Wet-spun Ramin-based fibers

Abstract

A fiber comprising a lamin system protein, the fiber being characterized by a diameter of 10 μm to 180 μm. Additionally, articles comprising the fiber and methods of making the same are disclosed.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 245,910, filed September 19, 2021, the contents of which are incorporated herein by reference in their entirety.

[0002] The present invention relates generally to the field of Ramin-based fibers. [Background technology]

[0003] Proteinaceous fibers are ubiquitous in biological systems and exist in versatile structures that exhibit a wide range of mechanical properties, from stiff to elastic, to address diverse functional demands. Fibrous proteins have unique physical properties that have been exploited to design novel biomaterials such as scaffolds and fibers. In general, these protein fibers have high biocompatibility, making them ideal for tissue engineering, wound dressing, and drug delivery applications. Natural and regenerated silk from the silkworm (Bombyx mori) as well as spider dragline silk silk protein-based fibers have been rigorously studied due to their toughness and stiffness, which stand out from those of most natural protein-based fibers and synthetic materials. High stiffness and toughness are also observed in the micrometer-sized native silks (1-2 μm in diameter) of hagfish mucus, which are composed of "keratin-like" proteins (α and γ). Hagfish silks have been successfully spun from recombinantly expressed proteins, but the mechanical properties of these fibers were inferior to those of natural fibers. These findings suggest that to achieve high toughness and stiffness, the unique organization of proteins as found in natural fibers must be faithfully mimicked. Summary of the Invention [Problem to be solved by the invention]

[0004] In one aspect of the present invention, there is provided a fiber comprising a lamin system protein, characterized by a diameter of 10 μm to 180 μm.

[0005] In some embodiments, the lamin system protein comprises an A-type lamin, a B-type lamin, or both.

[0006] In some embodiments, the B-type lamin comprises the amino acid sequence set forth in SEQ ID NO:1, wherein X1 comprises Gln or Lys (including any functional analog having at least 70% sequence homology thereto).

[0007] In some embodiments, the fiber further comprises either: (i) an N-terminal region comprising the amino acid sequence set forth in SEQ ID NO:2 (including any functional analog having at least 70% sequence homology thereto); and (ii) a C-terminal region comprising the amino acid sequence set forth in SEQ ID NO:3 (including any functional analog having at least 70% sequence homology thereto).

[0008] In some embodiments, the A-type lamin comprises the amino acid sequence set forth in SEQ ID NO:4, wherein X2 is Glu or Lys (including any functional analog having at least 70% sequence homology thereto).

[0009] In another aspect of the invention, there is provided a fiber comprising an A-type lamin system protein, the fiber being characterized by a diameter of between 10 μm and 180 μm.

[0010] In some embodiments, the repeat region comprises the amino acid sequence set forth in SEQ ID NO:4, where X2 is Glu or Lys (including any functional analog having at least 70% sequence homology thereto).

[0011] In some embodiments, the amino acid sequence further comprises either: (i) an N-terminal region comprising the amino acid sequence set forth in SEQ ID NO:5; and (ii) a C-terminal region comprising the amino acid sequence set forth in SEQ ID NO:6.

[0012] In some embodiments, the fibers further comprise a B-type lamin.

[0013] In some embodiments, the fibers have the following: yield strength between 1 MPa and 1000 MPa; tensile strength between -1 MPa and 1000 MPa; strain at break between 10% and 500%; and 30 MJ / m 3 ~1000MJ / m 3 and a Young's modulus of 0.001 GPa to 30 GPa.

[0014] In some embodiments, the protein is characterized by a 3%-50% α-helix to β-sheet transition during its extension.

[0015] In some embodiments, the fibers are characterized by a diameter of between 20 μm and 80 μm.

[0016] In another aspect of the invention, there is provided a fiber comprising a B-type lamin system protein, the fiber being characterized by (i) a diameter of between 10 μm and 180 μm; (ii) the B-type lamin system protein comprises an amino acid set forth in SEQ ID NO:1 (including any functional analog having at least 70% sequence homology thereto), and when X1 is Gln, the fiber has (i) a residual amount of alcohol; (ii) 190 MJ / m 3 and (ii) the protein is characterized by a 3%-50% α-helix to β-sheet transition upon its extension.

[0017] In some embodiments, the fibers are characterized by a diameter of 30 to 60 μm.

[0018] In some embodiments, the Lamin system protein further comprises either: i. an N-terminal region comprising the amino acid sequence SEQ ID NO:2 or any functional analog having at least 70% sequence homology thereto; ii. a C-terminal region comprising the amino acid sequence SEQ ID NO:3 or any functional analog having at least 70% sequence homology thereto.

[0019] In some embodiments, the fibers have the following: yield strength between 1 MPa and 1000 MPa; tensile strength between -1 MPa and 1000 MPa; strain at break between 10% and 500%; and 30 MJ / m 3 ~1000MJ / m 3 and a Young's modulus of 0.001 GPa to 30 GPa.

[0020] In some embodiments, each repeat independently has a molecular weight in the range of 20 kDa to 80 kDa.

[0021] In some embodiments, the fibers are elongated fibers.

[0022] In some embodiments, the fibers comprise a plurality of lamin system proteins arranged in the form of paracrystals, each of the paracrystals being characterized by a dimension selected from: (i) a width between 1 nm and 500 nm; (ii) a length between 0.5 mm and 1 cm, or both (i) and (ii).

[0023] In some embodiments, the lamin system protein is an isolated protein.

[0024] In some embodiments, the fibers are obtained by expression in bacteria.

[0025] In some embodiments, the protein is characterized by a secondary structure comprising an alpha helix:beta sheet ratio of 5:1 to 1:1.

[0026] In some embodiments, the fiber further comprises a hydrophobic coating.

[0027] In some embodiments, the hydrophobic coating comprises vegetable oils, mineral oils, fatty acids, isobutyl stearate, tallow fatty acid 2-ethylhexyl esters, polyol carboxylic acid esters, coconut fatty acid esters of glycerol, alkoxylated glycerol, silicones, dimethylpolysiloxanes, polyalkylene glycols, polyethylene oxides, propylene oxide copolymers, or any combination thereof.

[0028] In another aspect of the invention, an article is provided comprising the fiber of the invention.

[0029] In some embodiments, the article is in the form of a woven or nonwoven substrate.

[0030] In some embodiments, the article is characterized by at least one improved mechanical property compared to the properties of an article not containing the fibers, the property being selected from the group consisting of Young's modulus, tensile strength, strain at break, yield point, toughness, work to failure, impact strength, tear strength, flexural modulus, flexural strain, and stress at a particular elongation.

[0031] In another aspect of the present invention, there is provided a method for obtaining fibers of the present invention, comprising the steps of: a. providing a Lamin system protein at a concentration of 10 mg / mL to 400 mg / mL; and b. injecting the Lamin system protein into a coagulation solution, thereby forming fibers.

[0032] In some embodiments, the coagulation solution is characterized by a viscosity of between 0.45 cP and 3 cP, or greater than 0.7 cP.

[0033] In some embodiments, the infusion is performed at a flow rate of at least 0.1 ml / h.

[0034] In some embodiments, the coagulation solution comprises (i) an alcohol, (ii) an aqueous solution, (ii) a buffer, or any combination thereof.

[0035] In some embodiments, the alcohol is selected from methanol (MeOH), ethanol (EtOH), propanol (PrOH), isopropyl alcohol (IPA), or any combination thereof.

[0036] In some embodiments, the coagulation solution comprises between 50% (v / v) and 100% (v / v) alcohol.

[0037] In some embodiments, the coagulation solution includes a cross-linking agent.

[0038] In some embodiments, the coagulation solution contains between 1 mM and 100 mM CaCl2.

[0039] In some embodiments, the method further comprises at least one of the steps of (i) drying the fibers, and (ii) stretching the fibers.

[0040] In some embodiments, the method further comprises the step (c) of contacting the fibers with a hydrophobic agent, thereby forming a coating layer on the fibers.

[0041] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used to carry out or test embodiments of the present invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, shall prevail. Furthermore, the materials, methods, and examples are illustrative only and are not necessarily intended to be limiting.

[0042] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. [Brief description of the drawings]

[0043] [Figure 1] Figure 1A-1H show the Ce-lamin fiber structure (Figure 1A), where the full-length-Ce-lamin gene is divided into three structural domains: head, rod, and tail. The rod domain contains three α-helical coiled-coil segments (1A, 1B, and 2) connected by L1 and L12 linkers. The majority of the tail domain (105 aa) folds into an immunoglobulin (Ig)-like globular structure. Deletion of the entire tail domain or both the head and tail domains resulted in rod-Ce-lamin and rod-tail-Ce-lamin constructs; schematic diagram of recombinant Ce-lamin produced in E. coli (Figure 1B) as inclusion bodies solubilized in urea buffer. Fibers were fabricated by wet spinning of an aqueous Ce-Lamin solution into a Ca+2 ion-containing aqueous solution; and SEM images of rod (Fig. 1C), rod-tail (Fig. 1D), and full-length (Fig. 1E) Ce-Lamin fibers before and after tensile testing, showing their fiber morphology, as well as higher magnification SEM images of rod (Fig. 1F), rod-tail (Fig. 1G), and full-length (Fig. 1H) Ce-Lamin fibers formed under optimal assembly conditions. Insets are lower magnification views.

[0044] [Diagram 2]Figures 2A-2C show the internal structure of Ce-lamin-based fibers: TEM images of 70 nm thick cross sections of rod-, rod-tail-, and full-length Ce-lamin-based wet fibers spun under optimal assembly conditions (Figure 2A). A series of images with gradually increasing magnification in the inset shows that the Ce-lamin fiber internal structure is a complex network of quasicrystals. Cross-sectional analysis of rod-, rod-tail-, and full-length Ce-lamin-based wet fibers imaged using TEM (Figure 2B): The structural diversity of the fibers was detected as the injection rate increased, 0.5, 1, and 3.5 mL / h, into a buffer containing 20 mM or 50 mM CaCl2. Most fibers contained a random network of quasicrystals exhibiting a wide range of diameters (Table 4). Electron micrograph of quasicrystals assembled from full-length Ce-lamin (Figure 2C). The average repeat length (dark / black and light / white segments), i.e., the length between two centers of the black regions, was measured as shown.

[0045] [Diagram 3] Graph of stress-strain curves of rod-, rod-tail, and full-length-Ce-lamin fibers at optimal assembly conditions; under these conditions, the three constructs demonstrated optimal mechanical properties.

[0046] [Figure 4] Figures 4A-4D are bar graphs of the effect of CaCl concentration in the coagulation bath, injection flow rate, and crosshead speed on the diameter, stiffness, and toughness of rod-Ce-Lamin-based fibers (Figure 4A), rod-tail-Ce-Lamin-based fibers (Figure 4B), and full-length Ce-Lamin-based fibers (Figure 4C). The crosshead speeds were 0.3 mm / min (left bar), 10 mm / min (middle bar), and 100 mm / min (right bar), and SEM images (Figure 4D) of full-length Ce-Lamin fibers formed at an injection rate of 1 mL / h in a coagulation bath containing 50 mM CaCl. Images were taken after tensile testing.

[0047] [Diagram 5]Figures 5A-5C show secondary structure analysis by Raman spectroscopy: Experimental Raman spectra of rod-Ce-lamin based fibers (Figure 5A), rod-tail-Ce-lamin based fibers (Figure 5B), and full-length Ce-lamin based fibers (Figure 5C) described in Figure 3 show peak shifts before and after mechanical testing. Peaks are found in the amide I band region, corresponding to α-helix (1650 cm-1) and β-sheet / random coil (1667 cm-1) structures. Graphical representation of the assessment of the relative increase in β-sheet / random coil structures performed by the EM algorithm statistical method (middle panel).

[0048] [Figure 6] Figures 6A-6B show a structural model of Ce-lamin fiber assembly: The basic building block of Ce-lamin is a coiled-coil dimer. The dimers polymerize in a head-to-tail manner to form a polymer of dimers, which then laterally associate to form tetrameric protofilaments. Protofilaments are probably the fundamental building blocks of all higher-order lamin structures in vitro and in cell nuclei. The next step is the association of protofilaments into quasicrystals, whose interconnections form a random network that constitutes the internal structure of Ce-lamin fibers (Figure 6A). In response to stretching, the quasicrystals / protofilaments are reorganized and aligned along the long axis of the fiber (Figure 6B).

[0049] [Figure 7] Figures 7A-7B are graphs depicting the α-β transition propagation during tensile test cessation at different strains, shown on typical stress-strain curves for full-length Ce-lamin fibers assembled in 70% ethanol (Figure 7A) and Ce-lamin fibers assembled in aqueous buffer containing 20 m m CaCl (Figure 7B).

[0050] [Figure 8] Raman spectroscopic curve analysis of full-length Ce-lamin fibers assembled in 70% ethanol.

[0051] [Figure 9]9A-9D are SEM images showing dried Ce-lamin fibers assembled in alcohol solutions prior to tensile testing: fibers formed in 70% ethanol (FIG. 9A), fibers formed in 50% ethanol (FIG. 9B), fibers formed in 70% IPA (FIG. 9C), and fibers formed in 50% IPA (FIG. 9D).

[0052] [Figure 10] 10A-10C are SEM images showing dried Ce-lamin fibers assembled in alcohol solutions after tensile testing: a fiber formed in 70% ethanol and stretched to 30% strain (FIG. 10A), a fiber formed in 70% IPA and stretched to 30% strain (FIG. 10B), and a fiber formed in 70% ethanol and stretched to failure (FIG. 10C).

[0053] [Figure 11] 1 is a graph of the stress-strain curve of Q159K-Ce-Lamin fibers assembled in an alcohol solution in water (Et-ethanol, IPA-isopropanol).

[0054] [Figure 12] Figures 12A-12D are graphs and Raman spectra showing the mechanical and structural behavior in the elastic region of Ce-Lamin fibers assembled in aqueous buffer containing 20 m m CaCl2 (12A, 12C) or 70% ethanol (12B, 12D) during 5 cycle strain testing. Figures 12A-B show graphs of the mechanical behavior of Ce-Lamin fibers assembled in aqueous buffer containing 20 m m CaCl2 (12A), and 70% ethanol (12B). Figures 12C-12D show Raman spectra of Ce-Lamin fibers assembled in aqueous buffer containing 20 m m CaCl2 (12C), and 70% ethanol (12D). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0055] According to some embodiments, the invention provides fibers comprising a lamin-based protein, characterized by an average diameter between 1 μm and 1000 μm, and further characterized by an average length of at least 0.1 cm. In some embodiments, the fibers are lamin-based fibers, which are obtained via a wet spinning process. The inventors have observed that wet spun fibers formed by injecting a lamin-based protein into a coagulation bath are characterized by a sufficient length to result in fibers with superior mechanical properties compared to similar self-assembled fibers. The invention is based, in part, on the surprising finding that such wet spun fibers are characterized by a toughness and stiffness comparable to natural dragline spider silk fibers and natural hagfish mucus silk.

[0056] Furthermore, the present invention is based in part on the surprising finding that fibers injected into an alcoholic coagulation solution are characterized by superior mechanical strength and improved elasticity compared to fibers injected into a coagulation solution. Such fibers were further characterized by specific Raman patterns, as disclosed below. The inventors hypothesized that the superior mechanical properties of fibers obtained from alcoholic coagulation are related to the α-helix to β-sheet transition pattern upon stretching the fibers, as characterized by Raman spectroscopy (see, e.g., FIG. 7A). The α-helix to β-sheet transition (e.g., at least 20%) occurs already in the elastic region (e.g., about 6% strain) in fibers obtained from alcoholic coagulation, while fibers obtained from a coagulation solution showed the α-helix to β-sheet transition only when fully stretched (strains of about 100% or more, see further FIG. 7B).

[0057] Moreover, the present invention is based, in part, on the surprising discovery that coated fibers of the present invention (e.g., including a hydrophobic coating) exhibit significantly improved mechanical strength compared to similar uncoated fibers.

[0058] According to some embodiments, the present invention provides a method for obtaining fibers comprising a Lamin system protein, the fibers being characterized by a diameter between 10 μm and 180 μm, the method comprising the steps of providing a Lamin system protein at a concentration between 10 mg / mL and 400 mg / mL, and injecting the Lamin system protein into a coagulation solution, thereby forming fibers.

[0059] The present invention is based, in part, on the discovery that the concentration of lamin system proteins is important for the formation of fibers having desired structure, thickness, and mechanical properties.

[0060] Ramin Fiber According to an aspect of some embodiments of the present invention, there is provided a fiber comprising a lamin system protein. In some embodiments, the fiber is a lamin system protein fiber. In some embodiments, the fiber is a lamin system protein fiber. In some embodiments, the fiber is a fiber having a diameter of 1 μm to 1000 μm, 1 μm to 200 μm, 1 μm to 500 μm, 1 μm to 300 μm, 1 μm to 400 μm, 1 μm to 700 μm, 50 μm to 1000 μm, 50 μm to 500 μm, 50 μm to 300 μm, 50 μm to 200 μm, 2 μm to 200 μm, 3 μm to 400 μm, ...50 μm to 300 μm, 50 μm to 200 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 300 μm, 50 μm to 30 m~200μm, 5μm~200μm, 7μm~200μm, 9μm~200μm, 1μm~190μm, 2μm~190μm, 3μm~190μm , 5μm~190μm, 7μm~190μm, 9μm~190μm, 10μm~180μm, 15μm~180μm, 20μm~180μm, 25μm ~180μm, 30μm~180μm, 50μm~180μm, 65μm~180μm, 10μm~100μm, 15μm~100μm, 20μm~ 100μm, 25μm~100μm, 30μm~100μm, 50μm~100μm, 65μm~100μm, 10μm~80μm, 15μm~80μ The fiber may be characterized by a diameter of 100 μm, 20 μm to 80 μm, 25 μm to 80 μm, 30 μm to 80 μm, 50 μm to 80 μm, 65 μm to 80 μm, 10 μm to 50 μm, 15 μm to 50 μm, 20 μm to 50 μm, 25 μm to 50 μm, or 30 μm to 50 μm (including any range therebetween). Each possibility represents a separate embodiment of the present invention. As used herein, the term "diameter" refers to the average cross-section of the dry fiber. The cross-section of the fiber may be determined by TEM, SEM or optical microscopy.

[0061] In some embodiments, the fibers are characterized by a uniform diameter distribution. In some embodiments, the fibers are characterized by a diameter distribution (SD) of 0.1-1.5, 0.2-1.5, 0.3-1.5, 0.5-1.5, 0.1-1.0, 0.2-1.0, 0.3-1.0, 0.5-1.0 (including any range therebetween). Each possibility represents a separate embodiment of the present invention. In some embodiments, the diameter distribution is quantified by the standard deviation (SD).

[0062] In some embodiments, the fibers are characterized by a length of at least 0.01 cm, at least 0.1 cm, at least 1 cm, at least 2 cm, at least 5 cm, at least 10 cm, at least 50 cm, at least 100 cm, at least 500 cm, at least 1000 cm, or at least 10,000 cm, 0.01-10,000 cm, 0.01-100.000 cm, 0.05-10.000 cm (including any value therebetween). Each possibility represents a separate embodiment of the present invention. As used herein, the term "length" refers to the average length of the dry fibers.

[0063] In some embodiments, the lamin system protein comprises an A-type lamin, a B-type lamin, or both.

[0064] As used herein, "lamin" refers to a fibrous protein in V-type intermediate filaments (IFs) that provides structural function and transcriptional regulation in the cell nucleus. Like all IF proteins, lamins have a tripartite structure consisting of a globular amino-terminal (head) domain, a carboxy-terminal (tail) domain, and a long α-helical domain flanked by a central α-helical or rod domain. Lamins are classified as A-type and B-type. B-type lamins are expressed in most cell types, whereas A-type lamins, including lamin C, are generally expressed in differentiated tissues.

[0065] The terms "fiber" and "filament" are used interchangeably to refer to a thin cord of textile material. "Filament" means a thin, elongated thread-like body or structure of indefinite length, ranging from microscopic lengths to lengths of a mile or more. In some embodiments, the term "fiber" refers to the basic structure of textile material, which cannot be further divided into smaller parts by using conventional industrial means (except harsh chemical conditions that result in denaturation and / or decomposition of the paracrystals or protofilaments that make up the fiber). In contrast to yarns (composed of two or more twisted or intertwined filaments), fibers encompass a basic structure that is not formed by multiple intertwined filaments. The fibers of the present invention encompass continuous lamin-based fibers that include multiple lamin-based proteins assembled in a paracrystal form. The paracrystals in the fibers of the present invention are aligned substantially perpendicularly along the longitudinal axis of the fiber. In some embodiments, according to the present invention, the filament is a biofilament. By "biofilament" is meant a filament made from proteins. In some embodiments, the fibers or filaments of the present invention are in a crystalline state. In some embodiments, the fibers or filaments of the present invention are in a paracrystalline state or are paracrystalline fibers. In some embodiments, the fibers of the present invention form a paracrystalline material. In some embodiments, the fibers or filaments of the present invention consist essentially of paracrystalline repeat units.

[0066] In some embodiments, the average distance between two adjacent quasicrystals (also referred to herein as "repeat length") is 20 to 60 nm, about 30 to about 50 nm, about 35 to about 50 nm, about 30 to about 40 nm, about 38 to about 45 nm (including any range or value therebetween). In some embodiments, the average distance between two adjacent quasicrystals is about 40 nm. The average distance can be determined by measuring the length between the centers of two adjacent dark regions based on a TEM image of the fiber (as demonstrated in FIG. 2C).

[0067] In some embodiments, the fibers or filaments of the invention are obtained via a wet spinning process (e.g., upon coagulation as described herein). In some embodiments, the fibers or filaments of the invention are wet spun fibers or consist essentially of wet spun fibers. In some embodiments, the fibers or filaments of the invention lack self-assembled Lamin fibers. The term "self-assembled fibers" as used herein encompasses fibers formed by the self-assembly of Lamin proteins into a protein network (e.g., by introducing isolated proteins into a CaCl2 solution) composed of microfibers, which are further wet spun into yarns. In some embodiments, the self-assembled Lamin fibers are in the form of yarns composed of microfibers characterized by an average length of less than 1 μm, or less than 1.5 μm, or between 300 and 1500 nm.

[0068] In some embodiments, a plurality of fibers or filaments of the invention are provided that are characterized by an average length of at least 1 cm, at least 10 cm, at least 100 cm, at least 1000 cm, at least 100,000 cm, or more (including any range or value therebetween). In some embodiments, a plurality of fibers or filaments of the invention are characterized by an average length of 0.1 cm to 10 cm, 0.1 cm to 100 cm, 0.1 cm to 1000 cm, 0.1 cm to 10,000 cm, 0.1 cm to 1 cm, 0.1 cm to 5 cm, 0.1 cm to 50 cm, 1 cm to 10 cm, 1 cm to 100 cm, 1 cm to 1000 cm, 1 cm to 10,000 cm, 10 cm to 100 cm, 10 cm to 1000 cm, 10 cm to 10,000 cm (including any range or value therebetween). In some embodiments, the fibers or filaments of the invention are substantially free of microfibers.

[0069] In some embodiments, the Lamin system protein fibers of the invention consist essentially of Lamin system proteins.

[0070] As used herein, the terms "polypeptide", "peptide" and "protein" are used interchangeably to refer to two or more amino acids linked together. As used herein, the terms "polypeptide", "peptide", "protein" and "amino acid sequence" refer to any compound that includes naturally occurring or synthetic amino acid polymers or amino acid-like molecules, including but not limited to compounds that include amino and / or imino molecules. No particular size is implied by the use of the terms "peptide", "oligopeptide", "polypeptide", or "protein". Included within the definition are, for example, polypeptides that contain one or more analogs of amino acids (including, for example, unnatural amino acids, etc.), polypeptides with substituted bonds, and other modifications known in the art, whether naturally occurring or non-naturally occurring (e.g., synthetic). Thus, included within the definition are synthetic oligopeptides, dimers, multimers (e.g., tandem repeats, multiple antigenic peptide (MAP) forms, linearly linked peptides), cyclized molecules, branched molecules, and the like. In another embodiment, the described peptides, polypeptides and proteins have modifications that make them more stable while in an organism or capable of penetrating into cells. In one embodiment, the terms "peptide," "polypeptide," and "protein" refer to naturally occurring amino acid polymers. In another embodiment, the terms "peptide," "polypeptide," and "protein" refer to amino acid polymers in which one or more amino acid residues are artificial chemical analogues of a corresponding naturally occurring amino acid.

[0071] In some embodiments, the lamin system protein comprises a type B lamin. In some embodiments, a type B lamin has the amino acid sequence: LQEKDHLTSLNSRLATYIDKVRQLEQENNRLQVQIRDIEVVEKKEKSNLADRFEAEKARLRRALDSAQDELAKYRIEYDAAKVEVKKLKPQVEKLERELAGAEEQALHAQSIADQS(X1)AKQKTLQARNDKLVVENDDLKKQNITLRDTVEGLKKAVEDETLLRTAANNKIKALEEDLAFALQQHKGEL EEVRHKRQVDMTTYAKQINDEYQSKLQDQIEEMRAQFKNNLHQNKTAFEDAYKNKLNAARERQEEAVSEAIHLRARVRDLETSSSGNASLIERLRSELDTLKRSFQEKLDDKDARIAELNQEIERMMSEFHDLLDVKIQLDAELKTYQALLE (SEQ ID NO: 1) (including any functional analog having at least 70% sequence homology thereto). In some embodiments, X1 comprises Gln or Lys.

[0072] In some embodiments, a B-type lamin has the amino acid sequence: LQEKDHLTSLNSRLATYIDKVRQLEQENNRLQVQIRDIEVVEKKEKSNLADRFEAEKARLRRALDSAQDELAKYRIEYDAAKVEVKKLKPQVEKLERELAGAEEQALHAQSIADQS(X1)AKQKTLQARNDKLVVENDDLKKQNITLRDTVEGLKKAVEDETLLRTAANNKIKALEEDLAFALQQHKGELEEV HKRQVDMTTYAKQINDEYQSKLQDQIEEMRAQFKNNLHQNKTAFEDAYKNKLNAARERQEEAVSEAIHLRARVRDLETSSSGNASLIERLRSELDTLKRSFQEKLDDKDARIAELNQEIERMMSEFHDLLDVKIQLDAELKTYQALLEGEEERL (SEQ ID NO: 11) (including any functional analog having at least 70% sequence homology thereto). In some embodiments, X1 comprises Gln or Lys.

[0073] In some embodiments, a B-type lamin comprises the amino acid sequence: (SEQ ID NO: 12) (including any functional analog having at least 70% sequence homology thereto).

[0074] In some embodiments, the functional analog comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 89%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, or 100% homology or identity with SEQ ID NO:1, or with SEQ ID NO:10, or with SEQ ID NO:11, or with SEQ ID NO:12. Each possibility represents a separate embodiment of the invention. In some embodiments, homologs of the repeat region of B-type lamin protein are provided that share at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO:1, or with SEQ ID NO:10, or with SEQ ID NO:11, or with SEQ ID NO:12. Each possibility represents a separate embodiment of the present invention.

[0075] The terms "homology" or "identity", used interchangeably herein, refer to the sequence identity between two amino acid sequences or two nucleic acid sequences, with identity being the more strict comparison. The phrases "percent identity or homology" and "% identity or homology" refer to the percentage of sequence identity found in a comparison of two or more amino acid sequences or nucleic acid sequences. The two or more sequences can be anywhere from 0 to 100% identical, or any value in between. Identity can be determined by comparing positions in each sequence that can be aligned for comparison to a reference sequence. If a position in the compared sequences is occupied by the same nucleotide base or amino acid, then the molecules are identical at that position. The degree of identity of amino acid sequences is a function of the number of identical amino acids at positions shared by the amino acid sequences. The degree of identity between nucleic acid sequences is a function of the number of identical or matching nucleotides at positions shared by the nucleic acid sequences. The degree of homology of amino acid sequences is a function of the number of amino acids at positions shared by the polypeptide sequences.

[0076] The following is a non-limiting example for calculating the homology or sequence identity (these terms are used interchangeably herein) between two sequences. The sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced into one or both of the first and second amino acid or nucleic acid sequences for optimal alignment, and non-homologous sequences can be ignored for comparison purposes). The optimal alignment is determined as the best score using the GAP program of the GCG software package with a Blossum 62 score matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5. The amino acid residues or nucleotides at the corresponding amino acid positions or nucleotide positions are then compared. If a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position. The percent identity between two sequences is a function of the number of identical positions shared by the sequences.

[0077] In some embodiments, the percent homology or identity described herein is calculated or determined using the Basic Local Alignment Search Tool (BLAST). In some embodiments, the percent homology or identity described herein is calculated or determined using the Blossum 62 scoring matrix.

[0078] The term "nucleic acid" is well known in the art. As used herein, "nucleic acid" generally refers to any molecule (e.g., a chain) of DNA, RNA or derivatives or analogs thereof that contain nucleotides. Nucleotides are composed of a nucleoside and a phosphate group. The nitrogenous base of a nucleoside includes naturally occurring purine or pyrimidine nucleosides found, for example, in DNA (e.g., adenine "A", guanine "G", thymine "T" or cytosine "C") or RNA (e.g., A, G, uracil "U" or C).

[0079] The term "nucleic acid molecule" includes, but is not limited to, single-stranded RNA (ssRNA), double-stranded RNA (dsRNA), single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), small RNA, circular nucleic acids, fragments of genomic DNA or RNA, degraded nucleic acids, amplification products, modified nucleic acids, plasmids or organelle nucleic acids, and artificial nucleic acids such as oligonucleotides.

[0080] In some embodiments, n is an integer from 2 to 100, 3 to 100, 5 to 100, 10 to 100, 20 to 100, 30 to 100, 35 to 100, 40 to 100, 50 to 100, 2 to 80, 3 to 80, 5 to 80, 10 to 80, 20 to 80, 30 to 80, 35 to 80, 40 to 80, 50 to 80, 2 to 10, 3 to 6, 5 to 8, or 10 to 15 (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0081] In some embodiments, the lamin system protein (e.g., any one of SEQ ID NO:1, SEQ ID NO:11) further comprises an N-terminal region comprising the amino acid sequence: MSSRKGTRSSRIVTLERSANSSLSNNGGGDDSFGSTLLETSR (SEQ ID NO:2) (including any functional analog having at least 70% sequence homology thereto).

[0082] In some embodiments, the functional analog comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 89%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, or 100% homology or identity with SEQ ID NO:2. Each possibility represents a separate embodiment of the invention. In some embodiments, a homolog of the N-terminal region of a B-type lamin protein is provided that shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO:2. Each possibility represents a separate embodiment of the invention.

[0083] In some embodiments, the lamin system protein (SEQ ID NO: 1) further comprises a C-terminal region comprising the amino acid sequence: GEEERLNLTQEAPQNTSVHHVSFSSGGASAQRGVKRRRVVDVNGEDQDIDYLNRRSKLNKETVGPVGIDEVDEEGKWVRVANNSEEEQSIGGYKLVVKAGNKEASFQFSSRMKLAPHASATVWSADAGAVHHPPEVYVMKKQQWPIGDNPSARLEDSEGDTVSSITVEFSESSDPSDPADRCSIM (SEQ ID NO: 3) (including any functional analog having at least 70% sequence homology thereto).

[0084] In some embodiments, the lamin system protein (SEQ ID NO: 11) further comprises a C-terminal region comprising the amino acid sequence: NLTQEAPQNTSVHHVSFSSGGASAQRGVKRRRVVDVNGEDQDIDYLNRRSKLNKETVGPVGIDEVDEEGKWVRVANNSEEEQSIGGYKLVVKAGNKEASFQFSSRMKLAPHASATVWSADAGAVHHPPEVYVMKKQQWPIGDNPSARLEDSEGDTVSSITVEFSESSDPSDPADRCSIM (SEQ ID NO: 13) (including any functional analog having at least 70% sequence homology thereto).

[0085] In some embodiments, the lamin system protein is a B-type lamin comprising the amino acid sequence of SEQ ID NO:1, or SEQ ID NO:11, and further comprising (i) a C-terminal region (comprising the amino acid sequence of SEQ ID NO:3); (ii) an N-terminal region (comprising the amino acid sequence of SEQ ID NO:2, or SEQ ID NO:13) that comprises any sequence homologue, or functional homologue thereof; or both (i) and (ii).

[0086] In some embodiments, the functional analog comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 89%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, or 100% homology or identity with SEQ ID NO:3. Each possibility represents a separate embodiment of the present invention. In some embodiments, a homolog of the C-terminal region of a B-type lamin protein is provided that shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO:3. Each possibility represents a separate embodiment of the present invention.

[0087] In some embodiments, the lamin system protein is a B-type lamin comprising the amino acid sequence: (SEQ ID NO: 10) (including any functional analog having at least 70% sequence homology thereto).

[0088] In some embodiments, the fibers comprise A-type lamin. In some embodiments, the fibers comprise A-type lamin system proteins, and the fibers are 1 μm to 200 μm, 2 μm to 200 μm, 3 μm to 200 μm, 5 μm to 200 μm, 7 μm to 200 μm, 9 μm to 200 μm, 1 μm to 190 μm, 2 μm to 190 μm, 3 μm to 190 μm, 5 μm to 190 μm, 7 μm to 190 μm, 9 μm to 190 μm, 10 μm to 180 μm, 15 μm to 180 μm, 20 μm to 180 μm, 25 μm to 180 μm, 30 μm to 180 μm, 50 μm to 180 μm, 65 μm to 180 μm. , 10 μm to 100 μm, 15 μm to 100 μm, 20 μm to 100 μm, 25 μm to 100 μm, 30 μm to 100 μm, 50 μm to 100 μm, 65 μm to 100 μm, 10 μm to 80 μm, 15 μm to 80 μm, 20 μm to 80 μm, 25 μm to 80 μm, 30 μm to 80 μm, 50 μm to 80 μm, 65 μm to 80 μm, 10 μm to 50 μm, 15 μm to 50 μm, 20 μm to 50 μm, 25 μm to 50 μm, or 30 μm to 50 μm (including any range therebetween). Each possibility represents a separate embodiment of the present invention. In some embodiments, the fibers are characterized by a length of at least 1 cm, at least 2 cm, at least 5 cm, at least 10 cm, at least 50 cm, at least 100 cm, at least 500 cm, at least 1000 cm, or at least 10000 cm (including any value therebetween), with each possibility representing a separate embodiment of the present invention.

[0089] In some embodiments, an A-type lamin has the amino acid sequence: LQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSK(X2)AALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVE IDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLEGEEERL (SEQ ID NO: 4), including any functional analog having at least 70% sequence homology thereto. In some embodiments, X2 is Glu or Lys.

[0090] In some embodiments, an A-type lamin has the amino acid sequence: LQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSK(X2)AALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETR The repeats of the repeat region of the lamin protein include LVEIDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLE (SEQ ID NO: 7), including any functional analog having at least 70% sequence homology thereto. In some embodiments, X2 is Glu or Lys.

[0091] In some embodiments, n is an integer from 2 to 100, 3 to 100, 5 to 100, 10 to 100, 20 to 100, 30 to 100, 35 to 100, 40 to 100, 50 to 100, 2 to 80, 3 to 80, 5 to 80, 10 to 80, 20 to 80, 30 to 80, 35 to 80, 40 to 80, or 50 to 80 (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0092] In some embodiments, the functional analog comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 89%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, or 100% homology or identity with SEQ ID NO:4 or with SEQ ID NO:7. Each possibility represents a separate embodiment of the invention. In some embodiments, a homolog of the repeat region of a B-type lamin protein is provided that shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO:4 or with SEQ ID NO:7. Each possibility represents a separate embodiment of the invention.

[0093] In some embodiments, the lamin system protein further comprises an N-terminal region comprising the amino acid sequence: METPSQRRATRSGAQASSTPLSPTRITR (SEQ ID NO:5) (including any functional analog having at least 70% sequence homology thereto).

[0094] In some embodiments, the functional analog comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 89%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, or 100% homology or identity with SEQ ID NO:5. Each possibility represents a separate embodiment of the present invention. In some embodiments, a homolog of the N-terminal region of an A-type lamin protein is provided that shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO:5. Each possibility represents a separate embodiment of the present invention.

[0095] In some embodiments, the lamin system protein further comprises a C-terminal region comprising the amino acid sequence: RLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSGDNLVTRSYLLGNSSPRTQSPQNCSIM (SEQ ID NO: 6) (including any functional analog having at least 70% sequence homology thereto).

[0096] In some embodiments, the lamin system protein further comprises a C-terminal region comprising the amino acid sequence: EEERLRLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSGDNLVTRSYLLGNSSPRTQSPQNCSIM (SEQ ID NO: 8) (including any functional analog having at least 70% sequence homology thereto).

[0097] In some embodiments, the functional analog comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 89%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99%, or 100% homology or identity with SEQ ID NO:6. Each possibility represents a separate embodiment of the invention. In some embodiments, a homolog of the C-terminal region of an A-type lamin protein is provided that shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology with SEQ ID NO:6 or with SEQ ID NO:8. Each possibility represents a separate embodiment of the invention.

[0098] In some embodiments, the lamin system protein is an A-type lamin comprising the amino acid sequence of SEQ ID NO:4, or SEQ ID NO:7, and further comprising: (i) a C-terminal region (comprising the amino acid sequence of SEQ ID NO:6, or SEQ ID NO:8); (ii) an N-terminal region (comprising the amino acid sequence of SEQ ID NO:5) that comprises any sequence homologue, or functional homologue thereof; or both (i) and (ii).

[0099] In some embodiments, the Lamin system proteins of the invention comprise a repeat region comprising an amino acid sequence selected from SEQ ID NO: 1, 4, 7 or 10, or a functional analog comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 88% homology thereto (including any range therebetween), and the Lamin system proteins are oligomers comprising at least 3, at least 4, at least 5, at least 6, at least 8 polypeptide monomers. In some embodiments, the Lamin system proteins of the invention are oligomers comprising 4-8, 4-6, 6-8 or more polypeptide monomers. In some embodiments, each of the polypeptide monomers comprises an amino acid sequence selected from SEQ ID NO: 1, 4, 7 or 9, and optionally one or more of the N-terminal amino acid sequences selected from SEQ ID NO: 2 and 5, and a C-terminal amino acid sequence selected from SEQ ID NO: 6 and 8. In some embodiments, each of the polypeptide monomers comprises the same amino acid sequence. In some embodiments, at least one of the polypeptide monomers comprises a different amino acid sequence.

[0100] In some embodiments, the lamin system protein is an A-type lamin comprising the amino acid sequence of (SEQ ID NO:9) (including any functional analog having at least 70% sequence homology thereto).

[0101] In some embodiments, the fibers comprising A-type lamins described above further comprise a B-type lamin.

[0102] In some embodiments, the fibers of the invention comprise or consist essentially of lamin system proteins characterized by an α-helix:β-sheet ratio of 5:1 to 1:1, 4.55:1 to 1:1, 4:1 to 1:1, 3:1 to 1:1, 2:1 to 1:1, 5:1 to 3:1, 4.55:1 to 3:1, 4:1 to 3:1, 5:1 to 2:1, 4.55:1 to 2:1, 4:1 to 2:1, or 3:1 to 2:1 (including any range therebetween), where the α-helix:β-sheet ratio is determined by Raman. Each possibility represents a separate embodiment of the invention. An exemplary Raman-based determination of α-helix:β-sheet ratio is described in Example 2. Additional methods include FTIR and X-ray scattering.

[0103] In some embodiments, the fibers are elongated fibers. In some embodiments, the elongated fibers are characterized by a relative content of α-helical lamin system proteins in the range of about 30 to about 99%, about 30 to about 45%, about 30 to about 40% (including any range therebetween). In some embodiments, the elongated fibers are characterized by a relative content of β-sheet lamin system proteins in the range of about 30 to about 50%, about 30 to about 45%, about 30 to about 40%, about 30 to about 80%, about 30 to about 60%, about 60 to about 99%, about 40 to about 80%, about 40 to about 90% (including any range therebetween). The relative content of α-helical or β-sheet lamin system proteins is determined by Raman as described herein below, relative to the total lamin system protein content of the fiber.

[0104] As used herein, the term "stretched fiber" refers to a fiber or filament that has undergone a stretching or drawing process. The stretching or drawing process refers to a process of pulling a long fiber or filament into alignment along its longitudinal axis. The drawing is typically performed to a strain less than the break strain of the fiber (e.g., in the range of 100-400%, or 100-1000%), as disclosed below.

[0105] In some embodiments, the fibers of the invention comprise or consist essentially of a lamin system protein characterized by an alpha-helix to beta-sheet transition upon stretching the fiber in the elastic region. In some embodiments, the fibers of the invention comprise or consist essentially of a lamin system protein characterized by an alpha-helix to beta-sheet transition upon stretching the fiber to about 6% strain. In some embodiments, the alpha-helix to beta-sheet transition comprises at least 10%, at least 20%, at least 30%, 5-50%, 10-50%, 20-50%, 20-40%, 10-70%, 10-60% transition (including any range therebetween).

[0106] In some embodiments, a fiber of the invention comprises, or consists essentially of, a lamin system protein characterized by about 20% to about 90%, about 20% to about 60%, 20% to 50%, 20% to 90%, 3% to 50%, 4% to 50%, 5% to 50%, 7% to 50%, 10% to 50%, 15% to 50%, 20% to 50%, 30% to 50%, 3% to 40%, 4% to 40%, 5% to 40%, 7% to 40%, 10% to 40%, 15% to 40%, 20% to 40%, 30% to 40%, 3% to 25%, 4% to 25%, 5% to 25%, 7% to 25%, 10% to 25%, or 15% to 25% alpha helix to beta sheet transition (including any range therebetween). Each possibility represents a separate embodiment of the invention. The alpha helix to beta sheet transition is determined by Raman after stretching the fiber. The alpha helix to beta sheet transition refers to the change in the alpha helix content of the fiber relative to the initial alpha helix content before stretching. In some embodiments, the fibers of the invention undergo an alpha helix to beta sheet transition in the elastic region. In some embodiments, the fibers of the invention undergo an alpha helix to beta sheet transition (e.g., at least 10%, or at least 20% transition) when stretched to about 6%, about 10% or more strain, e.g., less than the breaking strain, 5-1000%, about 5-100%, about 5-80% strain (including any range therebetween).

[0107] In some embodiments, the elongated fibers of the invention comprise or consist essentially of lamin system proteins characterized by an alpha helix to beta sheet ratio of about 2:1 to about 1:1 (including any range therebetween), and the elongated fibers are characterized by a strain of about 6 to about 100%, about 6 to about 80%, about 6 to about 50%, about 6 to about 60%, about 6 to about 70%, about 6 to about 90% (including any range therebetween).

[0108] In some embodiments, each repeat independently has a molecular weight in the range of 20 kDa to 80 kDa, 20 kDa to 70 kDa, 20 kDa to 60 kDa, 20 kDa to 55 kDa, or 20 kDa to 50 kDa (including any range therebetween), with each possibility representing a separate embodiment of the present invention.

[0109] In some embodiments, the fibers include a plurality of lamin system proteins arranged in the form of paracrystals, in some embodiments, each paracrystal is characterized by a dimension selected from: (i) a width between 1 nm and 500 nm; (ii) a length between 0.5 mm and 1 cm, or both (i) and (ii).

[0110] In some embodiments, each quasicrystal is characterized by a width of 1 nm to 500 nm, 2 nm to 500 nm, 5 nm to 500 nm, 15 nm to 500 nm, 50 nm to 500 nm, 100 nm to 500 nm, 250 nm to 500 nm, 1 nm to 300 nm, 2 nm to 300 nm, 5 nm to 300 nm, 15 nm to 300 nm, 50 nm to 300 nm, 100 nm to 300 nm, 1 nm to 100 nm, 2 nm to 100 nm, 5 nm to 100 nm, 15 nm to 100 nm, or 50 nm to 100 nm (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0111] In some embodiments, each quasicrystal is 0.5 mm to 1 cm, 0.7 mm to 1 cm, 0.9 mm to 1 cm, 1 mm to 1 cm, 10 mm to 1 cm, 30 mm to 1 cm, 50 mm to 1 cm, 100 mm to 1 cm, 500 mm to 1 cm, 700 mm to 1 cm, 0.5 mm to 900 mm, 0.7 mm to 900 mm, 0.9 mm to 900 mm, 1 mm to 900 mm, 10 mm to 900 mm, 30 mm to 9 In some embodiments, the length may be characterized as 00 mm, 50 mm to 900 mm, 100 mm to 900 mm, 500 mm to 900 mm, 700 mm to 900 mm, 0.5 mm to 500 mm, 0.7 mm to 500 mm, 0.9 mm to 500 mm, 1 mm to 500 mm, 10 mm to 500 mm, 30 mm to 500 mm, 50 mm to 500 mm, or 100 mm to 500 mm (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0112] As used herein, the term "quasicrystalline" refers to nanofilaments (obtained upon association of tetrameric protofilaments), which are unidirectional nanofilaments aligned along the longitudinal axis of the fiber. As used herein, "protofilament" refers to the structure formed by polymerization of lamin dimers. In some embodiments, the dimeric polymers associate to form tetrameric protofilaments.

[0113] In some embodiments, the lamin system proteins are assembled in the fibers of the invention in the form of lamin dimers. In some embodiments, the lamin system proteins are assembled in the fibers of the invention in the form of protofilaments. In some embodiments, the protofilaments are essentially unidirectionally aligned in the fiber. In some embodiments, the protofilaments are aligned along the longitudinal axis of the fiber. In some embodiments, each protofilament comprises multiple lamin dimers (e.g., a polymer comprising multiple lamin dimers). In some embodiments, the lamin system proteins are assembled in the fibers of the invention in the form of tetrameric protofilaments. In some embodiments, the lamin system proteins are assembled in the fibers of the invention in the form of paracrystalline (or nanofilaments) comprising tetrameric protofilaments.

[0114] In some embodiments, the lamin system protein is an isolated protein.

[0115] As used herein, the term "isolated protein" refers to a protein that is essentially free from contaminating cellular components, such as carbohydrates, lipids, or other proteinaceous impurities that naturally accompany the nucleic acid. Typically, an isolated protein preparation contains a highly purified form of the protein, such as at least about 80% pure, at least about 90% pure, at least about 95% pure, more than 95% pure, or more than 99% pure. In some embodiments, the isolated protein is a synthetic protein. Protein synthesis is well known in the art and can be performed, for example, by heterologous expression in transformed cells as exemplified herein.

[0116] In some embodiments, the lamin system protein is a recombinant protein. In some embodiments, the fiber is obtained by expression in a recombinant cell. In some embodiments, the fiber is obtained by expression in a bacterium. In some embodiments, the bacterium is Escherichia coli.

[0117]

[0118]

[0119] In some embodiments, the artificial vector comprises a plasmid. In some embodiments, the artificial vector comprises an Agrobacterium comprising an artificial nucleic acid molecule or is an Agrobacterium comprising an artificial nucleic acid molecule. In some embodiments, the artificial vector is an expression vector. In some embodiments, the artificial vector is a plant expression vector. In some embodiments, the artificial vector is for use in the expression of an AAE-encoding nucleic acid sequence disclosed herein. In some embodiments, the artificial vector is for use in the heterologous expression of an AAE-encoding nucleic acid sequence disclosed herein in a cell, tissue, or organism. In some embodiments, the artificial vector is for use in the production or production of acyl-coenzyme A (acyl-CoA) in a cell, tissue, or organism.

[0120] The expression of polynucleotides in cells is well known to those skilled in the art. It can be carried out by transfection, viral infection, or direct alteration of the genome of the cell, among many other methods. In some embodiments, the polynucleotide is in an expression vector, such as a plasmid or a viral vector. The vector nucleic acid sequence generally contains at least one origin of replication for propagation in the cell, and optionally additional elements, such as heterologous polynucleotide sequences, expression control elements (e.g., promoters, enhancers), selectable markers (e.g., antibiotic resistance), poly-adenine sequences.

[0121] The vector can be a DNA plasmid delivered via non-viral or viral methods. The viral vector can be a retroviral vector, a herpes virus vector, an adenovirus vector, an adeno-associated virus vector, a veraliprid virus vector, or a pox virus vector. Barley stripe mosaic virus (BSMV), tobacco rattle virus, and cabbage leaf curl virus (CbLCV) can also be used. The promoter can be active in plant cells. The promoter can be a viral promoter.

[0122] In some embodiments, the polynucleotide as disclosed herein is operably linked to a promoter. The term "operably linked" is intended to mean that the nucleotide sequence of interest is linked to one or more regulatory elements or elements in a manner that allows expression of the nucleotide sequence (e.g., in an in vitro transcription / translation system or in a host cell when a vector is introduced into a host cell). In some embodiments, a promoter is operably linked to the polynucleotide of the present invention. In some embodiments, the promoter is a heterologous promoter. In some embodiments, the promoter is an endogenous promoter.

[0123] In some embodiments, vectors are introduced into cells by standard methods including electroporation (e.g., as described in From et al., Proc. Natl. Acad. Sci. USA 82, 5824 (1985)), heat shock, infection with viral vectors, high velocity ballistic penetration by small particles with nucleic acid either within the matrix of small beads or particles or on a surface (Klein et al., Nature 327.70-73 (1987)), e.g. coated particles, and needle-shaped particles, biolistic use of Agrobacterium Ti plasmids, etc.

[0124] The term "promoter" as used herein refers to a group of transcriptional control modules clustered around the initiation site of RNA polymerase, i.e., RNA polymerase II. Promoters are composed of separate functional modules, each consisting of approximately 7-20 bp of DNA and containing one or more recognition sites for transcriptional activator or repressor proteins. Promoters may extend upstream or downstream of the transcription start site and may be any size ranging from a few base pairs to several kilobases.

[0125] In some embodiments, the polynucleotide is transcribed by RNA polymerase II (RNAP II and Pol II). RNAP II is an enzyme found in eukaryotic cells that is known to catalyze the transcription of DNA to synthesize the precursors of mRNA and most snRNAs and microRNAs.

[0126] In some embodiments, recombinant viral vectors that provide advantages such as systemic infection and target specificity are used for in vivo expression. In one embodiment, systemic infection is inherent in, for example, the life cycle of retroviruses, a process in which a single infected cell produces many progeny virions that infect neighboring cells. In one embodiment, this results in rapid infection of a large area, the majority of which were not initially infected by the original viral particle. In one embodiment, a viral vector that cannot spread systemically is produced. In one embodiment, this feature can be useful when the desired purpose is to introduce a specific gene only into a local number of target cells.

[0127] In some embodiments, a plant viral vector is used. In some embodiments, a wild type virus is used. In some embodiments, a degraded virus is used as known in the art. In some embodiments, Agrobacterium is used to introduce the vector of the invention into the virus.

[0128] In some embodiments, expression vectors containing regulatory elements from eukaryotic viruses, such as retroviruses, are used by the present invention. SV40 vectors include pSVT7 and pMT2. In some embodiments, vectors derived from bovine papilloma virus include pBV-1MTHA, and vectors derived from Epstein-Barr virus include pHEBO, and p2O5. Other exemplary vectors include pMSG, pAV009 / A+, pMTO10 / A+, pMAMneo-5, baculovirus pDSVE, and any other vector that allows expression of a protein under the direction of the SV-40 early promoter, SV-40 late promoter, metallothionein promoter, mouse mammary tumor virus promoter, Rous sarcoma virus promoter, polyhedrin promoter, or other promoters shown to be effective for expression in eukaryotic cells.

[0129] A variety of methods can be used to introduce the expression vector of the present invention into cells. Such methods are generally described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Springs Harbor Laboratory, New York (1989, 1992), Ausubel et al., Current Protocols in Molecular Biology, John Wiley and Sons, Baltimore, Md. (1989), Chang et al., Somatic Gene Therapy, CRC Press, Ann Arbor, Mich. (1995), Vega et al., Gene Targeting, CRC Press, Ann Arbor Mich. (1995), Vectors: A Survey of Molecular Cloning Vectors and Their Uses, Butterworths, Boston Mass. (1988) and Gilboa et al. [Biotechniques 4(6):504-512, 1986] and include, for example, stable or transient transfection, lipofection, electroporation and infection with recombinant viral vectors. See also US Pat. Nos. 5,464,764 and 5,487,992 for positive-negative selection methods. In one embodiment, a plant expression vector is used. In one embodiment, the expression of the polypeptide coding sequence is driven by some promoters. In some embodiments, viral promoters are used, such as the 35S RNA and 19S RNA promoters of CaMV [Brisson et al., Nature 310:511-514 (1984)], or the coat protein promoter for TMV [Takamatsu et al., EMBO J.6:307-311 (1987)]. In another embodiment, a plant promoter is used, such as the small subunit of RUBISCO [Coruzzi et al., EMBO J.3:1671-1680 (1984); and Brogli et al., Science 224:838-843 (1984)] or a heat shock promoter, such as soybean hsp17.5-E or hsp17.3-B [Gurley et al., Mol. Cell. Biol.6:559-565 (1986)]. In one embodiment, constructs are introduced into plant cells using Ti plasmid, Ri plasmid, plant virus vector, direct DNA transformation, microinjection, electroporation and other techniques well known to those skilled in the art.See, for example, Weissbach&Weissbach [Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp421-463(1988)].Other expression systems, such as insect and mammalian host cell systems well known in the art, can also be used according to the present invention.

[0130] It will be understood that, in addition to containing the necessary elements for the transcription and translation of the inserted coding sequence (encoding a polypeptide), the expression constructs of the invention can also contain sequences engineered to optimize the stability, production, purification, yield, or activity of the expressed polypeptide.

[0131] In some embodiments, the artificial vector comprises a polynucleotide encoding a protein comprising an amino acid sequence described herein.

[0132] In some embodiments, the lamin system protein is an isolated protein.

[0133] As used herein, the term "isolated protein" refers to a protein that is essentially free from contaminating cellular components, such as carbohydrates, lipids, or other proteinaceous impurities that naturally accompany the nucleic acid. Typically, an isolated protein preparation contains a highly purified form of the protein, such as at least about 80% pure, at least about 90% pure, at least about 95% pure, more than 95% pure, or more than 99% pure. In some embodiments, the isolated protein is a synthetic protein. Protein synthesis is well known in the art and can be performed, for example, by heterologous expression in transformed cells as exemplified herein.

[0134] In some embodiments, the fibers of the invention further comprise a coating. In some embodiments, Lamin system protein fibers comprising a coating are referred to herein as "coated fibers." In some embodiments, the fibers of the invention are coated fibers.

[0135] In some embodiments, the coated fibers comprise Lamin system protein fibers of the invention in contact with a coating. In some embodiments, the surface of the fiber is in contact with the coating. In some embodiments, the exterior surface of the Lamin system protein fiber is in contact with the coating. In some embodiments, at least a portion of the Lamin system protein fiber surface (i.e., at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97%, at least 99%, at least 99.9%, at most 90%, at most 100%, at most 95%, at most 97%) is in contact with the coating.

[0136] In some embodiments, the term "contacting" encompasses a lamin system protein fiber (e.g., an outer portion of the fiber) being bound to the coating via a non-covalent bond (e.g., including a physical bond or interaction). In some embodiments, the bond is via physical adsorption. In some embodiments, the coating is adsorbed to the lamin system protein fiber. In some embodiments, the coating is embedded into at least a portion of the lamin system protein fiber. In some embodiments, the coated fiber comprises a lamin system protein fiber of the invention stably bound to a coating.

[0137] As used herein, the term "stably bonded" refers to the ability of the coated fiber to substantially maintain its structural, physical and / or chemical properties when stored under appropriate conditions for periods ranging from up to 1 month, up to 1 year, up to 3 years, or up to 10 years or more (including any ranges therebetween). In some embodiments, the fibers of the present invention are called stable if they substantially maintain their structure (e.g., shape, and / or dimensions, e.g., thickness, length, etc.), in some embodiments, substantially as described herein. In some embodiments, appropriate conditions include exposure to ambient atmosphere, UV / Vis light irradiation, and / or temperatures of -50 to 70°C, -50 to 60°C, -50 to 50°C, -50 to 0°C, 0 to 10°C, 10 to 30°C, 30 to 50°C, 50 to 70°C, 70 to 100°C, and any ranges therebetween.

[0138] In some embodiments, a coated fiber is referred to as stable when the coated fiber is substantially free of crumbling, cracking, deformation, or any other surface irregularities or defects throughout its composition or physical structure.

[0139] In some embodiments, the coating is in the form of a coating layer. In some embodiments, the terms "coating" and "coating layer" are used interchangeably herein.

[0140] In some embodiments, the coating is in the form of a film. In some embodiments, the coating forms a substantially uniform layer over the Lamin system protein fibers. In some embodiments, the coating layer is a uniform layer.

[0141] "Uniform" or "homogeneous" when referring to a layer or film is meant to refer to a size (or thickness) distribution that varies, for example, within ±50%, ±40%, ±30%, ±20%, ±10%, ±5%, or less (including any value therebetween).

[0142] In some embodiments, the term "layer" refers to a substantially uniform thickness of a substantially uniform material. In some embodiments, a layer or film comprises a single layer or multiple layers. In some embodiments, the terms layer and film are used interchangeably herein.

[0143] In some embodiments, the coating (or coating layer) is at least 1 nm, at least 10 nm, at least 5 nm, at least 50 nm, at least 100 nm, at least 500 nm, at least 1 μm, at least 5 μm, at least 10 μm, at least 50 μm (including any range therebetween). In some embodiments, the coating (or coating layer) is characterized by a thickness of about 1 nm to about 50 μm, about 1 nm to about 50 μm, about 1 nm to about 30 μm, about 1 nm to about 20 μm, about 1 nm to about 10 μm, about 1 nm to about 1 μm, about 1 nm to about 0.1 μm, about 10 nm to about 50 μm, about 10 nm to about 5 μm, about 10 nm to about 1 μm, about 10 nm to about 0.1 μm, about 1 nm to about 500 nm, about 1 nm to about 100 nm, about 10 to about 1000 μm, about 100 nm to about 10 μm, about 10 to about 50 μm (including any range therebetween). The terms "thickness" and "thickness" refer to an average value.

[0144] In some embodiments, the coating comprises a film-forming agent. In some embodiments, the coating is a hydrophobic coating. In some embodiments, the hydrophobic coating comprises or consists essentially of a water-immiscible compound. In some embodiments, the water-immiscible compound is configured to form a stable film on the lamin system protein fibers. In some embodiments, the water-immiscible compound is a water-immiscible small molecule, or a water-immiscible polymer.

[0145] In some embodiments, the coating comprises vegetable oils, mineral oils, fatty acids, isobutyl stearate, tallow fatty acid 2-ethylhexyl esters, polyol carboxylic acid esters, coconut fatty acid esters of glycerol, alkoxylated glycerol, silicones, dimethylpolysiloxanes, polyalkylene glycols, polyethylene oxides, and propylene oxide copolymers (including any salts, any combinations, and any copolymers thereof).

[0146] In some embodiments, the hydrophobic coating is selected from oils, fats, fatty acids, fatty acid esters, fatty alcohols, glycerides (e.g., mono-, di-, and / or tri-glycerides), phospholipids, lipids, solid lipids, vegetable oils, essential oils, vegetable oils, isobutyl stearate, tallow fatty acid 2-ethylhexyl esters, polyol carboxylic acid esters, glycerides, coconut fatty acid esters of glycerol, alkoxylated glycerol, silicone oils, mineral oils, dimethylpolysiloxanes, polysiloxanes, polysilanes, and waxes (including any salts, any combinations, and any copolymers thereof).

[0147] According to one aspect of some embodiments of the present invention, the fiber has the following properties: yield strength of 1 MPa to 1000 MPa; tensile strength of -1 MPa to 1000 MPa; breaking strain of 10% to 500%; and 30 MJ / m 3 ~1000MJ / m 3and a Young's modulus between 0.001 GPa and 30 GPa.

[0148] In some embodiments, the fibers of the present invention have a compressibility of 1 MPa to 1000 MPa, 5 MPa to 1000 MPa, 10 MPa to 1000 MPa, 20 MPa to 1000 MPa, 30 MPa to 1000 MPa, 50 MPa to 1000 MPa, 70 MPa to 1000 MPa, 100 MPa to 1000 MPa, 250 MPa to 1000 MPa, 300 MPa to 1000 MPa, 500 MPa to 1000 MPa, 1 MPa to 700 MPa, 5 MPa to 700 MPa, 10 MPa to 700 MPa, 20 MPa to 700 MPa, 300 MPa to 700 MPa, The material may be characterized by a yield strength of 100 MPa to 700 MPa, 50 MPa to 700 MPa, 70 MPa to 700 MPa, 100 MPa to 700 MPa, 250 MPa to 700 MPa, 300 MPa to 700 MPa, 500 MPa to 700 MPa, 1 MPa to 300 MPa, 5 MPa to 300 MPa, 10 MPa to 300 MPa, 20 MPa to 300 MPa, 30 MPa to 300 MPa, 50 MPa to 300 MPa, 70 MPa to 300 MPa, or 100 MPa to 300 MPa (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0149] In some embodiments, the fibers of the present invention have a compressibility of 1 MPa to 1000 MPa, 5 MPa to 1000 MPa, 10 MPa to 1000 MPa, 20 MPa to 1000 MPa, 30 MPa to 1000 MPa, 50 MPa to 1000 MPa, 70 MPa to 1000 MPa, 100 MPa to 1000 MPa, 250 MPa to 1000 MPa, 300 MPa to 1000 MPa, 500 MPa to 1000 MPa, 1 MPa to 700 MPa, 5 MPa to 700 MPa, 10 MPa to 700 MPa, 20 MPa to 700 MPa, 300 MPa to 700 MPa, The tensile strength may be characterized by a range of from 100MPa to 700MPa, 50MPa to 700MPa, 70MPa to 700MPa, 100MPa to 700MPa, 250MPa to 700MPa, 300MPa to 700MPa, 500MPa to 700MPa, 1MPa to 300MPa, 5MPa to 300MPa, 10MPa to 300MPa, 20MPa to 300MPa, 30MPa to 300MPa, 50MPa to 300MPa, 70MPa to 300MPa, or 100MPa to 300MPa (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0150] In some embodiments, the fibers of the present invention are characterized by a breaking strain of 10%-500%, 20%-500%, 50%-500%, 90%-500%, 100%-500%, 250%-500%, 10%-350%, 20%-350%, 50%-350%, 90%-350%, 100%-350%, 250%-350%, 50-1000%, about 100-about 1000%, about 100-about 800%, about 100-about 400%, about 100-about 500%, about 100-about 600%, about 100-about 700%, about 100-about 900% (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0151] In some embodiments, the fibers of the present invention have a thermal conductivity of 30 MJ / m 3 ~1000MJ / m 3 , 50MJ / m 3 ~1000MJ / m 3 , 70MJ / m 3 ~1000MJ / m 3 , 100MJ / m 3~1000MJ / m 3 , 300MJ / m 3 ~1000MJ / m 3 , 500MJ / m 3 ~1000MJ / m 3 , 700MJ / m 3 ~1000MJ / m 3 , 30MJ / m 3 ~700MJ / m 3 , 50MJ / m 3 ~700MJ / m 3 , 70MJ / m 3 ~700MJ / m 3 , 100MJ / m 3 ~700MJ / m 3 , 300MJ / m 3 ~700MJ / m 3 , 500MJ / m 3 ~700MJ / m 3 , 30MJ / m 3 ~500MJ / m 3 , 50MJ / m 3 ~500MJ / m 3 , 70MJ / m 3 ~500MJ / m 3 , 100MJ / m 3 ~500MJ / m 3 , or 300 MJ / m 3 ~500MJ / m 3 (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0152] In some embodiments, the fibers of the present invention have a viscosity of 0.001 GPa to 30 GPa, 0.005 GPa to 30 GPa, 0.009 GPa to 30 GPa, 0.01 GPa to 30 GPa, 0.05 GPa to 30 GPa, 0.1 GPa to 30 GPa, 1 GPa to 30 GPa, 10 GPa to 30 GPa, 0.001 GPa to 20 GPa, 0.005 GPa to 20 GPa, 0.009 GPa to 20 GPa, 0.01 GPa to 20 GPa The physical properties described above are characterized by a Young's modulus of 0.05 GPa to 20 GPa, 0.1 GPa to 20 GPa, 1 GPa to 20 GPa, 10 GPa to 20 GPa, 0.001 GPa to 10 GPa, 0.005 GPa to 10 GPa, 0.009 GPa to 10 GPa, 0.01 GPa to 10 GPa, 0.05 GPa to 10 GPa, 0.1 GPa to 10 GPa, or 1 GPa to 10 GPa (including any range therebetween). Each possibility represents a separate embodiment of the present invention. The physical properties described above encompass any of the fibers disclosed herein, including lamin-protein based fibers, and coated fibers.

[0153] According to another aspect of some embodiments of the present invention, there is provided a fiber comprising a B-type lamin system protein, the fiber being characterized by (i) a diameter of between 10 μm and 180 μm; (ii) the B-type lamin system protein comprises an amino acid set forth in SEQ ID NO: 1, or SEQ ID NO: 11 (including any functional analog having at least 70% sequence homology thereto), and when X1 is Gln, the fiber has (i) a residual amount of alcohol; (ii) a residual amount of alcohol of 190 MJ / m 3 and (ii) the protein is characterized by an α-helix to β-sheet transition upon elongation of 3% to 50%. In some embodiments, the fiber further comprises either: i. an N-terminal region comprising the amino acid sequence SEQ ID NO:2 or any functional analog having at least 70% sequence identity thereto; ii. a C-terminal region comprising the amino acid sequence SEQ ID NO:3 or any functional analog having at least 70% sequence identity thereto.

[0154] In some embodiments, the fibers of the present invention have a thermal conductivity of 195 MJ / m 3 Super, 200MJ / m 3 Super, 205MJ / m 3 Super, 250MJ / m 3 Super, 270MJ / m 3 Super, 290MJ / m 3 Super, 300MJ / m 3 Super, 500MJ / m 3 or over 700MJ / m 3 Each possibility characterized by a toughness of greater than (including any value therebetween) represents a separate embodiment of the present invention.

[0155] In some embodiments, the fibers are for use in biomaterials and biomimetic materials.

[0156] In some embodiments, the fibers are for use in the manufacture of scaffolds suitable for the field of regenerative medicine, for example, tissue engineering and tissue graft production.

[0157] In some embodiments, the fibers described above are for use in the manufacture of wound closure or coverage systems, such as the manufacture of suture materials and wound dressings.

[0158] In some embodiments, the fibers are for use in medical devices such as medical adhesive strips, skin grafts, ligament substitutes, surgical meshes, membranes, and filters.

[0159] In some embodiments, the fibers are for use in cosmetics and drug delivery.

[0160] In some embodiments, the fibers are for use in a wide range of industrial and commercial products, such as films (e.g., transparent films), fabrics for apparel, bulletproof vest linings, fabrics for containers, bag or purse straps, cables, ropes, adhesive bonding materials, non-adhesive bonding materials, strapping materials, automobile covers and parts, aircraft structural materials, weather resistant materials, flexible bulkhead materials, and sporting goods.

[0161] In some embodiments, the fibers are for use in coatings, such as coatings for textile and leather products, thereby imparting stability and durability to the coated product or imparting additional properties to the coated product, such as water repellency.

[0162] In some embodiments, the fibers described above can be combined with other materials to produce a product or article. In some embodiments, the fibers described above can be combined with other materials to obtain a composite. In some embodiments, the composite is stable, the term "stable" being as described herein. In some embodiments, a composite is provided that includes the fibers of the present invention and an additional polymer. In some embodiments, the composite further includes additives such as binders, crosslinkers, plasticizers, stabilizers, fillers, etc. In some embodiments, the polymer content of the composite consists essentially of the fibers of the present invention and the additional polymer. In some embodiments, 50-99%, 50-95%, 50-90%, 50-80%, 50-70% of the polymer content of the composite consists of the fibers of the present invention and the additional polymer. In some embodiments, the additional polymer forms the matrix of the composite, and the fibers of the present invention provide mechanical reinforcement to the matrix. In some embodiments, the additional polymer is compatible with the fibers of the present invention to obtain a stable composite.

[0163] In some embodiments, the composite comprises 1-50%, 1-30%, 1-10%, 1-20%, 10-50%, 20-50%, 30-50%, 10-60%, 10-70% w / w of the fibers of the invention and further comprises an additional polymer. In some embodiments, the additional polymer is transparent (e.g., 70-100% light transmission). In some embodiments, the additional polymer comprises a synthetic polymer, including a thermoplastic polymer or a thermosetting polymer. In some embodiments, the synthetic polymer is a non-biodegradable polymer. In some embodiments, the synthetic polymer is a biodegradable polymer. In some embodiments, the synthetic polymer is a crosslinked polymer (also referred to herein as a "cured polymer").

[0164] In some embodiments, the additional polymer comprises a cured resin. Examples of suitable resins include, but are not limited to, epoxy resins, unsaturated polyester resins, vinyl ester resins, methacrylate resins (including, for example, acrylates and esterified acrylates or other acrylate-based resins), fluorocarbon resins, and phenolic resins, or any combination thereof.

[0165] In some embodiments, the additional polymer is a polyolefin (e.g., polyethylene, polypropylene), polyester, polystyrene, C1-C8 alkylstyrene, polyvinyl chloride, polycarbonate, polyamide (e.g., nylon, etc.), polyurethane, aromatic polyetherketone resin, polyphenylene sulfide, acrylonitrile butadiene styrene (ABS), styrene acrylonitrile copolymer (SAN); poly(vinylcyclohexane); PMMA / poly(vinyl fluoride) blends; poly(phenylene oxide) alloys; styrenic block copolymers; polyimides; polysulfones; poly(vinyl chloride). ); poly(dimethylsiloxane) (PDMS); polyurethanes; unsaturated polyesters; poly(alkane terephthalates) such as poly(ethylene terephthalate) (PET); poly(alkane naphthalates) such as poly(ethylene naphthalate) (PEN); ionomers; vinyl acetate / polyethylene copolymers; cellulose acetate; cellulose acetate butyrate; fluoropolymers; poly(styrene)-poly(ethylene) copolymers; poly(carbonate) / aliphatic PET blends and PET and PEN copolymers (including polyolefin-based PET and PEN), including any copolymers and any mixtures thereof.

[0166] In some embodiments, the synthetic biodegradable polymer is or includes any one of polyglycolic acid, polyorthoesters, polyphosphoesters, polyanhydrides, polyesteramides, polyamino acids (e.g., random polyamino acids), polyimines, poly(L-lactic acid), poly(caprolactone), poly(lactic acid-coglycolic acid), poly(3-hydroxybutyric acid), poly(sebacic acid), poly(adipic acid), polyphosphazene, poly(dioxanone), poly-β-hydroxybutyrate-co-β-hydroxyvalerate (PHBV), and PBAT (including any copolymers thereof and any mixtures thereof).

[0167] In some embodiments, the additional polymer comprises a polymer derived from a natural product. In some embodiments, the additional polymer comprises a biodegradable polymer derived from a natural product. In some embodiments, the additional polymer comprises cellulose, silk, keratin and collagen, or any mixture or copolymer thereof. The composite material can be utilized in the manufacture of paper or skin and hair care products, so that the paper or skin and hair care products have improved properties, such as improved tensile strength or tear strength.

[0168] Goods According to some embodiments, the present invention provides articles comprising the fibers described herein, including inter alia coated fibers. In some embodiments, the articles comprise fibers comprising a lamin system protein, the fibers being characterized by a diameter between 10 μm and 180 μm. In some embodiments, the lamin system protein comprises an A-type lamin, a B-type lamin, or both, as described above.

[0169] In some embodiments, the article is in the form of a yarn comprising a plurality of the fibers of the present invention. In some embodiments, the article is in the form of a woven or nonwoven substrate. In some embodiments, the article is in the form of a filament, film, foam, thread, sphere, particle, microcapsule, hydrogel, or nanofibril. In some embodiments, the fibers of the present invention can be used or incorporated into any article where desired properties are, for example, high toughness and stiffness.

[0170] Non-limiting examples of articles include sutures, surgical meshes, medical adhesive strips, meshes, skin grafts, fat grafts, cosmetics, dermal fillers ligament substitutes, apparel fabrics for drug eluting / delivery devices, bulletproof vest linings, cables, tubes, films, ropes, fishing lines, tires, gloves, catheters, hoses, shoe soles, sporting goods, and reinforced composite forms.

[0171] In some embodiments, the article is characterized by at least one improved mechanical property compared to the properties of an article not containing the fibers, the property being selected from the group consisting of Young's modulus, tensile strength, strain at break, yield point, toughness, work to failure, impact strength, tear strength, flexural modulus, flexural strain, and stress at a particular elongation.

[0172] mechanical properties In some embodiments, the disclosed articles are characterized by improved mechanical properties compared to a reference article. In some embodiments, the term "reference article" refers to the same article without the fibers disclosed herein. In some embodiments, the term "reference article" refers to self-assembled fibers. In some embodiments, the term "reference article" refers to fibers processed from an aqueous coagulation bath. In some embodiments, the term "reference article" refers to uncoated fibers of the invention (i.e., lamin system protein fibers).

[0173] "Improved mechanical properties" means having more desirable mechanical properties.

[0174] In some embodiments, the improved mechanical properties refer to elastic modulus. In some embodiments, the phrase "elastic modulus" refers to Young's modulus. In some embodiments, the phrase "elastic modulus" is determined by the response of a material to the application of a tensile stress (e.g., by procedures known in the art).

[0175] In some embodiments, the improved mechanical properties refer to flexural modulus.As used herein and in the art, flexural modulus "flexural modulus" is the ratio of stress to strain in bending deformation, or the tendency of a material to bend.Flexural modulus can be determined from the slope of the stress-strain curve.

[0176] In some embodiments, the properties are selected from, but not limited to, Young's modulus, tensile strength, strain at break, yield point, toughness, abrasion resistance, stiffness, creep resistance, work of fracture, stress and elongation. In some embodiments, tests such as abrasion tests can also be performed according to DIN.

[0177] Stiffness refers to the slope of the linear portion of the load-deformation curve. Work of failure refers to the area under the load-deformation curve before failure. Each of these can be measured and calculated by methods standard known in the art. In some embodiments, the terms "stiffness" and "Young's modulus" are used interchangeably herein.

[0178] In some embodiments, the tensile strength of a material refers to the maximum amount of tensile stress that the material can be subjected to before it fails, eg, breaks.

[0179] In some embodiments, the term "tensile strength" as used herein is the maximum amount of force, measured for example in Newtons, that a material can withstand without or before tearing, breaking, or necking forming microcracks or cracks.

[0180] "Tearing, breaking, necking to form microcracks or cracks" is meant to refer to permanent deformation. In some embodiments, the term "permanent deformation" does not include microcracks or cracks. In some embodiments, "permanent deformation" is meant to refer to at least 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, or 1% (including any value therebetween) of an original dimension or structure.

[0181] In some embodiments, the term "strain at break" refers to the strain (displacement) at break as determined by a stress-strain curve, for example in a tensile test.

[0182] In some embodiments, the term "yield point" refers to the stress at which the stress-strain curve has a plateau and reaches the elastic limit.

[0183] As used herein, "creep" is a measure of the change in tensile strain when a polymer sample is subjected to a constant tensile stress, such as gravity or applied mechanical or physical stress. In other words, creep is the tendency of a solid material to move slowly or permanently deform under the influence of a constant tensile stress. As used herein, the term "creep resistance" refers to the ability of a polymer to resist any kind of strain when subjected to a load over an extended period of time. "Improved creep resistance" can refer to an improvement of, for example, 20% of the time for a tensile strain of, for example, 5%.

[0184] In some embodiments, the term "stress at extension" refers to the force acting on a material in an elongated state. For example, "stress at 100% extension" refers to the force acting on a material that has been stretched to twice its length.

[0185] In some embodiments, one or more properties selected from Young's modulus, tensile strength, yield point, and stress at extension are improved, for example, by at least 1%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 50%, at least 100%, at least 200%, or at least 500%.

[0186] In some embodiments, one or more properties selected from Young's modulus, tensile strength, yield point, and stress at extension are improved, for example, by at least 100%, at least 150%, at least 250%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, at least 600%, at least 650%, at least 700%, at least 750%, at least 800%, at least 850%, at least 900%, at least 1000%, at least 1500%, at least 2000%, at least 2500%, or at least 3000%.

[0187] In some embodiments, at least two properties selected from Young's modulus, tensile strength, yield point, and stress at extension are improved, for example, by at least 1%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 50%, at least 100%, at least 200%, or at least 500%.

[0188] In some embodiments, at least three properties selected from Young's modulus, tensile strength, yield point, and stress at extension are improved, for example, by at least 1%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 50%, at least 100%, at least 200%, or at least 500%.

[0189] In some embodiments, the Young's modulus is increased by, for example, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 50%, at least 100%, at least 200%, or at least 500%.

[0190] In some embodiments, the tensile strength is improved by, for example, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, or at least 50%.

[0191] In some embodiments, the yield point is improved by, for example, at least 10%, at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21%, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, or at least 50%.

[0192] In some embodiments, the articles are characterized by a structural strength where greater than 20% of the structural strength comes from incorporated fibers. In some embodiments, the compositions are characterized by a structural strength where greater than 20% of the structural strength comes from incorporated fibers. In some embodiments, the composites are characterized by a structural strength where greater than 30% of the structural strength comes from incorporated fibers.

[0193] In some embodiments, the articles are characterized by a structural strength that derives more than 1%, more than 5%, more than 10%, more than 20%, or more than 30% of its structural strength from incorporated fibers. In some embodiments, the compositions are characterized by a structural strength that derives more than 1%, more than 5%, more than 10%, more than 20%, or more than 30% of its structural strength from incorporated fibers.

[0194] In some embodiments, the phrase "structural strength" as used herein refers to mechanical properties such as, but not limited to, elastic modulus, tensile stress, elongation (strain), and toughness (e.g., a combination of tensile stress and elongation (strain)).

[0195] method According to an aspect of some embodiments of the present invention, there is provided a method for obtaining the above-mentioned fibers. In some embodiments, the method comprises: a. providing a lamin system protein at a concentration of 10 mg / mL to 400 mg / mL; and b. contacting (or injecting) the lamin system protein with a coagulation solution, thereby forming a fiber. In some embodiments, the contacting comprises an injecting step.

[0196] In some embodiments, providing the lamin system protein is at a concentration of 10 mg / mL to 400 mg / mL, 20 mg / mL to 400 mg / mL, 30 mg / mL to 400 mg / mL, 50 mg / mL to 400 mg / mL, 70 mg / mL to 400 mg / mL, 100 mg / mL to 400 mg / mL, 200 mg / mL to 400 mg / mL, 10 mg / mL to 250 mg / mL, 20 mg / mL to 250 mg / mL, 30 mg / mL to 250 mg / mL, 50 mg / mL to 250 mg / mL, 70 mg / mL to 250 mg / mL, or 100 mg / mL to 250 mg / mL (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0197] In some embodiments, the coagulation solution may be 0.45 cP to 3 cP, 0.46 cP to 3 cP, 0.47 cP to 3 cP, 0.48 cP to 3 cP, 0.5 cP to 3 cP, 0.45 cP to 2.5 cP, 0.46 cP to 2.5 cP, 0.47 cP to 2.5 cP, 0.48 cP to 2.5 cP, 0.5 cP to 2.5 cP, 0. and characterized by a viscosity of 45 cP to 2 cP, 0.46 cP to 2 cP, 0.47 cP to 2 cP, 0.48 cP to 2 cP, 0.5 cP to 2 cP, 0.45 cP to 1 cP, 0.46 cP to 1 cP, 0.47 cP to 1 cP, 0.48 cP to 1 cP, or 0.5 cP to 3 cP (including any ranges therebetween). Each possibility represents a separate embodiment of the present invention.

[0198] In some embodiments, the coagulation solution is characterized by a viscosity of greater than 0.7 cP, greater than 0.8 cP, greater than 0.9 cP, greater than 1 cP, greater than 1.5 cP, greater than 1.7 cP, or greater than 2 cP (including any value therebetween). Each possibility represents a separate embodiment of the present invention.

[0199] In some embodiments, the infusing step is performed at a flow rate of at least 0.1 mL / h, at least 0.2 mL / h, at least 0.5 mL / h, at least 0.7 mL / h, at least 1 mL / h, at least 1.7 mL / h, at least 2 mL / h, at least 5 mL / h, at least 10 mL / h, at least 15 mL / h, or at least 20 mL / h (including any value therebetween), with each possibility representing a separate embodiment of the present invention.

[0200] In some embodiments, the injecting step comprises the step of injecting a volume of 0.001 mm to 1 mm, 0.005 mm to 1 mm, 0.01 mm to 1 mm, 0.02 mm to 1 mm, 0.001 mm to 0.9 mm, 0.005 mm to 0.9 mm, 0.01 mm to 0.9 mm, 0.02 mm to 0.9 mm, 0.001 mm to 0.5 mm, 0.05 mm to 0.5 mm, 0.09 mm to 0.5 mm, 0.1 mm to 0.5 mm, 0.12 mm to 0.5 mm, 0.13 mm to 0.5 mm, 0 This is accomplished via a needle having an inner diameter of 0.15mm to 0.5mm, 0.05mm to 0.3mm, 0.09mm to 0.3mm, 0.1mm to 0.5mm, 0.12mm to 0.3mm, 0.13mm to 0.3mm, 0.15mm to 0.3mm, 0.05mm to 0.2mm, 0.09mm to 0.2mm, 0.1mm to 0.2mm, 0.12mm to 0.2mm, 0.13mm to 0.2mm, or 0.15mm to 0.2mm (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0201] In some embodiments, the coagulation solution comprises (i) an alcohol, (ii) an aqueous solution, (ii) a buffer, or any combination thereof.

[0202] In some embodiments, the alcohol is selected from methanol (MeOH), ethanol (EtOH), propanol (PrOH), isopropyl alcohol (IPA), butanol, pentanol, including any additional water-miscible alcohols (e.g., C1-C5 alcohols or C1-C3 alcohols), or any combination thereof.

[0203] In some embodiments, the coagulation solution is 50% (v / v) to 100% (v / v), 60% (v / v) to 100% (v / v), 65% (v / v) to 100% (v / v), 69% (v / v) to 100% (v / v), 70% (v / v) to 100% (v / v), 72% (v / v) to 100% (v / v), 75% (v / v )~100%(v / v), 50%(v / v)~99%(v / v), 60%(v / v)~99%(v / v), 65%(v / v)~99%(v / v), 69% (v / v)~99%(v / v), 70%(v / v)~99%(v / v), 72%(v / v)~99%(v / v), 75%(v / v)~99%(v / v), 5 0%(v / v)~98%(v / v), 60%(v / v)~98%(v / v), 65%(v / v)~98%(v / v), 69%(v / v)~98%(v / v ), 70%(v / v)~98%(v / v), 72%(v / v)~98%(v / v), 75%(v / v)~98%(v / v), 50%(v / v)~90%(v 60%(v / v)-90%(v / v), 65%(v / v)-90%(v / v), 69%(v / v)-90%(v / v), 70%(v / v)-90%(v / v), 72%(v / v)-90%(v / v), or 75%(v / v)-90%(v / v) alcohol (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0204] In some embodiments, the aqueous solution comprises a cross-linking agent. In some embodiments, the cross-linking agent is a divalent metal ion (e.g., Ca 2+ ) or its salts (such as CaCl2 and MgCl2), covalent cross-linking agents (such as glutaraldehyde, paraformaldehyde, etc.), or any combination thereof.

[0205] In some embodiments, the aqueous solution contains 1 mm to 100 mm, 2 mm to 100 mm, 3 mm to 100 mm, 4 mm to 100 mm, 5 mm to 100 mm, 10 mm to 100 mm, 30 mm to 100 mm, 50 mm to 100 mm, 1 mm to 90 mm, 2 mm to 90 mm, 3 mm to 90 mm, 4 mm to 90 mm, 5 mm to 90 mm, 10 mm to 90 mm, 30 mm to 90 mm, 50 mm to 90 mm, 1 mm to 70 mm, 2 mm to 70 mm, 3 mm to 70 mm, 4 mm to 70 mm, 5 mm to 70 mm, 10 mm to 70 mm, 30 mm to 70 mm, or 50 mm to 70 mm of crosslinker, including any range therebetween. Each possibility represents a separate embodiment of the present invention.

[0206] In some embodiments, the aqueous solution may be at 1 mM to 100 mM, 2 mM to 100 mM, 3 mM to 100 mM, 4 mM to 100 mM, 5 mM to 100 mM, 10 mM to 100 mM, 30 mM to 100 mM, 50 mM to 100 mM, 1 mM to 90 mM, 2 mM to 90 mM, 3 mM to 90 mM, 4 mM to 90 mM, 5 mM to 90 mM, 10 mM-90 mM, 30 mM-90 mM, 50 mM-90 mM, 1 mM-70 mM, 2 mM-70 mM, 3 mM-70 mM, 4 mM-70 mM, 5 mM-70 mM, 10 mM-70 mM, 30 mM-70 mM, or 50 mM-70 mM of divalent metal ion salt (including any range therebetween), with each possibility representing a separate embodiment of the present invention.

[0207] In some embodiments, the aqueous solution may be at a concentration of 1 mM to 100 mM, 2 mM to 100 mM, 3 mM to 100 mM, 4 mM to 100 mM, 5 mM to 100 mM, 10 mM to 100 mM, 30 mM to 100 mM, 50 mM to 100 mM, 1 mM to 90 mM, 2 mM to 90 mM, 3 mM to 90 mM, 4 mM to 90 mM, 5 mM -90 mM, 10 mM-90 mM, 30 mM-90 mM, 50 mM-90 mM, 1 mM-70 mM, 2 mM-70 mM, 3 mM-70 mM, 4 mM-70 mM, 5 mM-70 mM, 10 mM-70 mM, 30 mM-70 mM, or 50 mM-70 mM CaCl2 (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0208] In some embodiments, the aqueous solution is 0.1% (v / v) to 10% (v / v), 0.2% (v / v) to 10% (v / v), 0.5% (v / v) to 10% (v / v), 0.9% (v / v) to 10% (v / v), 1% (v / v) to 10% (v / v), 3% (v / v) to 10% (v / v), 5% (v / v) to 10% (v / v). , 0.1% (v / v) to 7% (v / v), 0.2% (v / v) to 7% (v / v), 0.5% (v / v) to 7% (v / v), 0.9% (v / v) to 7% (v / v), 1% (v / v) to 7% (v / v), 3% (v / v) to 7% (v / v), 5% (v / v) to 7% (v / v) of crosslinker (including any range therebetween). Each possibility represents a separate embodiment of the present invention.

[0209] In some embodiments, the method includes at least one of the steps of (i) drying the fiber and (ii) stretching the fiber. In some embodiments, (i) drying the fiber is performed after step b.

[0210] In some embodiments, the method further comprises the step (c) of contacting the fiber with a hydrophobic agent, thereby forming a coating layer on the fiber. In some embodiments, (i) the step of drying the fiber is performed after step c.

[0211] In some embodiments, the fibers obtained by the above methods do not include microfibers.

[0212] In some embodiments, the method further comprises a step (iii), preceding step a, of purifying the lamin system protein.

[0213] In some embodiments, step (iii) of purifying the lamin system protein comprises (a) solubilizing the lamin system protein in a chaotropic agent, (b) removing the chaotropic agent, and (c) contacting the lamin system protein with metal ions. In some embodiments, steps (b) and (c) comprise dialysis. In some embodiments, steps (b) and (c) comprise two independent dialysis.

[0214] general As used herein, the term "about" refers to ±10% or ±20%. Moreover, all numerical values ​​disclosed herein are approximations that encompass a variation of ±10% or ±20% from the disclosed value. It is to be understood that all numerical values ​​disclosed herein are preceded by the term "about."

[0215] The words "comprises," "comprising," "includes," "including," "having" and their variations mean "including but not limited to."

[0216] The term "consisting of" means "including and limited to."

[0217] The term "consisting essentially of" means that a composition, method, or article (e.g., the fibers of the invention, or articles processed therefrom) may include additional components, steps, and / or moieties, but only if the additional components, steps, and / or moieties do not materially change the basic and novel characteristics of the claimed composition, method, or article. Additionally, the term "consisting essentially of" is used to define an article or composition that includes the recited elements, but excludes other elements that may have essential significance to the article or composition.

[0218] The word "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any embodiment described as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments and / or to exclude the incorporation of features from other embodiments.

[0219] The term "optionally" is used herein to mean "provided in some embodiments and not provided in other embodiments." Any particular embodiment of the invention may include multiple "optional" features unless such features are inconsistent.

[0220] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0221] Throughout this application, various embodiments of the present invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as strictly limiting the scope of the present invention. Thus, the description of a range should be considered to specifically disclose all possible subranges as well as individual values ​​within that range. For example, the description of a range such as 1-6 should be considered to specifically disclose subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., as well as individual numbers within that range, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0222] Whenever a numerical range is given herein, it is meant to include any recited numbers (fractional or integer) within the given range. The phrases "range between" a first designated number and a second designated number and "range from" a first designated number to a second designated number are used interchangeably herein and are meant to include the first and second designated numbers and all fractional and integer numbers therebetween.

[0223] As used herein, the term "method" refers to manner, means, techniques and procedures for accomplishing a given task, including but not limited to, those known to those of skill in the art of chemistry, pharmacology, biology, biochemistry and medicine, or readily developed from known manners, means, techniques and procedures by those of skill in the art.

[0224] As used herein, the term "treating" includes arresting, substantially inhibiting, slowing or reversing the progression of the condition, substantially ameliorating the clinical or cosmetic symptoms of the condition, or substantially preventing the appearance of clinical or cosmetic symptoms of the condition.

[0225] It is understood that certain features of the invention that are described for clarity in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described for brevity in the context of a single embodiment may also be provided separately or in any suitable subcombination, or as appropriate in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperable without those elements.

[0226] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples. EXAMPLES

[0227] Reference is now made to the following examples, which together with the above descriptions illustrate certain embodiments of the invention in a non-limiting manner.

[0228] material and method In this study, we investigated the potential of Ce-lamin fibers by spinning Ce-lamin into fibers using a wet-spinning approach. Compared to the self-assembly approach, wet-spinning allows for the preparation of thinner fibers with better control over fiber diameter. We hypothesized that proteins with different domain architectures, when prepared under different conditions, would assemble into distinct paracrystalline networks, each with unique mechanical properties. To this end, we used three protein constructs with the following composition of domains: coiled-coil rod domain (rod-Ce-lamin, 40 kDa, SEQ ID NO: 11), rod and tail domain (rod-tail-Ce-lamin, 59 kDa, SEQ ID NO: 12), and all three domains (full-length-Ce-lamin, 64 kDa, SEQ ID NO: 10) (Figure 1A). Fibers from these constructs were synthesized using different injection flow rates and Ca in the coagulation buffer. +2We prepared nanocrystals with different concentrations and compared their mechanical properties. We also tested whether the assembly conditions influenced the structure of the quasicrystals (different organization of the protofilaments) or their organization into networks.

[0229] Expression and purification of C. elegans lamins Plasmid pET24d (Novagen) containing the lmn-1 (NC_003279.8) gene was constructed according to a previously reported method. The rod- and rod-tail-Ce-lamin genes were cloned at the N-terminal 6xHis-TEV site and C-terminal AVI tag in pET24d(+) (Novagen) to generate 6His-TEV-Ce-laminAvi. The plasmid was transformed into E. coli BL21 derivative Rosetta(DE3)plysS. Overnight bacterial cultures were diluted (1100) into fresh LB medium and grown to an OD600 of 0.5–0.9. IPTG (0.3 mM) was added and after 3 h, bacteria were harvested by centrifugation. The pellet containing the inclusion bodies was resuspended in resuspension buffer (20 mM Tris-HCl, pH 7.6, 200 mM NaCl, 1 mM EDTA) containing 1:10,000 (v / v) Calbiochem Protease Inhibitor Cocktail Set III and 1% (v / v) Tween 20. The bacterial suspension was sonicated for 10 min (3 sec on and 4 sec off) with a 65% pulse (EXLAB model BM-150) and centrifuged at 8000×g for 10 min at 4° C. The inclusion bodies were washed twice with resuspension buffer and then incubated with 20 units / mL Benzonase nuclease (Novagen, Denmark) for 30 min. The inclusion bodies were then centrifuged again at 8000×g for 10 min at 4° C. and then dissolved in urea buffer (20 mM Tris-HCl, pH 7.6, 50 mM NaCl and 6 M urea). Finally, the suspension was centrifuged at 17,000×g for 1 h at 4° C. The supernatant was then concentrated to the desired concentration by a centrifugal concentrator (30,000 kDa cutoff). A NanoDrop was used to prepare Ce-lamin solutions of the desired concentration (100 mg / mL) and ratio (260 / 280, approximately 1.1).

[0230] Assembly of C. elegans lamins into filaments Purified Ce-lamins (full-length, rod-tail, and rod) in 6 M urea-containing buffer (100 mg / mL) were dialyzed (11,000 (v / v)) against dope solution conditions (0.5 M urea, 100 mM NaCl, 25 mM Tris-HCl, pH=9, and 1 mM dithiothreitol (DTT)). After dialysis, the lamin solution was centrifuged at 17,000×g for 1 h to remove aggregates and then injected through a syringe needle (22S gauge, model 710 Hamilton® syringe) into a coagulation bath at room temperature with injection flow rates of 0.5, 1.0, and 3.5 mL / h via a syringe pump (Chemyx NanoJet syringe pump) until fibers were formed. The coagulation bath contained 25 mM Tris-HCl, pH 9.0, 20 or 50 mM CaCl2, and 1 mM DTT. Finally, the fibers were stored at room temperature in coagulation buffer containing approximately 20% 2-propanol until tensile testing. All wet fibers used in the study were stable in tubes for at least 2 years.

[0231] Hydrophobic coating for textiles The fibers from the coagulation solution were first air dried for 20 minutes. The dried fibers were immersed in the desired coating oil for times ranging from 15 minutes to 24 hours and then air dried for 20 minutes. The dried fibers were immersed in a water bath for 24 hours. The fibers were then air dried for 20 minutes before the fibers were subjected to tensile testing.

[0232] Scanning electron microscope imaging of Ce-lamin macrofibers Scanning electron microscope (SEM) imaging was performed using a Thermo Verios 460L (Thermo Fisher Scientific Inc.) instrument equipped with a field emission gun. Samples were coated with chromium prior to imaging. Secondary electron images were recorded at 3 keV and a working distance of 8.1 mm.

[0233] Preparation of Ce-lamin macrofiber sections for TEM imaging Ce-lamin-based fibers were treated sequentially with 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.35) at room temperature, postfixed with 1% OsO4 in 0.1 M cacodylate buffer, and block stained with 1% aqueous uranyl acetate for 1 h each. Samples were then dehydrated in an ethanol series and embedded in Epon / Araldite (Sigma-Aldrich, Buchs, Switzerland). Ultrathin (70 nm) sections were poststained with lead citrate and examined with a Tecnai G2 Spirit transmission electron microscope (Thermo Fisher Scientific, Eindhoven, The Netherlands) using an Orius 1000 digital camera (Gatan, Munich, Germany) at an accelerating voltage of 120 kV. Diameter ranges were measured from TEM cross-sectional images using ImageJ (Table 4). Using the same tool, the repeat lengths along individual quasicrystals (dark / black and light / white segments) were measured as the length between the two centers of the black regions, and the average repeat length was calculated.

[0234] Characterization of the mechanical properties of Ce-lamin fibers Tensile tests of individual fibers were performed at room temperature using a single-column universal testing machine (Shimadzu, AGS-x) equipped with a 100 N load cell at crosshead speeds of 0.3, 10, and 100 mm / min. Environmental conditions such as temperature (approximately 25 °C) and humidity (approximately 50%) were kept relatively constant during uniaxial tensile tests. For each combination of conditions, 5 to 10 fibers were tested. Individual fibers (0.5 cm) were attached to a cardstock paper frame and glued at both ends.

[0235] Fixing the fibers with adhesive protects them from deformation that occurs during sample mounting in the apparatus, which must be done with extreme care. Fiber length was measured using a standard ruler. Wet spun fibers were air-dried for 10 min before stretching. Full-length Ce-lamin fibers formed at an injection rate of 3.5 mL / h in a coagulation bath containing 20 mm CaCl2 were also air-dried for 6 or 24 h. Force-displacement curves were converted to stress-strain curves by dividing the force by the average cross-sectional area of ​​the fiber (assuming a circular cross-section). Importantly, after testing, fiber diameter was measured at five different locations evenly distributed along the length of each fiber using an optical microscope. The average diameter was then used to calculate the fiber cross-sectional area. Thus, the fracture stress (i.e., strength) and fracture strain were calculated as the engineering stress and strain at failure (dL / L0), respectively. Young's modulus is the slope of the stress-strain curve in the elastic linear region of the curve before the yield point. Furthermore, the strain energy at failure (i.e., toughness) was calculated by measuring the area under the stress-strain curve.

[0236] statistical analysis To analyze the effect of protein type, CaCl2 concentration in the coagulation bath, injection flow rate and crosshead speed on the mechanical properties of the fibers, multifactorial ANOVA with t-tests and F-tests (p-value < 0.05) were performed using JMP v.13. The effect of combined treatments (statistical interactions) was also analyzed.

[0237] Raman analysis Data collection The Raman system included a Horiba Lab Ram HR evolution micro-Raman system equipped with a Synapse Open Electrode CCD detector air-cooled to -60 °C. The excitation source was a 785 nm diode laser with a power of 50 mW on the sample. The laser was focused to a spot approximately 2 μm in size using a 50x objective. Measurements were performed with a 600 g mm-1 grating and a 100 μm confocal microscope hole. The exposure time was 900 s.

[0238] Data Processing In order to quantify the obtained results as accurately as possible, we have validated them by using an estimation procedure based on Monte Carlo simulations and model estimation. This method models the data as a Gaussian Mixture Model (GMM), i.e. a probability density function (pdf) that includes a fixed number of K weighted normal pdfs with different means and variances. In that framework, the pdf estimation is defined as follows:

number

[0239] Example 1 Recombinant expression of Ce-lamins in E. coli as inclusion bodies (Figure 1A) is usually accompanied by their solubilization in the presence of high concentrations of chaotropic agents such as urea at the end of the purification process (Figure 1B). Thus, the in vitro formation of paracrystals requires the removal of urea and the subsequent addition of Ca in two successive dialysis steps. +2 The addition of ions is required. To avoid the formation of aggregates in the first step, the assembly of paracrystals was performed only at low concentrations of lamins (0.1-1 mg / mL). However, to spin fibers with the wet spinning approach, a high solution viscosity is required, which can be achieved by using a high protein concentration. Therefore, the dope solution composition (0.5 M urea, 100 mM NaCl, 25 mM Tris-HCl, pH 9, and 1 mM DTT) was first optimized to ensure that Ce-lamins remained soluble at a concentration of 100 mg / mL. The dope solution was injected into a coagulation bath containing 20 mM or 50 mM CaCl2 (Tables 1, 2, and 3) at a rate of 0.5, 1, or 3.5 mL / h. [Table 1] [Table 2] [Table 3]

[0240] The wet-spinning approach rather than the dialysis-based self-assembly approach allowed us to obtain wet fibers with smaller and more uniform diameters (~170 μm). We did not detect microfibers within the fibers, as seen with the dialysis procedure. To confirm the formation of quasicrystals in the coagulation buffer, we analyzed the structure of Ce-lamin wet fibers using TEM imaging, revealing a 70 nm thick cross section. With this technique, the internal structure of wet macroscopic fibers can be visualized at the nanometer level. Despite the high Ce-lamin concentration, most Ce-lamin fibers contained a large network of quasicrystals (Figure 2A-C), with diameters ranging from 30 nm to 150 nm (Figure 2B-C and Table 4). [Table 4]

[0241] Before mounting the fibers in the tensile tester, the wet fibers were air-dried for 10 min, which caused the fiber diameter to shrink to 50-80 μm. EM images (Figure 1C-H) showed some microfiber formation after drying. Thus, drying likely caused the paracrystalline network to undergo structural reorganization due to water evaporation. However, we hypothesize that the structure of the individual paracrystalline crystals remained unchanged in the presence of moisture. Increasing the drying time beyond 10 min did not cause further shrinkage of the fibers and did not significantly affect the mechanical properties after 6 or 24 h (Table 5). [Table 5]

[0242] All dry fibers were mechanically strained at rates of 0.3, 10, and 100 mm / min using a single-column universal testing machine. The response of all Ce-Lamin fibers to tensile forces was similar to that of other IF protein-based fibers, e.g., hydrated rigid α-keratin, vimentin, and Lamin fibers assembled through a dialysis procedure. Specifically, a linear elastic region (1.4%-5%) was found up to the yield point, followed by a long plastic region until fiber failure (Figure 3). Furthermore, due to the experimental setup, Ce-Lamin fibers exhibited a wide range of mechanical properties (Tables 1, 2, and 3), which were not affected by the injection flow rate, CaCl2 concentration, or protein structure alone (p-value >0.05), but by their synergistic effect (significant statistical interaction, p-value <0.05) (Table 4). Indeed, for each protein construct, we found a combination of conditions (injection flow rate and CaCl2 concentration) that resulted in optimal mechanical performance (Figure 3). These specific conditions are hereafter referred to as optimal assembly conditions. Compared with regenerated and recombinant hagfish mucus, vimentin, and self-assembled Ce-lamin protein-based fibers, the wet-spun Ce-lamin fibers exhibited better stiffness and toughness, which was comparable to that of native silk and native hagfish mucus threads.

[0243] To study the effect of different combinations of assembly conditions on mechanical performance, we investigated how the assembly conditions affect the paracrystal structure. That is, the width of the paracrystal or the mode of protofilament association within the paracrystal may be a factor in fiber mechanics. First, we measured the paracrystal width, which ranged from 10 to 50 nm for rods, 10 to 250 nm for rod-tails, and 20 to 250 nm for full-length Ce-lamin (Table 4). For each protein construct, the width of the paracrystals in fibers formed under optimal assembly conditions (Figure 3) did not show significant differences from other assembly conditions. Thus, different paracrystal widths did not affect fiber mechanics. Second, we measured the black-and-white average repeat length along individual paracrystals. The average repeat length indicates the mode of association of dimers with protofilaments within individual paracrystals. The average repeat length of full-length and rod-tail Ce-lamin fibers was about 40 nm in all assembly conditions (Figures 2A-C and 5A-C), suggesting that protofilament association was not affected under different spinning conditions. Moreover, under most conditions, rod-Ce-lamin formed filamentous networks lacking repeat length. These networks were similar in morphology and filament diameter to quasicrystals analyzed at higher resolution using cryo-electron tomography reported in the literature. In that study, the association of protofilaments into quasicrystals was similar to that of full-length Ce-lamin, suggesting that the lack of average repeat length was probably due to the removal of the long and dense tail domain (179 amino acids), especially its Ig-fold segment. Therefore, we hypothesized that the filamentous structure of rod-Ce-lamin fibers is a thin quasicrystal. Thus, we were not able to determine whether the different assembly conditions had any effect on protofilament organization into rod-Ce-lamin fibres.

[0244] We also tested whether the diameter of the dry fibers after tensile testing could be correlated with the mechanical properties of each Ce-Lamin construct (Figure 4A-C). Continuous tension on the fibers during tensile testing at different crosshead speeds further contracted the diameter of the dry fibers (50-80 µm) to 27-61 µm (Figure 4D). The fibers with the smallest diameter could exhibit the highest stiffness and toughness properties. In the optimal assembly conditions, (Figure 3) rod-Ce-Lamin fibers showed a similar average diameter to the fibers formed in the other conditions. In contrast, the diameters of the rod-tail and full-length Ce-Lamin fiber morphologies were smaller than the fibers formed in the other conditions. However, these differences were not statistically significant (Figure 4A-C). For example, full-length Ce-Lamin fibers formed at injection rates of 1 or 3.5 mL / h in a coagulation bath containing 20 mm CaCl2 and then strained at 0.3 mm / min exhibited similar diameters on average, but completely different stresses, stiffness, and toughness. Moreover, full-length Ce-Lamin fibers assembled with the dialysis procedure were much thicker (~217 μm) compared to the wet-spun fibers (~50 μm) after tensile testing (present study). Both fibers were strained at a crosshead speed of 0.3 mm / min and exhibited similar average stresses, stiffness, and toughness. This probably means that on the one hand the fiber diameter does not affect fiber mechanics, but on the other hand it implied that if we could produce thinner fibers using the dialysis procedure, the fibers would be tougher and stiffer. Finally, a closer look at the morphology of full-length Ce-lamin fibers (Figures 1F-H) showed a more uniform structure, composed of compacted micron-sized (1-2 μm) microfibers, than rod and rod-tail fibers, which were composed of non-uniform thicker microfibers. These differences in microfiber organization and size may explain the differences in mechanical properties between the fibers at optimal conditions.

[0245] To assess whether the amount of β-sheet structure formed due to secondary structural transition upon stretching of wet-spun Ce-lamin fibers could be a factor in fiber mechanics, we applied Raman spectroscopy (Figure 5A-C). The secondary structural transition from α-helix to β-sheet, which results in fibers that are stiffer and stronger than fibers rich in elastic α-helices, is primarily responsible for the high strength and strain of spider silk and α-keratin fibers. Raman spectra (Figure 5A-C) of the toughest fibers (Figure 3) before and after mechanical testing show that the α-helix (1650 cm) is the dominant β-sheet structure. -1 ) and β-sheet and / or random coil structures (1667 cm -1 ) (Figure 5A-C). To evaluate the increase in the number of β-sheet structures as a result of the pulling process, we used an independent curve fitting method, modeling the curve as a mixture of four Gaussian distributions whose parameters were estimated using a regular EM algorithm. The results showed that the number of β-sheets increased and the number of α-helical structures decreased. The rod-based fibers showed a milder increase in β-sheet structures (4%) than that seen in rod-tail (12%) and full-length (8%) Ce-lamins. Despite these differences, we did not find any effect of the increase in β-sheet structures on the stiffness, breaking stress, and strain percentage of fibers formed in optimal conditions. However, this result may suggest that the presence of C-terminal sequences, N-terminal sequences, or both sequences promote the formation of β-sheet structures during tensile testing by forming densely packed protofilaments. We therefore speculate that the stress-strain properties of the fibers are not affected by the formation of higher β-sheet structures, fiber diameter, quasicrystal width, or protofilament association (average repeat length), but by the number of connection points between quasicrystals (which can be recognized as the number of interactions between protofilaments from two separate quasicrystals). Thus, the structural rearrangement during tensile testing may start from the unraveling of those interconnected quasicrystals, caused by the alignment and sliding of the protofilaments that connect them. This may be due to the fact that the Ca +2This can result in the destruction of intermolecular crosslinks, thereby causing irreversible deformation of the connection points between the quasicrystals, leading to a yield point. From this point, the alignment and sliding of the protofilaments and polymers of the dimers within the quasicrystals continues until the fiber breaks (Figure 6A-6B). In accordance with this, based on the Ce-lamin constructs studied here, the effect of the addition of the C-terminus or both the N- and C-terminus to the rod domain on the mechanical properties was not clear. However, at the optimal conditions, the full-length fibers showed the highest stiffness and toughness at all crosshead speeds. Furthermore, the rod and rod-tail fibers showed similar stiffness and toughness at a crosshead speed of 0.3 mm / min, but at 10 mm / min, both fibers showed better stiffness and toughness, with the rod fibers demonstrating better properties. Further increasing the speed to 100 mm / min resulted in very high stiffness and toughness of the rod-tail fibers in contrast to the rod fibers, which showed a decrease in properties. At most assembly conditions, increasing the speed resulted in higher stiffness, except for the optimal conditions for the rod and full-length fibers. In these cases, the stiffness decreased at 100 mm / min, whereas for the rod-tail fibres, the stiffness increased by 2.5-fold, likely due to the effect of the N- and C-termini in certain conditions on the binding of the coiled-coil domains in dimer formation and on the association of the two dimers, thus strengthening the dimers and protofilaments.

[0246] This study demonstrates that the recombinant IF protein, Ce-lamin, can be solubilized and wet-spun in aqueous solution to form wet macroscopic fibers (diameter ~170 μm) composed of a complex network of quasicrystals (diameter range of 10 nm to 250 nm), as revealed by TEM cross-section analysis. Our results suggested that the mechanical properties probably depend on the formation of a specific network of quasicrystals formed under specific assembly conditions. To further investigate the quasicrystal network, higher resolution structural analysis would be necessary to reveal the manner of protofilament association between two interconnected quasicrystals at different assembly conditions. Raman analysis demonstrated an increase in β-sheet structure. Potentially, Ce-lamin coiled coils can completely transform into β-sheet structure and thus withstand higher strain rates and achieve higher fracture stresses. Nevertheless, the high toughness of Ce-lamin fibers is mainly attributed to their large strain rates, which may result from the rapid reorganization and relaxation of quasicrystals, protofilament and rod domains when exposed to stress. Thus, the formation of thinner fibers, in which local defects are less likely to cause fiber fracture, could theoretically approach complete conversion.

[0247] The Ce-lamin dry fibers were characterized by a combination of high failure strain, moderate to high stiffness and strength, making them as tough as natural hagfish and spider silk fibers. Among soft biomaterial system proteins, lamin fibers exhibit unique mechanical properties. They can strain to a much higher percentage than silk, keratin, or collagen, and have similar stiffness, but they break at a lower stress. Thus, Ce-lamin can be utilized as a component of composite biomaterials in diverse applications suitable for very tough and stiff fibers. The possibility of different types of lamins from different organisms exhibiting a wide range of mechanical properties is intriguing. For example, the association mode involved in the assembly of human lamin A dimers into quasicrystals is different from that of Ce-lamin. The altered quasicrystal structure could affect the mechanics of the fibers. Another perspective would be to analyze composite fibers containing two types of lamins, such as human lamins A and B1, as found around the cell nucleus.

[0248] Example 2 Preparation of Ce-lamin fiber in alcohol solution The wet spinning method is based on an extrusion process in which a polymer solution is injected into a coagulation bath to form fibers. The properties and structure of the fibers are affected, among others, by the injection rate, the diameter of the syringe needle, the composition of the coagulation bath, and the concentration of the protein in the dope solution. Here, the effect of the coagulation bath composition on Ce-Lamin fibers was investigated, while the remaining parameters remained constant. That is, Ce-Lamin protein solutions at a concentration of 100 mg / mL were injected into different coagulation solutions at a flow rate of 1.5 mL / h through a needle with an inner diameter of 0.168 mm. We successfully utilized various alcohol-based solutions (e.g., MeOH, EtOH, PrOH, IPA) with concentrations ranging from 50% to 100% for the formation of fibers.

[0249] The strain-stress curves of all fibers obtained after tensile testing at a speed of 10 mm / min showed nonlinear mechanical behavior similar to viscoelastic materials and other intermediate filament protein-based fibers, with a short elastic region of 3-6% and long plastic deformation. Some of the tested fibers showed higher toughness and strain values ​​than Ce-lamin fibers injected in aqueous solvents. The average strain values ​​of these fibers were close to 200%.

[0250] Raman spectroscopy Raman spectroscopy was performed to investigate the secondary structure of the proteins in the fibers, for example the ratio of α-helices to β-sheets and the change in this ratio at different tensile stages. Information on the region / stage where the main transition between α-helices and β-sheets occurs could be a clue to understand which properties of the alcohol and the coagulation bath affect the mechanical properties of the fibers. In this analysis, fibers injected in 70% ethanol were tested at various stages shown in Figure 7A. In each test, a curve of intensity versus wavenumber was obtained (Figure 8). The area under each curve represents the relative amount of secondary structure and, by statistical calculation, new curves can be constructed, one of which represents the total amount of α-helices and the other the total amount of β-sheets. As shown in Figure 7A, the transition from α-helices to β-sheets mostly occurs in the elastic phase, since up to a strain of 6% (just after the yield point (about 5%)), a decrease in the amount of α-helices and an increase in the amount of β-sheets can be seen. After a strain of 6%, there is no significant change in the percentage of the two different structures.

[0251] Raman analysis was performed on different fibers for each strain percentage, demonstrating that the transition propagation was different in fibers assembled in 70% ethanol (7A) and aqueous buffer containing 20 mM CaCl2 (7B). The relative amount of α-helix to β-sheet structure was similar at 0% strain and breakage in both conditions. However, a significant portion of the α-β transition for fibers assembled in 70% ethanol occurred up to 6% strain and remained almost unchanged until breakage. In contrast, for fibers fabricated in aqueous buffer, the relative amount of α-helix to β-sheet remained relatively constant up to 20% strain (see Figure 7B).

[0252] Furthermore, we subjected the fibers obtained from the alcohol and calcium chloride solutions to cyclic strain tests. Surprisingly, two significant differences between the assembly conditions were observed: Fibers assembled in 1.70% ethanol exhibited significant hysteresis in the five-cycle test (Figure 12B), whereas CaCl2-coagulated fibers exhibited nearly linear elastic behavior, as reflected by nearly identical strain / stress curves (Figure 12A). 2. Raman spectroscopy (Figure 12C-D) shows that when the five-cycle test was terminated in the extended position (Figure 12C), more α-helices were present. That is, there was no transition in the secondary structure during the cycle test. In contrast, for fibers assembled in 70% ethanol, the cycle test was terminated in the relaxed position (Figure 12D), showing more β-sheet / random coil structures. Thus, during the five-cycle test, an irreversible transition from α-helices to β-sheet / random coil occurred in the fibers obtained via alcoholic coagulation.

[0253] Scanning electron microscope (SEM) analysis Using SEM analysis, we investigated the surface morphology of dried Ce-Lamin fibers assembled in 50% and 70% ethanol and IPA. Figures 9A-9D and Figures 10A-10C show the surfaces of fibers that were not or were subjected to tensile load, respectively. Before stretching, all fibers were composed of large clusters of nanometric fibers that constitute the structural units of macroscopic fibers. Only in IPA do the nanofibers associate into microfibers that together constitute the macroscopic fibers. After stretching, the fine fiber organization in the IPA fibers disappears. Measurement of the nanofiber diameter showed that in 50% alcohol, the fiber diameter was smaller than in both ethanol and IPA at 70% (Table 6). The difference in diameter was an average of 16 nm in ethanol and 13 nm in IPA. Before stretching, the surface morphology of dried Ce-Lamin fibers contained large clusters of nanofilaments that constitute the structural units of macroscopic fibers. The diameter of nanofilaments assembled in 50% alcohol was approximately 15 nm lower than either 70% ethanol or IPA. These differences may explain the lower mechanical strength of fibers formed in 50% alcohol. [Table 6] [Table 7]

[0254] film The inventors further succeeded in forming transparent films (e.g., hydrogel films) composed of full-length lamin-based protein fibers of the present invention (data not shown). Both fibers obtained from aqueous coagulation and alcoholic coagulation were successfully carried out for film formation using a dialysis procedure in 70% ethanol or in an aqueous buffer containing 20 mm CaCl2, respectively. The resulting films exhibited the desired mechanical strength (e.g., about 300% breaking strain). Furthermore, Q159K mutated into Ce-lamin fibers has been carried out for the formation of transparent films. The mechanical properties of the films are summarized in Tables 7A-B below.

[0255] [Table 8] [Table 9]

[0256] Furthermore, the inventors are currently conducting various experiments to test the compatibility of the fibers of the present invention with additional polymer matrices to obtain reinforced composite materials. The inventors are testing various additional synthetic and / or natural and / or biodegradable polymers (such as those disclosed above) to produce composites with different constants (such as those disclosed herein) of the fibers of the present invention. The composites can be prepared by casting, melting, extrusion, dipping, coating, or any other method known in the art of polymer processing.

[0257] To this end, the inventors have successfully formed epoxy resin-based composites and have demonstrated excellent compatibility of the fibers of the present invention with at least synthetic resins (e.g., thermosetting resins). It is therefore hypothesized that additional polymers are compatible with the fibers of the present invention, allowing the formation of composites with various contents of the fibers of the present invention.

[0258] Hydrophobic coating on Ce-lamin fibers In nature, the protein core of silk and hair fibers is coated with a hydrophobic layer. This layer does not directly contribute to the mechanical properties of the fiber, but it protects the protein fiber from hydration and external damage. The fiber can be coated with lubricants or finishes such as polymer or wax finishes including, but not limited to, mineral oil, fatty acids, isobutyl stearate, tallow fatty acid 2-ethylhexyl ester, polyol carboxylate esters, coconut oil fatty acid esters of glycerol, alkoxylated glycerol, silicone, dimethylpolysiloxane, polyalkylene glycol, polyethylene oxide, and propylene oxide copolymer. We coated fibers assembled in 70% ethanol with paraffin oil for different immersion times to check their effect on fiber mechanics. At overnight (ON) immersion time, the coated Ce-lamin fiber was much stronger and strained to a higher percentage (Table 8), and therefore exhibited very high toughness (approximately 600 MJ / m 3 ), and the strain at break was significantly increased by about 400% and even more compared to the uncoated fiber. [Table 10]

[0259] Additionally, we tested the swelling behavior of coated versus uncoated fibers. Coated with paraffin oil (A) or uncoated (B), Ce-Lamin fibers were immersed in water for 4 days. Fiber swelling was observed only for the uncoated fibers, demonstrating the water-repellent (or superhydrophobic) effect of the hydrophobic coating.

[0260] We subjected the coated fibers to cyclic strain tests. The five-cycle experiment demonstrated the linear elastic behavior of the coated fibers. Raman spectroscopy analysis shows that when the five-cycle test ends in the stretched position, the fibers have more beta sheets / random coils and less alpha helices compared to the relaxed position, which has more alpha helices. Thus, a reversible transition from alpha helices to beta sheets / random coils occurred during the five-cycle test.

[0261] Example 3 Human lamin A fiber The unique environment of lamins within the cell nucleus allows A-type and B-type lamin filaments, each forming a distinct fiber meshwork, to form biocomposite materials with the nuclear membrane, lamin-associated proteins, and chromatin, which apparently affect their structure and mechanics. Here, we expressed A-type human lamin (corresponding to the amino acid sequence of SEQ ID NO: 9) and wet-spun them into three different coagulation baths: 1. 70% ethanol in water, 2. Tris buffer (20 mM Tris-pH-9, 20 mM CaCl2), 3. MES buffer (50 mM MES-pH-6, 10 mm CaCl2). Bacterial expression, protein purification, and dope solution preparation were performed as described in the Ce-Lamin procedure.

[0262] Overall, human lamin A-based fibers had the same stress-strain behavior as Ce-lamin fibers, but lower toughness and strength (Table 8A). [Table 11]

[0263] Example 4 Mutant Ce-lamin fibers The Q159K mutation in Ce-lamin is similar to the E145K mutation in human lamin A, one of the causes of progeria. The amino acid glutamine is substituted with lysine at position 159. The mutant protein solution was injected into a coagulation bath containing 70% ethanol and 70% IPA. The mutation significantly affected the yield and break stresses, break strain, and toughness, while Young's modulus and fiber diameter were not significantly changed (Figure 11 and Table 9). The average toughness of the native fibers was three times that of the mutant fibers in ethanol and more than two times that in IPA. Although the mutant lamin protein-based fibers are characterized by a reduced toughness, the fibers are suitable for industrial applications, especially those that do not require excessive mechanical strength.

[0264] Furthermore, it is hypothesized that the physical properties (e.g., mechanical strength and / or Young's modulus) of the various mutants can be adjusted to obtain fibers with tailored physical properties. Such fibers may be useful in a variety of composite materials where similar physical properties (e.g., Young's modulus) of the components are required. [Table 12]

[0265] While the present invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications, and variations that fall within the spirit and broad scope of the appended claims.

[0266] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated herein by reference.Furthermore, citation or identification of any reference in this application should not be construed as an admission that such reference is available as prior art to the present invention.To the extent section headings are used, they should not be construed as necessarily limiting.

Claims

1. 1. A fiber comprising a lamin family protein, said fiber being characterized by an average diameter of 1 μm to 1000 μm and an average length of at least 1 mm, said fiber being covered by a coating.

2. The fiber of claim 1 , wherein the hydrophobic coating is in the form of a layer and comprises a water-immiscible compound.

3. the water-immiscible compound is selected from vegetable oils, mineral oils, fatty acids, fatty acid esters, lipids, isobutyl stearate, tallow fatty acid 2-ethylhexyl esters, polyol carboxylic acid esters, glycerides, coconut oil fatty acid esters of glycerol, alkoxylated glycerol, silicone oils, dimethylpolysiloxanes, and waxes, including any copolymers, any salts, or any combination thereof; The layer is characterized by an average thickness of 1 nm to 20 μm. The fiber of claim 2.

4. 2. The fiber of claim 1, wherein the lamin family protein undergoes an alpha helix to beta sheet transition upon stretching the fiber to about 6% strain.

5. A fiber comprising a lamin system protein, said fiber characterized by an average diameter of about 10 μm to about 1000 μm, said lamin system protein undergoing an alpha helix to beta sheet transition upon stretching said fiber to a strain of about 6%.

6. the fibers have an average diameter of about 10 μm to 180 μm, and the α-helix to β-sheet transition comprises at least a 20% α-helix to β-sheet transition as determined by Raman spectroscopy; the fibers are characterized by an average length of at least 1 nm; When the fibers are stretched to strains of about 6% to about 50%, the lamin family proteins are characterized by a β-sheet content of about 30% to about 50%. The fiber of claim 5.

7. the lamin system protein comprises an A-type lamin, a B-type lamin, or both; the B-type lamin comprises the amino acid sequence set forth in SEQ ID NO: 1, wherein X 1 comprises Gln or Lys, and this amino acid sequence includes any functional analogues having at least 70% sequence identity thereto; the A-type lamin comprises the amino acid sequence set forth in SEQ ID NO: 4 or SEQ ID NO: 7, wherein X 2 comprises Glu or Lys, and this amino acid sequence includes any functional analogues having at least 70% sequence identity thereto; The lamin system protein is arranged within the fiber in the form of paracrystals, each of the paracrystals being characterized by a dimension selected from: (i) a width of 1 nm to 500 nm; (ii) a length of 0.5 mm to 1 cm, or both of (i) and (ii). The fiber of claim 5.

8. 8. The fiber of claim 7, further comprising either: (i) an N-terminal region comprising an amino acid sequence set forth in SEQ ID NO: 2, including any functional analogues thereof having at least 70% sequence identity thereto; and (ii) a C-terminal region comprising an amino acid sequence set forth in SEQ ID NO: 3, including any functional analogues thereof having at least 70% sequence identity thereto.

9. The fiber described in claim 7, wherein the amino acid sequence of the A-type lamin further comprises either (i) an N-terminal region comprising the amino acid sequence shown in SEQ ID NO: 5; and (ii) a C-terminal region comprising the amino acid sequence shown in SEQ ID NO: 6 or the amino acid sequence shown in SEQ ID NO:

8.

10. The fibers are At least one mechanical property selected from the following: a yield strength of about 1 MPa to about 1000 MPa; a tensile strength of about 1 MPa to about 1000 MPa; - strain at break of about 80% to about 1000%; -About 30MJ / m ３ ~Approx. 1000MJ / m ３ Toughness and Young's modulus of about 0.001 GPa to about 30 GPa The fiber of claim 5 characterized by:

11. A fiber comprising a B-type lamin system protein, said fiber (i) characterized by a diameter of 1 μm to 1000 μm; and (ii) said B-type lamin system protein comprises amino acids set forth in SEQ ID NO: 1, including any functional analogues having at least 70% sequence homology thereto; X １ is Gln, the fiber has (i) a residual amount of alcohol; (ii) about 190 MJ / m ３ and (ii) the protein is characterized by an α-helix to β-sheet transition upon stretching it to a strain of about 6%.

12. characterized by an average diameter of about 30 to about 60 μm; (i) an N-terminal region comprising the amino acid sequence SEQ ID NO:2 or any functional analog having at least 70% sequence identity thereto; (ii) a C-terminal region comprising the amino acid sequence of SEQ ID NO: 3 or any functional analog having at least 70% sequence identity thereto; Further includes any of The fiber of claim 11.

13. The B-type lamin system protein is arranged within the fiber in the form of quasicrystals, each of the quasicrystals being characterized by dimensions selected from: (i) a width of 1 nm to 500 nm; (ii) a length of 0.5 mm to 1 cm, or both of (i) and (ii); the fiber is an elongated fiber characterized by a secondary structure comprising an alpha helix:beta sheet ratio of 5:1 to 1:1, At least one mechanical property selected from the following: a yield strength of about 1 MPa to about 1000 MPa; a tensile strength of about 1 MPa to about 1000 MPa; - strain at break of about 100% to about 1000%; -About 30MJ / m ３ ~Approx. 1000MJ / m ３ Toughness and Young's modulus of about 0.001 GPa to about 30 GPa characterized by, The fiber of claim 11.

14. An article comprising a plurality of the fibers of claim 1.

15. 15. The article of claim 14, in the form of a yarn, a mesh, a woven substrate, a nonwoven substrate, a composite material, or any combination thereof.

16. A method for obtaining the fibers according to claim 5, comprising the steps of: a. providing a lamin system protein at a concentration of 10 mg / mL to 400 mg / mL; b. injecting the lamin-based protein into a coagulation solution comprising alcohol, thereby forming the fiber; A method comprising:

17. the coagulation solution is characterized by a viscosity of 0.45 cP to 3 cP, or greater than 0.7 cP; the injecting step is performed at a flow rate of at least 0.1 ml / h; The alcohol is selected from methanol (MeOH), ethanol (EtOH), propanol (PrOH), isopropyl alcohol (IPA), butanol, pentanol, or any combination thereof.

17. The method of claim 16.

18. 17. The method of claim 16, wherein the coagulation solution comprises from 50% (v / v) to 100% (v / v) of the alcohol.

19. 17. The method of claim 16, further comprising at least one of the steps of: (i) drying the fibers; and (ii) stretching the fibers.

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