ ID NO: 384MGFFKKKEKKAEFDVFRLYGLHHPEPGKCDEITTRGYSISMSVDKARRVIDQIRGRSYAETLMILELMPYRACYPIFKLIYSAAANASHNKRFRRRNLIISKAEVNKGITLKKVKPRARGRSYMIKRPTCHITIVLRDITHFDSYDKFLESLTPKKLIALLGLMSTGRRRELLCGRFRENHKIKSFLYKIALFKRYEVM

[0563] SEQ ID NO: 385MGFFKKKEKKAEFDVFRLYGLHHPEPGKCDEITTRGYSISMSVDKARRVIDQIRGRSYAETLMILELMPYRACYPIFKLIYSAAANASHNKCFCCCNLIISKAEVNKGITLKKVKPRARGRSYMIKRPTCHITIVLRDITHFDSYDKFLESLTPKKLIALLGLMSTGRRRELLCGRFRENHKIKSFLYKIALFKRYEVM

[0564] SEQ ID NO: 386ATEAVAPTTEEAATSQPKTSKKAKKLKYPRRILDVYQILQSPIITEAAIKNIADENSLLFTVDVRADKKMIREAISNFFGVKVRKVNTLIRPDGTKKAYIMLNHHHHASELAKHHHHFPG GN

[0565] SEQ ID NO: 387ATEAVAPTTEEAATSQPKTSKKAKKLKYPRRILDVYQILQSPIITEAAIKNIADENSLLFTVDVRADKKMIREAISNFFGVKVRKVNTLIRPDGTKKAYIMLNHQHQASELAKHQHQFPG GN

[0566] SEQ ID NO: 388ATEAVAPTTEEAATSQPKTSKKAKKLKYPRRILDVYQILQSPIITEAAIKNIADENSLLFTVDVRADKKMIREAISNFFGVKVRKVNTLIRPDGTKKAYIMLNEEEEASELAKEEEEFPGGN

[0567] SEQ ID NO: 389ATEAVAPTTEEAATSQPKTSKKAKKLKYPRRILDVYQILQSPIITEAAIKNIADENSLLFTVDVRADKKM1REA1SNFFGVKVRKVNTL1RPDGTKKAY1MLNRRRRASELAKRRRRFPGGN

[0568] SEQ ID NO: 390ATEAVAPTTEEAATSQPKTSKKAKKLKYPRRILDVYQILQSPIITEAAIKNIADENSLLFTVDVRADKKMIREAISNFFGVKVRKVNTLIRPDGTKKAYIMLNCCCCASELAKCCCCFPGGN

[0569] SEQ ID NO: 391ATEAVAPTTEEAATSQPKTSKKAKKLKYPRRILDVYQILQSPIITEAAIKNIADENSLLFTVDVRADKKMIREAISNFFGVKVRKVNTLIRPDGTKKAYIMLNKEYNASELAKEPEAFPGGN

[0570] SEQ ID NO: 392RIKRWERKDCKPNSLPKLHKRHVKVGDTVKVISGGEKGKIGEISKIHKHNSTVIIKDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDHHHHWKEIHHKKESETAVAVAA

[0571] SEQ ID NO: 393RIKRWERKDCKPNSLPKLHKRHVKVGDTVKVISGGEKGKIGEISKIHKHNSTVIIKDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDHQHQWKEIHQKKESETAVAVAA

[0572] SEQ ID NO: 394RIKRWERKDCKPNSLPKLHKRHVKVGDTVKVISGGEKGKIGEISKIHKHNSTVIIKDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDDDDKWKEIQNKKESETAVAVAA

[0573] SEQ ID NO: 395RIKRWERKDCKPNSLPKLHKRHVKVGDTVKVISGGEKGKIGEISKIHKHNSTVIIKDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDEEEEWKEIEEKKESETAVAVAA

[0574] SEQ ID NO: 396RIKRWERKDCKPNSLPKLHKRHVKVGDTVKVISGGEKGKIGEISKIHKHNSTVIIKDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDRRRRWKEIRRKKESETAVAVAA

[0575] SEQ ID NO: 397R1KRWERKDCKPNSLPKLHKRHVKVGDTVKV1SGGEKGK1GE1SK1HKHNSTV11KDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDCCCCWKEICCKKESETAVAVAA

[0576] SEQ ID NO: 398RIKRWERKDCKPNSLPKLHKRHVKVGDTVKVISGGEKGKIGEISKIHKHNSTVIIKDLNFKTKHVKSKEEGEQGQIIKIEAAIHSSNVMLILKEQEVADRVGHKILEDVRKVRYLIKTGEIVDEPEAWKEIQNKKESETAVAVAA

[0577] SEQ ID NO: 399KKEDELKELRTKTNEQLNEEILQLKGELFMLRLQRSARENFKPSDFGRMRKRVARMLTVKREREIEQGVGKRLSRKLDKAWKRSIVVRPHHHHKKLQEKATAEAEAEKA

[0578] SEQ ID NO: 400KKEDELKELRTKTNEQLNEEILQLKGELFMLRLQRSARENFKPSDFGRMRKRVARMLTVKREREIEQGVGKRLSRKLDKAWKRSIVVRPHQHQKKLQEKATAEAEAEKA

[0579] SEQ ID NO: 401KKEDELKELRTKTNEQLNEEILQLKGELFMLRLQRSARENFKPSDFGRMRKRVARMLTVKREREIEQGVGKRLSRKLDKAWKRSIVVRPEEEEKKLQEKATAEAEAEKA

[0580] SEQ ID NO: 402KKEDELKELRTKTNEQLNEEILQLKGELFMLRLQRSARENFKPSDFGRMRKRVARMLTVKREREIEQGVGKRLSRKLDKAWKRSIVVRPRRRRKKLQEKATAEAEAEKA

[0581] SEQ ID NO:403KKEDELKELRTKTNEQLNEEILQLKGELFMLRLQRSARENFKPSDFGRMRKRVARMLTVKREREIEQGVGKRLSRKLDKAWKRSIVVRPCCCCKKLQEKATAEAEAEKA

[0582] SEQ ID NO: 404KKEDELKELRTKTNEQLNEEILQLKGELFMLRLQRSARENFKPSDFGRMRKRVARMLTVKREREIEQGVGKRLSRKLDKAWKRSIVVRPEPEAKKLQEKATAEAEAEKA

[0583] SEQ ID NO: 405MGYRKLGRTSDQRKAMLRDLATSLIISERIETTEARAKEVRSVVEKLITLGKKGDLASRRNAAKTLRNVEILNHHHTTQTALQKLFGEIAERYHHHQGGYTRILKQGPRRGDGAESVIIELV

[0584] SEQ ID NO: 406MGYRKLGRTSDQRKAMLRDLATSLIISERIETTEARAKEVRSVVEKLITLGKKGDLASRRNAAKTLRNVE1LNHQHTTQTALQKLFGE1AERYHQHQGGYTR1LKQGPRRGDGAESV11ELV

[0585] SEQ ID NO: 407MGYRKLGRTSDQRKAMLRDLATSLIISERIETTEARAKEVRSVVEKLITLGKKGDLASRRNAAKTLRNVEILNEEETTQTALQKLFGEIAERYEEEQGGYTRILKQGPRRGDGAESVIIELV

[0586] SEQ ID NO: 408MGYRKLGRTSDQRKAMLRDLATSLIISERIETTEARAKEVRSVVEKLITLGKKGDLASRRNAAKTLRNVEILNRRRTTQTALQKLFGEIAERYRRRQGGYTRILKQGPRRGDGAESVIIELV

[0587] SEQ ID NO: 409MGYRKLGRTSDQRKAMLRDLATSLIISERIETTEARAKEVRSVVEKLITLGKKGDLASRRNAAKTLRNVEILNCCCTTQTALQKLFGEIAERYCCCQGGYTRILKQGPRRGDGAESVIIELV

[0588] SEQ ID NO: 410MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYHMHHHHLVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSHHHHHHHHH

[0589] SEQ ID NO: 411MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYHMQHQHLVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSHQHQHQHQH

[0590] SEQ ID NO: 412MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMDDDKLVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSDDDDKAKEA

[0591] SEQ ID NO: 413MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYEMEEEELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSEEEEEEEEE

[0592] SEQ ID NO: 414MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYRMRRRELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSRRRRRRRRR

[0593] SEQ ID NO: 415MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYCMCCCCLVVKEAYANEGPTLKRFRPRAQGRASA1NKRTSH1T1VVSCCCCCCCCC

[0594] SEQ ID NO: 416MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMNTDELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSDYKDDDDKA

[0595] SEQ ID NO: 417MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMNTDELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSDGKEEEPEA

[0596] SEQ ID NO: 418MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMNTDELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSCCPGCCKEA

[0597] SEQ ID NO: 419MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMNTDELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSDLYDDDDKA

[0598] SEQ ID NO: 420MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMNTDELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSTQDPSRVGA

[0599] SEQ ID NO: 421MEAKAVARTIRIAPRKVRLVLDLIRGKNAAEAIAILKLTNKASSPVIEKVLMSALANAEHNYDMNTDELVVKEAYANEGPTLKRFRPRAQGRASAINKRTSHITIVVSWSHPQFEKA

[0600] SEQ ID NO: 422MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFHVHVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSHHHHH

[0601] SEQ ID NO: 423MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFHVQVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSHQHQH

[0602] SEQ ID NO: 424MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFNVKVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSDDDDK

[0603] SEQ ID NO: 425MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFEVEVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSEEEEE

[0604] SEQ ID NO: 426MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFRVRVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSRRRRR

[0605] SEQ ID NO: 427MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFCVCVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSCCCCC

[0606] SEQ ID NO: 428MEARDILKRPVITEKSSEAMAEDKYTFDVDTRVNKTQVKMAVEEIFNVKVASVNIMNYKPKKKRMGRYQGYTNKRRKAIVTLKEGSIEPEA

[0607] SEQ ID NO: 429MHIKKHDHVKVIAGKDKHKHHHVIATLPKKDRVVVHHVNIMKKHQKPTQHHHHHHHHHTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIHHHH

[0608] SEQ ID NO: 430MHIKKGDNVKVIAGKDKHKQHQVIATLPKKDRVVVHQVNIMKKHQKPTQHQHQHQHQHTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIHQHQ

[0609] SEQ ID NO: 431MHIKKGDNVKVIAGKDKGKEGKVIATLPKKDRVVVEGVNIMKKHQKPTQDDDDKGILETEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIKSNN

[0610] SEQ ID NO: 432MHIKKGDNVKVIAGKDKEKEGKVIATLPKKDRVVVEEVNIMKKHQKPTQEEEEEEEEETEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIEEEE

[0611] SEQ ID NO: 433MHIKKRDRVKVIAGKDKRKRRRVIATLPKKDRVVVRRVNIMKKHQKPTQRRRRRRRRRTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIRRRR

[0612] SEQ ID NO: 434MHIKKCDCVKVIAGKDKCKCCCVIATLPKKDRVVVCCVNIMKKHQKPTQCCCCCCCCCTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEICCCC

[0613] SEQ ID NO: 435MHIKKGDNVKVIAGKDKGKEGKVIATLPKKDRVVVEGVNIMKKHQKPTQLDYKDDDDKTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIKSNN

[0614] SEQ ID NO: 436MH1KKGDNVKV1AGKDKGKEGKV1ATLPKKDRVVVEGVN1MKKHQKPTQLNPEGG1LETEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIEPEA

[0615] SEQ ID NO: 437MHIKKGDNVKVIAGKDKGKEGKVIATLPKKDRVVVEGVNIMKKHQKPTQLCCPGCCLETEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIKSNN

[0616] SEQ ID NO: 438MHIKKGDNVKVIAGKDKGKEGKVIATLPKKDRVVVEGVNIMKKHQKPTQLDLYDDDDKTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIKSNN

[0617] SEQ ID NO: 439MHIKKGDNVKVIAGKDKGKEGKVIATLPKKDRVVVEGVNIMKKHQKPTQLTQDPSRVGTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIKSNN

[0618] SEQ ID NO: 440MHIKKGDNVKVIAGKDKGKEGKVIATLPKKDRVVVEGVNIMKKHQKPTQLWSHPQFEKTEAAIHVSNVQLLDPKTNEPTRVGYKFVDGKKVRIAKKSGEEIKSNN

[0619] SEQ ID NO: 441MHHHHKAHHHRHLTTSEIEEQIKSSKEELFNLRFQLATGQLEETARIRTVRKTIARLKTVAREREIEQSKANQ

[0620] SEQ ID NO: 442MHQHQKAHQHRQLTTSEIEEQIKSSKEELFNLRFQLATGQLEETARIRTVRKTIARLKTVAREREIEQSKANQ

[0621] SEQ ID NO: 443MDDDDKAKEIRDLTTSEIEEQIKSSKEELFNLRFQLATGQLEETARIRTVRKTIARLKTVAREREIEQSKANQ

[0622] SEQ ID NO: 444MEEEEKAEEERELTTSEIEEQIKSSKEELFNLRFQLATGQLEETARIRTVRKTIARLKTVAREREIEQSKANQ

[0623] SEQ ID NO: 445MRRRRKARRRRRLTTSEIEEQIKSSKEELFNLRFQLATGQLEETARIRTVRKTIARLKTVAREREIEQSKANQ

[0624] SEQ ID NO: 446MCCCCKACCCRCLTTSEIEEQIKSSKEELFNLRFQLATGQLEETARIRTVRKTIARLKTVAREREIEQSKANQ

[0625] SEQ ID NO: 447MAPVKKLVAKGGKKKKQVLKFTLDCTHPVHHHHMHAHNFEQFLQERIKVNGKAGNLGGGVVTIERHHSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFQINQDEEEEEDED

[0626] SEQ ID NO: 448MAPVKKLVAKGGKKKKQVLKFTLDCTHPVHQHQMHAQNFEQFLQERIKVNGKAGNLGGGVVTIERHQSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFQINQDEEEEEDED

[0627] SEQ ID NO: 449MAPVKKLVAKGGKKKKQVLKFTLDCTHPVEEEEMEAENFEQFLQERIKVNGKAGNLGGGVVTIEREESKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFQINQDEEEEEDED

[0628] SEQ ID NO: 450MAPVKKLVAKGGKKKKQVLKFTLDCTHPVRRRRMRARNFEQFLQERIKVNGKAGNLGGGVVTIERRRSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFQINQDEEEEEDED

[0629] SEQ ID NO: 451MAPVKKLVAKGGKKKKQVLKFTLDCTHPVCCCCMCACNFEQFLQERIKVNGKAGNLGGGVVTIERCCSKITVTSEVPFSKRYLKYLTKKYLKKNNLRDWLRVVANSKESYELRYFQI NQDEEEEEDED

[0630] SEQ ID NO: 452MVRYSLDPHHHHHSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKHLHVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILHHHH

[0631] SEQ ID NO: 453MVRYSLDPHQHQHSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFHQHQLKNAESNAELKGLDVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKE

[0632] SEQ ID NO: 454MVRYSLDPDDDDKSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKGLDVDSLV1EHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKE

[0633] SEQ ID NO: 455MVRYSLDPEEEEESCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFEEEELKNAESNAELKGLDVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKE

[0634] SEQ ID NO: 456MVRYSLDPRRRRRSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFRRRRLKNAESNAELKGLDVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKE

[0635] SEQ ID NO: 457MVRYSLDPCCCCCSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFCCCCLKNAESNAELKGLDVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILTEKE

[0636] SEQ ID NO: 458MVRYSLDPENPTKSCKSRGSNLRVHFKNTRETAQAIKGMHIRKATKYLKDVTLQKQCVPFRRYNGGVGRCAQAKQWGWTQGRWPKKSAEFLLHMLKNAESNAELKGLDVDSLVIEHIQVNKAPKMRRRTYRAHGRINPYMSSPCHIEMILEPEA

[0637] SEQ ID NO: 459MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYDIDVAKVNTLIRPDGEKKAYVRLAHHYHALDVANKHHHH

[0638] SEQ ID NO: 460MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYDIDVAKVNTLIRPDGEKKAYVRLAHQYHALDVANKHQHQ

[0639] SEQ ID NO: 461MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYDIDVAKVNTLIRPDGEKKAYVRLAEEYEALDVANKEEEE

[0640] SEQ ID NO: 462MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKK1RTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYDIDVAKVNTLIRPDGEKKAYVRLARRYRALDVANKRRRR

[0641] SEQ ID NO: 463MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYDIDVAKVNTLIRPDGEKKAYVRLACCYCALDVANKCCCC

[0642] SEQ ID NO: 464MAPKAKKEAPAPPKAEAKAKALKAKKAVLKGVHSHKKKKIRTSPTFRRPKTLRLRRQPKYPRKSAPRRNKLDHYAIIKFPLTTESAMKKIEDNNTLVFIVDVKANKHQIKQAVKKLYDIDVAKVNTLIRPDGEKKAYVRLAPDYDALDVANKEPEA

[0643] SEQ ID NO: 465MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKHDHVQVVRGHYKGHHHGHVHQVYRKKYVIYIERVQREKANHTHVHVGIHPSKVVITHLHHDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0644] SEQ ID NO: 466MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKHDQVQVVRGHYKGHQHGQVHQVYRKKYVIYIERVQREKANHTQVHVGIHPSKVVITHLQHDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0645] SEQ ID NO: 467MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKEDEVQVVRGHYKGEEEGEVEQVYRKKYVIYIERVQREKANETEVHVGIHPSKVVITELEEDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0646] SEQ ID NO: 468MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKRDRVQVVRGHYKGRRRGRVRQVYRKKYVIYIERVQREKANRTRVHVGIHPSKVVITRLRRDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0647] SEQ ID NO: 469MKFNPFVTSDRSKNRKRHFNAPSHIRRKIMSSPLSKELRQKYNVRSMPIRKCDEVQVVRGHYKGCCCGCVCQVYRKKYVIYIERVQREKANCTCVHVGIHPSKVVITCLCCDKDRKKILERKAKSRQVGKEKGKYKEETIEKMQE

[0648] SEQ ID NO: 470MKTAYDVILAPVLSEKAYAGFHEGKYTFWVHPKATKTEIKNAVETAFKVKVVKVNTLHVRGKKKRLGRYLGKRPDRKKAIVQVAHHQKIEALEGLI

[0649] SEQ ID NO: 471MKTAYDVILAPVLSEKAYAGFHEGKYTFWVHPKATKTEIKNAVETAFKVKVVKVNTLHVRGKKKRLGRYLGKRPDRKKAIVQVAQHQKIEALEGLI

[0650] SEQ ID NO: 472MKTAYDVILAPVLSEKAYAGFEEGKYTFWVHPKATKTEIKNAVETAFKVKVVKVNTLHVRGKKKRLGRYLGKRPDRKKAIVQVAEEQKIEALEGLI

[0651] SEQ ID NO: 473MKTAYDVILAPVLSEKAYAGFREGKYTFWVHPKATKTEIKNAVETAFKVKVVKVNTLHVRGKKKRLGRYLGKRPDRKKAIVQVARRQKIEALEGLI

[0652] SEQ ID NO: 474MRVKMHVKKGDTVLVASGKYKGRVGKVKEVLPKKYAVIVEGVNIVKKAVRVSPHHHHHGFIEKEAPLHASKVRPICPACGKPTRVRKKFLENGKKIRVCAKCGGALDTEE

[0653] SEQ ID NO: 475MRVKMHVKKGDTVLVASGKYKGRVGKVKEVLPKKYAVIVEGVNIVKKAVRVSPHQHQHGFIEKEAPLHASKVRPICPACGKPTRVRKKFLENGKKIRVCAKCGGALDTEE

[0654] SEQ ID NO: 476MRVKMHVKKGDTVLVASGKYKGRVGKVKEVLPKKYAVIVEGVNIVKKAVRVSPDDDDKGFIEKEAPLHASKVRPICPACGKPTRVRKKFLENGKKIRVCAKCGGALDTEE

[0655] SEQ ID NO: 477MRVKMHVKKGDTVLVASGKYKGRVGKVKEVLPKKYAVIVEGVNIVKKAVRVSPEEEEEGFIEKEAPLHASKVRPICPACGKPTRVRKKFLENGKKIRVCAKCGGALDTEE

[0656] SEQ ID NO: 478MRVKMHVKKGDTVLVASGKYKGRVGKVKEVLPKKYAVIVEGVNIVKKAVRVSPRRRRRGFIEKEAPLHASKVRPICPACGKPTRVRKKFLENGKKIRVCAKCGGALDTEE

[0657] SEQ ID NO: 479MRVKMHVKKGDTVLVASGKYKGRVGKVKEVLPKKYAVIVEGVNIVKKAVRVSPCCCCCGFIEKEAPLHASKVRPICPACGKPTRVRKKFLENGKKIRVCAKCGGALDTEE

[0658] SEQ ID NO: 480MKLSEVRKQLEEARKLSPVELEKLVREKKRELMELRFQAS1GHHHHNHK1RDLKRQ1ARLLTVLNEKRRQNA

[0659] SEQ ID NO: 481MKLSEVRKQLEEARKLSPVELEKLVREKKRELMELRFQASIGHQHQNHKIRDLKRQIARLLTVLNEKRRQNA

[0660] SEQ ID NO: 482MKLSEVRKQLEEARKLSPVELEKLVREKKRELMELRFQASIGRRRRNHKIRDLKRQIARLLTVLNEKRRQNA

[0661] SEQ ID NO: 483MKLSEVRKQLEEARKLSPVELEKLVREKKRELMELRFQASIGEEEENHKIRDLKRQIARLLTVLNEKRRQNA

[0662] SEQ ID NO: 484MKLSEVRKQLEEARKLSPVELEKLVREKKRELMELRFQASIGCCCCNHKIRDLKRQIARLLTVLNEKRRQNA

[0663] SEQ ID NO: 485MAKHPVPKKKTSKARRDARRSHHALTPPTLVPCPHHHAMKPPHTVCPECGYYAGRKHHHH

[0664] SEQ ID NO: 486MAKHPVPKKKTSKARRDARRSHHALTPPTLVPCPHQHAMKPPHTVCPECGYYAGRKHQHQ

[0665] SEQ ID NO: 487MAKHPVPKKKTSKARRDARRSHHALTPPTLVPCPEEEAMKPPHTVCPECGYYAGRKEEEE

[0666] SEQ ID NO: 488MAKHPVPKKKTSKARRDARRSHHALTPPTLVPCPRRRAMKPPHTVCPECGYYAGRKRRRR

[0667] SEQ ID NO: 489MAKHPVPKKKTSKARRDARRSHHALTPPTLVPCPCCCAMKPPHTVCPECGYYAGRKCCCC

[0668] SEQ ID NO: 490MAKHPVPKKKTSKARRDARRSHHALTPPTLVPCPECKAMKPPHTVCPECGYYAGRKEPEA

Claims

What is claimed is:

1. A cell-free protein synthesis platform, comprising: a ribosome and a polymer membrane comprising a polymer membrane pore and an affinity binding site capable of binding the ribosome, wherein the polymer membrane separates a reaction compartment from a product compartment.

2. The cell-free protein synthesis platform of claim 1, wherein the ribosome is bound to the polymer membrane.

3. The cell-free protein synthesis platform of claim 2, wherein the ribosome is bound to the polymer membrane by a membrane bound protein.

4. The cell-free protein synthesis platform of claim 3, wherein the membrane bound protein is a translocon.

5. The cell-free protein synthesis platform of claim 3, wherein the membrane bound protein is a hinge protein.

6. The cell-free protein synthesis platform of claim 3, wherein the membrane bound protein is a translocon bound to a hinge protein.

7. The cell-free protein synthesis platform of claim 1, further comprising a reaction solution.

8. The cell-free protein synthesis platform of claim 7, wherein the reaction solution comprises a cell-free protein extract.

9. The cell-free protein synthesis platform of claim 8, wherein the cell-free protein extract comprises an excess of small ribosomal subunits.

10. The cell-free protein synthesis platform of claim 1, further comprising a housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compartment.

11. The cell-free protein synthesis platform of claim 1, further comprising a polymer membrane bag having an inside membrane surface and an outside membrane surface.

12. The cell-free protein synthesis platform of claim 11, wherein the affinity binding site is on the inside membrane surface of the polymer membrane bag.

13. The cell-free protein synthesis platform of claim 12, wherein the polymer membrane bag is a dialysis bag.

14. The cell-free protein synthesis platform of claim 1, wherein the ribosome is a modified ribosome.

15. The cell-free protein synthesis platform of claim 14, wherein the modified ribosome comprises one or more modified ribosome sites.

16. The cell-free protein synthesis platform of claim 14, wherein the modified ribosome comprises 1 to 10 ribosome ligand binding sites.

17. The cell-free protein synthesis platform of claim 14, wherein the modified ribosome comprises a ribosome ligand binding site.

18. The cell-free protein synthesis platform of claim 14, wherein the modified ribosome comprises two or more ribosome ligand binding sites.

19. The cell-free protein synthesis platform of claim 18, wherein the ribosome ligand binding sites are different ligand binding sites.

20. The cell-free protein synthesis platform of claim 17, wherein the ribosome ligand binding site is a molecular binding site.

21. The cell-free protein synthesis platform of claim 17, the ribosome ligand binding site is an antibody epitope.

22. The cell-free protein synthesis platform of claim 17, wherein the ribosome ligand binding site is selected from His-tag, His-Glu-tag, Asp-Lys-tag, Glu-tag, Arg-tag, Cys-tag, FLAG-tag, Capture Select C-tag, TC-tag, Xpress-tag, Softag-3-tag, Strep-tag, Aul, AU5, T7-tag, V5-tag, B- tag, E2 epitope, HSV epitope, KT3 epitope, Myc epitope, PDZ ligands, Asp-tag, Phe-tag, Protein C, Sl-tag, Universal-tag, VSV-G, a HTTPHH, functional equivalents, or combinations thereof.

23. The cell-free protein synthesis platform of claim 17, wherein the ribosome ligand binding site is on a ribosomal protein.

24. The cell-free protein synthesis platform of claim 23, wherein the ribosomal protein is from a different organism than the ribosome.

25. The cell-free protein synthesis platform of claim 23, wherein the ribosomal protein is modified from its native form.

26. The cell-free protein synthesis platform of claim 17, wherein the ribosome ligand binding site is on a ribosomal RNA.

27. The cell-free protein synthesis platform of claim 26, wherein the ribosomal RNA is from a different organism than the ribosome.

28. The cell-free protein synthesis platform of claim 26, wherein the ribosomal RNA is modified from its native form.

29. The cell-free protein synthesis platform of claim 1, wherein the affinity binding site comprises an affinity molecule.

30. The cell-free protein synthesis platform of claim 29, wherein the affinity molecule is capable of binding a ribosome ligand binding site.

31. The cell-free protein synthesis platform of claim 29, wherein the affinity binding site comprises an affinity molecule that binds a ribosome ligand binding site.

32. The cell-free protein synthesis platform of claim 1, wherein the ribosome is attached to the polymer membrane.

33. The cell-free protein synthesis platform of claim 29, wherein the ribosome is bound to one or more affinity molecules on the polymer membrane.

34. The cell-free protein synthesis platform of claim 29, wherein the ribosome is bound to 1 to 10 affinity molecules on the polymer membrane.

35. The cell-free protein synthesis platform of claim 29, wherein the ribosome is bound to 1-3, 1- 5, 1-10, 2-5, 2-10, 3-5, 3-10, 4-6, or 4-10 affinity molecules on the polymer membrane.

36. The cell-free protein synthesis platform of claim 1, wherein the ribosome contains a naturally occurring ribosome ligand binding site.

37. The cell-free protein synthesis platform of claim 36, wherein the ribosome ligand binding site is capable of binding an affinity molecule of claim 29.

38. The cell-free protein synthesis platform of claim 29, wherein the affinity molecule is bound near the polymer membrane pore.

39. The cell-free protein synthesis platform of claim 29, wherein the affinity molecule is bound on the polymer membrane pore edge.

40. The cell-free protein synthesis platform of claim 29, wherein the affinity molecule is 0.1-100 nm, 0.1-50 nm, 0.1-25 nm, 0.1-10 nm, 0.1-5 nm, 0.1-1 nm, 0.5-100 nm, 0.5-50 nm, 0.5-25 nm, 0.5 nm-10 nm, 0.5-5 nm, 1-100 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-50 nm, 5- 25 nm, or 5-10 nm from the polymer membrane pore edge.

41. The cell-free protein synthesis platform of claim 29, wherein the affinity molecule is selected from a metal chelating tag, an antibody epitope, a covalent binding molecule, and functional equivalents and combinations thereof.

42. The cell-free protein synthesis platform of claim 41, wherein the affinity molecule is selected from HAT-tag, His-tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp-tag), cation exchange (Arg-tag), antibody epitopes (AU1, AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, Phenyl-Sepharose (Phe-tag), ProC,SplAsH (Cys-tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA-aptamer, RNA- Sephadex, RNA-Streptavadin), Tags made by the SELEX process, or combinations thereof.

43. The cell-free protein synthesis platform of claim 29, wherein each polymer membrane pore has 1 to 10 affinity binding molecules within 100 nm of the edge of the polymer membrane pore.

44. The cell-free protein synthesis platform of claim 29, comprising two or more different affinity molecules.

45. The cell-free protein synthesis platform of claim 14, wherein the modified ribosome is attached to the polymer membrane.

46. The cell-free protein synthesis platform of claim 1, wherein the ribosome is a reconstituted ribosome, a recombinant ribosome, or a hybrid ribosome.

47. The cell-free protein synthesis platform of claim 29, wherein the ribosome ligand binding site is attached via one or more intermediate molecules to the affinity molecule attached to the polymer membrane.

48. The cell-free protein synthesis platform of claim 47, wherein the intermediate molecule is a signal recognition protein.

49. The cell-free protein synthesis platform of claim 48, wherein the signal recognition protein is selected from SEQ ID NO: 257 to SEQ ID NO: 271, functional equivalents, or combinations thereof.

50. The cell-free protein synthesis platform of claim 47, wherein the intermediate molecule is a polymer, antibody, antibody complex, small molecule chemical linker, dendrimer, hinge protein, or combinations thereof.

51. The cell-free protein synthesis platform of claim 47, wherein the ribosome ligand binding site is attached via 2-10 intermediate molecules to the affinity molecule on the polymer membrane.

52. The cell-free protein synthesis platform of claim 46, wherein the hybrid ribosome comprises molecules selected from two or more organisms.

53. The cell-free protein synthesis platform of claim 46, wherein the hybrid ribosome comprises molecules selected from archaea, eukaryotes, prokaryotes, or combinations thereof.

54. The cell-free protein synthesis platform of claim 1, wherein the ribosome comprises a modified ribosome protein.

55. The cell-free protein synthesis platform of claim 54, wherein the modified ribosome protein comprise modifications selected from amino acid substitution, protein tags, non-canonical amino acid substitution, or combinations thereof.

56. The cell-free protein synthesis platform of claim 55, wherein the modified ribosome protein is selected from SEQ ID NO:1 to 256, SEQ ID NO: 272 to SEQ ID NO:490, functional equivalents, or combinations thereof.

57. The cell-free protein synthesis platform of claim 17, wherein the ribosome ligand binding site is a naturally occurring ribosomal RNA sequence.

58. The cell-free protein synthesis platform of claim 17, wherein the ribosome ligand binding site is a modified ribosomal RNA sequence.

59. The cell-free protein synthesis platform of claim 29, wherein the affinity molecule is an RNA tag.

60. The cell-free protein synthesis platform of claim 59, wherein the RNA tag is selected from RNA-biotin, RNA-aptamer, RNA-Sephadex, RNA-Streptavadin, functional equivalents, or combinations thereof.

61. The cell-free protein synthesis platform of claim 29, wherein a ribosomal RNA ligand binding site binds the affinity molecule on a polymer membrane.

62. The cell-free protein synthesis platform of claim 29, wherein one or more ribosome ligand binding sites can bind one or more affinity molecules near a membrane pore.

63. The cell-free protein synthesis platform of claim 7, wherein the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts.

64. The cell-free protein synthesis platform of claim 63, wherein the reaction solution further comprises an excess of small ribosomal subunits.

65. The cell-free protein synthesis platform of claim 63, wherein the reaction solution further comprises an mRNA for a protein of interest.

66. The cell-free protein synthesis platform of claim 63, wherein cofactors in the cell free protein synthesis extract comprise proteins, ions, and RNA.

67. The cell-free protein synthesis platform of claim 63, wherein the tRNA’s comprise naturally occurring, modified and synthesized tRNA molecules.

68. The cell-free protein synthesis platform of claim 63, wherein amino acids are selected from naturally occurring amino acids, noncanonical amino acids, or combinations thereof.

69. The cell-free protein synthesis platform of claim 68, wherein the amino acids are selected from is selected from alanine, arginine, asparagine, aspartate, cysteine, glutamine, glutamate, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine, pryrrolysine (stop codon amino acids), homoallylglycine, homopropargylglycine, azidohomoalanine, azidonorleucine, azidophenylalanine, acetylphenylalanine, acetylphenylalanine, propargyloxyphenylalanine, functional equivalents, or combinations thereof.

70. The cell-free protein synthesis platform of claim 63, wherein the energy molecules comprise nucleoside triphosphates (NTPs).

71. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane is comprised of a monomers or polymers selected from acrylamide, 3-acrylamidopropyl trimethylammonium chloride, N-hydroxymethyl acrylamide. N,N-mcthylcncbisacrylamidc, cellulose acetate, N,N-dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n-butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, poly sacrylamido-2-methyl- 1 -propane- sulfonic acid, polyacrylic acid, polymethacrylic acid, polyisopropylacrylamide, poly-3- acrilamidopropyl trimcthylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polydiallyldimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide, isopropylacrylamide, or vinylpyrrolidone, copolymers of acrylamide-2-methyl- 1 propanesulfonicacid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido- propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl- pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidonc, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfide, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl- ammonium, polyvinyl chloride, polyvinylidene fluoride, poly-4-vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinylsulfonic acid, functional equivalents, and combinations thereof..

72. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane is a hydrogel.

73. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane has a thickness of at least 1 nm, 5 nm, 1 Onm, 15 nm, 20 nm, 25 nm, 30 nm, 50 nm, 75 nm, or 100 nm.

74. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane has a thickness selected from of 1-100 nm, 1-75 nm, 1-50 nm, 1-25 nm, 1-10 nm, 1-5 nm, 5-100 nm, 5-75 nm, 5-50 nm, 5-25 nm, 5-10 nm, 10-100 nm, 10-75 nm, 10-50 nm, 10-25 nm, 25-100 nm, 25-75nm, or 25-50nm.

75. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane has a thickness of 5 nm to 10 nm.

76. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane is comprised of one or more layers.

77. The cell-free protein synthesis platform of claim 76, wherein the polymer membrane is comprised of 1-2, 1-3, 1-5, 1-10, 2-5, 2-10, or 5-10 layers.

78. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane comprises one or more layers of a polymer support matrix.

79. The cell-free protein synthesis platform of claim 78, wherein the polymer membrane comprises 1-2, 1-3, 1-5, 1-10, 2-5, 2-10, or 5-10 layers of a polymer support matrix.

80. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane is a semi permeable membrane.

81. The cell-free protein synthesis platform of claim 1 , wherein the polymer membrane is selectively permeable to water and ions of interest.

82. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane pores have a diameter of about 3 nm to about 15 nm.

83. The cell-free protein synthesis platform of claim 62, wherein the polymer membrane pores have a diameter selected from 5-100 nm, 5-75 nm, 5-50 nm, 5-25 nm, 5-10 nm, 10-10 Onm, 10- 75 nm, 10-50 nm, 10-25 nm, 20-100 nm, 20-75 nm, 20-50 nm, 20-25 nm, 50-100 nm, or 50-75 nm.

84. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane pores are spaced at least 10 nm, 20 nm, 25 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, or 500 nm edge to edge apart from other membrane pores.

85. The cell-free protein synthesis platform of claim 29, wherein the polymer membrane comprises 1-10 affinity molecules within 10 nm of the edge of each membrane pore.

86. The cell-free protein synthesis platform of claim 29, wherein the polymer membrane comprises 1-5 affinity molecules within 10 nm of the edge of each membrane pore.

87. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane comprises one or more coatings.

88. The cell-free protein synthesis platform of claim 1, wherein the polymer membrane comprises one or more embedded membrane protein.

89. The cell-free protein synthesis platform of claim 88, wherein the embedded membrane protein is a translocon.

90. The cell-free protein synthesis platform of claim 89, wherein the translocon binds to a modified ribosome.

91. The cell-free protein synthesis platform of claim 89, wherein the translocon binds a hinge protein, intermediate molecule, dendrimer, antibody, antibody complex, or combinations thereof.

92. The cell-free protein synthesis platform of claim 14, wherein a hinge protein, intermediate molecule, dendrimer, antibody, antibody complex, or combinations thereof binds to the modified ribosome.

93. The cell-free protein synthesis platform of claim 88, wherein the one or more embedded membrane protein comprises a membrane protein pore channel that allows passage of a polypeptide chain.

94. The cell-free protein synthesis platform of claim 1, wherein the ribosome can translocate a protein through the pore of the membrane protein.

95. The cell-free protein synthesis platform of claim 10, wherein the housing comprises a container with one or more partitions separating the container into two or more chambers comprising a reaction chamber and a product chamber.

96. The cell-free protein synthesis platform of claim 95, wherein the housing comprises a container with 1-3, 1-5, or 1-10 partitions separating the container into 2-4, 2-6, or 2-11 separate chambers.

97. The cell-free protein synthesis system of claim 10, wherein the housing comprises the polymer membrane.

98. The cell-free protein synthesis platform of claim 95, wherein the one or more partitions comprise the polymer membrane.

99. The cell-free protein synthesis platform of claim 1, further comprising a membrane support structure.

100. The cell-free protein synthesis platform of claim 99, wherein the membrane support structure is a frame.

101. The cell-free protein synthesis platform of claim 100, wherein the frame is comprised of a polymer.

102. The cell-free protein synthesis platform of claim 101, wherein the polymer is comprised of monomers or polymers selected from acrylamide, 3-acrylamidopropyl trimethylammonium chloride, N-hydroxymethyl acrylamide, N,N-methylenebisacrylamide, cellulose acetate, N,N- dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n- butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, polysacrylamido-2-methyl-l -propane-sulfonic acid, polyacrylic acid, polymethacrylic acid, polyisopropylacrylamide, poly-3-acrilamidopropyl trimethylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polydially Idimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide, isopropylacrylamide, or vinylpyrrolidone, copolymers of acrylamide-2-methyl-l propanesulfonic acid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido-propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl-pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidone, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfide, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl-ammonium, polyvinyl chloride, poly vinylidene fluoride, poly-4-vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinylsulfonic acid, functional equivalents and combinations thereof.

103. The cell-free protein synthesis platform of claim 10, wherein the housing comprises a vessel.

104. The cell-free protein synthesis platform of claim 103, wherein the vessel is comprised of a polymer comprised of monomers or polymers selected from acrylamide, 3-acrylamidopropyl trimethylammonium chloride, N-hydroxymethyl acrylamide, N,N-methylenebisacrylamide, cellulose acetate, N,N-dimethylacrylamide, N-methacryloylarcylamide, N-methyl-N-vinyl acetamide, ethyl acrylate, n-butyl acrylate, dodecyl acrylate, octyl acrylate, propyl acrylate, octadecyl acrylamide, stearyl acrylate, polyacrylamide, poly sacry lamido-2-methyl- 1 -propane-sulfonic acid, polyacrylic acid, polymethacrylic acid, polyisopropylacrylamide, poly-3- acrilamidopropyl trimethylammonium, polyacrylonitrile, polyacrylonitrile butadiene styrene, polyamide, polybenzimidazole, polycarbonate, polydiallyldimethyl-ammonium, polyester, polyether sulfone, polyether ether ketone, polyetherimide, polyether sulfone, polyethylene, polyethylene glycol diacrylate, polyethylene glycol dimethyl acrylate, polyethyleneimine, polyethylene oxide, copolymers of acrylic acid or methacrylic acid with acrylamide, isopropylacrylamide, or vinyl pyrrolidone, copolymers of acrylamide-2-methyl- 1 propanesulfonic acid with acrylamide, isopropyl acrylamide, or vinylpyrrolidone, copolymers of (3-acrylamido- propyl)trimethylammonium chloride with acrylamide, isopropylacrylamide, or N-vinyl- pyrrolidone, and copolymers of diallyl dimethylammonium chloride with acrylamide, isopropylacrylamide, or vinylpyrrolidone, polyhydroxy methyl acrylate, polyimide, polyisobutylene, polylactic acid, polymethylmethacrylate, polyoxymethylene, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, styrene, polystyrene sulfonic acid, polysulfide, polytetrafluoroethylene, polyvinyl alcohol, polyvinyl benzyl-N-trimethyl- ammonium, polyvinyl chloride, polyvinylidene fluoride, poly-4- vinyl-N-methylpyridinium, polyvinylpyrrolidone, polyvinylsulfonic acid, functional equivalents and combinations thereof.

105. The cell-free protein synthesis platform of claim 102, wherein the vessel contains a membrane support structure.

106. The cell-free protein synthesis platform of claim 10, wherein the housing comprises a flow reactor.

107. The cell-free protein synthesis platform of claim 106, wherein the flow reactor comprises one or more inlet / outlet connections.

108. The cell-free protein synthesis platform of claim 106, wherein the flow reactor comprises one or more inlet / outlet connections for product.

109. The cell-free protein synthesis platform of claim 106, wherein the flow reactor comprises the polymer membrane.

110. The cell-free protein synthesis platform of claim 109, wherein the polymer membrane of the flow reactor separates the flow reactor into one or more partitions.

111. The cell-free protein synthesis platform of claim 106, wherein the ribosome translates a protein through the polymer membrane pore into a product reaction chamber of the flow reactor.

112. A cell-free protein synthesis platform comprising: a ribosome bound to a polymer membrane comprising an affinity binding site and a polymer membrane pore, wherein the polymer membrane separates a reaction compartment from a product compartment.

113. The cell-free protein synthesis platform of claim 112, wherein the ribosome is bound to the polymer membrane by a membrane bound protein.

114. The cell-free protein synthesis platform of claim 113, wherein the membrane bound protein is a translocon and hinge protein.

115. The cell-free protein synthesis platform of claim 114 comprising a hinge protein.

116. The cell-free protein synthesis platform of claim 113, wherein the membrane bound protein is a hinge protein.

117. A kit for cell-free protein synthesis comprising a polymer membrane which comprises an affinity tag and a pore, and a large ribosome subunit attached to the polymer membrane.

118. The kit for cell-free protein synthesis of claim 117 further comprising a reaction solution.

119. The kit for cell-free protein synthesis of claim 118, wherein the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts.

120. The kit for cell-free protein synthesis of claim 118, wherein the reaction solution comprises an excess of small ribosomal subunits.121 . The kit for cell-free protein synthesis of claim 116, further comprising an mRNA for a protein of interest.

122. A kit for cell-free protein synthesis comprising a dialysis bag which comprises a polymer membrane with an affinity tag and a pore, and large ribosomal subunit attached to the polymer membrane.

123. The kit for cell-free protein synthesis of claim 122 further comprising a reaction solution.

124. The kit for cell-free protein synthesis of claim 123, wherein the reaction solution comprises cofactors, tRNA’s, amino acids, energy molecules, buffers, salts.

125. The kit for cell-free protein synthesis of claim 122, wherein the reaction solution comprises an excess of small ribosomal subunits.

126. The kit for cell-free protein synthesis of claim 122, further comprising an mRNA for a protein of interest.

127. The kit for cell-free protein synthesis of 122, wherein the reaction solution is inside the dialysis bag.

128. The kit for cell-free protein synthesis of 127, further comprising an mRNA of a protein of interest.

129. A cell-free protein synthesis platform comprising: a large ribosome subunit, a polymer membrane comprising an affinity binding site and a polymer membrane pore, wherein the large ribosome subunit is bound to the polymer membrane and the polymer membrane separates a reaction compartment from a product compartment.

130. The cell-free protein synthesis platform of claim 129, wherein the large ribosome subunit is bound to the polymer membrane by a membrane bound protein.

131. The cell-free protein synthesis platform of claim 130, wherein the membrane bound protein is a translocon.

132. The cell-free protein synthesis platform of claim 130, wherein the membrane bound protein is a hinge protein.

133. The cell-free protein synthesis platform of claim 130, wherein the membrane bound protein is a translocon bound to a hinge protein.

134. The cell-free protein synthesis platform of claim 129, further comprising a reaction solution.

135. The cell-free protein synthesis platform of claim 134, wherein the reaction solution comprises a cell-free protein synthesis extract.

136. The cell-free protein synthesis platform of claim 134, wherein the reaction solution comprises an excess of ribosome small subunits.

137. The cell-free protein synthesis platform of claim 129, further comprising a housing with two or more compartments wherein the polymer membrane separates a housing reaction compartment from a housing product compartment.

138. The cell-free protein synthesis platform of claim 129, further comprising a polymer membrane bag having an inside membrane surface and an outside membrane surface.

139. The cell-free protein synthesis platform of claim 138, wherein the affinity binding site is on the inside membrane surface of the polymer membrane bag.

140. The cell-free protein synthesis platform of claim 138, wherein the polymer membrane bag is a dialysis bag.

141. The cell-free protein synthesis platform of claim 129, wherein the large ribosome subunit ribosome is modified at one or more sites.

142. The cell-free protein synthesis platform of claim 14, wherein the large ribosome subunit is modified at one or more sites comprises and comprises 1 to 10 ribosome ligand binding sites.

143. The cell-free protein synthesis platform of claim 71, further comprising dendrimers for controlling pore diameter and distribution.

144. The cell-free protein synthesis system of claim 143, wherein the dendrimers contain affinity molecules.

145. The cell-free protein synthesis system of claim 144, wherein the dendrimer affinity molecules are selected form HAT-tag, His-tag, (Ni, Zn, Cu, Co, Zi, Mn), anion exchange (Asp- tag), cation exchange (Arg-tag), antibody epitopes (AUl, AU5, B-tag, E2, EE, FLAG, HAV5, HSV, KT3, MYC, SI, T7, TRP-E, Universal [HTTPHH], VSV-G), calmodulin binding peptide (CBP), cellulose binding domain (CBD), PDZ ligand, PhenyLSepharose (Phe-tag), ProC, SplAsH (Cys-tag), RNase (S-tag), streptavadin (STREP-tag), RNA tags (RNA-biotin, RNA- aptamer, RNA-Sephadex, RNA-Streptavadin), tags made by the SELEX process, functional equivalents or combinations thereof.

146. The cell-free protein synthesis system of claim 143, wherein the dendrimer material is selected from carbosilane, citric acid, glycodendrimers, peptide, phosphorus, polyamido-amine, polyether, polyglycerol, poly L-lysine, polypropylene-imine, triazine, functional equivalents or combinations thereof.

147. The cell-free protein synthesis system of claim 1, wherein the product compartment further comprises a reaction chamber.

148. The cell-free protein synthesis system of claim 147, wherein the reaction chamber further comprises enzymes, substrates, and reagents.

149. The cell-free protein synthesis system of claim 148, wherein the enzyme are enzymes that assist in protein folding, post-translational modification, or combinations thereof.

150. The cell-free protein synthesis system of claim 149, wherein the enzymes that assist in protein folding are chaperone proteins.

151. The cell-free protein synthesis system of claim 150, wherein the chaperone proteins are chaperonins.

152. The cell-free protein synthesis system of claim 149, wherein enzymes are selected from Chaperonin 10, Chaperonin 60, DnaK, FIMC, GroEL, GroES, GroESL, hchA, HSCP 70, HSP32, HSP70, HSPD1, PSMG2, PSMG4, SKP, functional equivalents, and combinations thereof.

153. The cell-free protein synthesis system of claim 149, wherein enzymes that perform post- translational modification are enzymes that catalyze acetylation, disulfide bond formation, glutathionylation, glycation, glycosylation, hydroxylation, lipidation, methylation, phosphorylation, proteolysis, S-nitrosylation, succinylation, SUMOylation, trisulfide bond formation, ubiquitination, functional equivalents, and combinations thereof.

154. The cell-free protein synthesis system of claim 148, wherein the reagent is a macromolecular crowding reagent.

153. The cell-free protein synthesis system of claim 152, wherein macromolecular crowding reagents are selected from Dextran 70, Ficoll 70, PEG, colloid-polymer mixtures, liposomes, functional equivalents, and combinations thereof.

154. The cell-free protein synthesis system of claim 152, wherein liposomes are prepared from cationic lipids, fatty acids, glycerolipids, ionizable lipids, PEGylated lipids, phospholipids, sterols, functional equivalents, and combinations thereof.

155. The cell-free protein synthesis system of claim 147, wherein reaction chamber further comprises metal ions.

156. The cell-free protein synthesis system of claim 155, wherein metal ions are selected from Ca, Co, Cr, Cu, Fe, K, Mg, Mo, Mn, Na, Ni, V, Zn, functional equivalents, and combinations thereof.

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