Chimeric cannulae proteins and methods for making and using them

Abstract

A polymer is prepared by self-assembly of a plurality of monomeric polypeptide units. The polymer tends to form a nanotube and is capable of encapsulating a particular drug molecule. Once encapsulated in the polymer of the present invention, the drug molecule may be delivered to a particular location of human body to effectively cure a disease or treat a symptom. Generally, the monomeric polypeptide unit of the present invention has a sequence found in Pyrodictium abyssi , a microorganism that produces an extracellular network having hollow protein tubes, or a sequence substantially identical thereto. The monomeric polypeptide may be mass produced using recombinant biotechnologies and be polymerized into the polymer of the present invention. One or more additional targeting vector may be attached to the monomeric polypeptide unit or the polymer to facilitate the targeting of the drug molecule that may be held there within. The sequence contained in the monomeric polypeptide unit may be further optimized using one or more technique selected from Gene Site Saturation Mutagenesis and GeneReasembly.

1 . A polypeptide comprising, or consisting of:
 (a) (i) a polypeptide comprising an amino acid sequence having at least 70%, 80%, 90%, 95% or 99% or complete sequence identity to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, or SEQ ID NO:10; and   (ii) at least one functional group comprising, or consisting of: an antibody; an oligosaccharide, a polynucleotide (nucleic acid), a glycosylation, a polymer gel, a polyethylene glycol, a lipid or lipid derivative, a phosphytidylinositol, a flavin, a heme moiety, a vector, a morphatide, a derivatized nucleotide complex that can bind a specific antigen, a polynucleotide scaffold, or a combination thereof.   
     
     
         2 . The polypeptide of  claim 1 , wherein the at least one functional group comprises, or consists of, a polynucleotide (nucleic acid). 
     
     
         3 . The polypeptide of  claim 1 , wherein the at least one functional group comprises, or consists of, a polyethylene glycol. 
     
     
         4 . The polypeptide of  claim 1 , wherein the at least one functional group comprises, or consists of, an oligosaccharide. 
     
     
         5 - 24 . (canceled) 
     
     
         25 . The chimeric polypeptide of  claim 1 , wherein the at least one functional group comprises, or consists of, an antibody or a single-chain antibody. 
     
     
         26 . The polypeptide of  claim 1 , wherein all differences in amino acid residues between the polypeptide and SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, or SEQ ID NO:10, are conservative amino acid substitutions of one amino acid for another amino acid. 
     
     
         27 . The polypeptide of  claim 1 , wherein a plurality of polypeptides polymerize through a self-assembly process, a chain polymerization reaction and/or a step polymerization reaction. 
     
     
         28 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize through a self-assembly process in the presence of (a) at least one divalent cation; (b) at least one divalent cation selected from the group consisting of Ca 2+ , Mg 2+ , Cu 2+ , Zn 2+ , Sr 2+ , Ni 2+ , Mn 2+  and Fe 2+ ; (c) at least one divalent cation comprising a Ca 2+ ; (d) at least one divalent cation comprising Ca 2+  and Mg 2+ ; and/or (e) a template molecule and an alkaline earth metal ion. 
     
     
         29 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to form a nanoscale drug capsule or a nanoscale delivery vehicle. 
     
     
         30 . The polypeptide of  claim 29 , wherein the nanoscale drug capsule or nanoscale delivery vehicle is a nanocapsule, a nanotube, a nanoparticle, a nanocage and/or a micelle. 
     
     
         31 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to encapsulate a material. 
     
     
         32 . The polypeptide of  claim 31 , wherein the encapsulated material comprises a drug. 
     
     
         33 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to form a fiber. 
     
     
         34 . The polypeptide of  claim 1 , wherein the polypeptide of (a) (i) and the at least one functional group of (a) (ii) are joined by a linker group. 
     
     
         35 . The polypeptide of  claim 1 , wherein the at least one functional group of (ii) is a polypeptide and the polypeptide of (a) (i) and the polypeptide of (a) (ii) are joined as a recombinant fusion protein. 
     
     
         36 . The polypeptide of  claim 1 , wherein the polypeptide of (a) (i) and the at least one functional group of (a) (ii) are joined with each other by a covalent linkage, or by a pairing, bundling, entangling and/or electrostatic cross-linking or bridging by divalent cations. 
     
     
         37 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to form structures comprising a rod, a pair of rods or rods in an interconnected network. 
     
     
         38 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to form a biochip structure. 
     
     
         39 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to form a nanomachinery structure. 
     
     
         40 . The polypeptide of  claim 27 , wherein a plurality of polypeptides polymerize to form a high-density, three-dimensional format. 
     
     
         41 . The polypeptide of  claim 1 , further comprising a hydrogel, a polyethyleneoxide, a copolymer of polyethyleneoxide and polypropyleneoxide, a water-active diisocyanate or a polyol. 
     
     
         42 . A recombinant fusion polypeptide comprising, or consisting of:
 (a) (i) an amino acid sequence having at least 70%, 80%, 90%, 95% or 99% or complete sequence identity to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, or SEQ ID NO:10; and   (ii) an amino acid sequence comprising at least one functional group.   
     
     
         43 . The recombinant fusion polypeptide of  claim 42 , wherein the at least one functional group comprises, or consists of: an antibody. 
     
     
         44 . The recombinant fusion polypeptide of  claim 42 , wherein the at least one functional group of (a) (ii) further comprises an oligosaccharide, a polynucleotide (nucleic acid), a glycosylation, a polymer gel, a polyethylene glycol, a lipid or lipid derivative, a phosphytidylinositol, a flavin, a heme moiety, a vector, a morphatide, a derivatized nucleotide complex that can bind a specific antigen, a polynucleotide scaffold, or a combination thereof. 
     
     
         45 . The recombinant fusion polypeptide of  claim 44 , wherein the at least one functional group of (a) (ii) is joined with the oligosaccharide, polynucleotide (nucleic acid), glycosylation, polymer gel, polyethylene glycol, lipid or lipid derivative, phosphytidylinositol, flavin, heme moiety, vector, morphatide, derivatized nucleotide complex that can bind a specific antigen and/or polynucleotide scaffold by a covalent linkage, or a pairing, bundling, entangling and/or electrostatic cross-linking or bridging by divalent cations.