De novo engineered polypeptides and methods of use thereof

Abstract

The disclosure provides de novo designed and synthesized non-naturally occurring, beta strand reservoir polypeptides that can be processed into a beta sheet crystalline phase. The disclosure further provides compositions and beta sheet crystalline phases comprising the disclosed proteins, and methods of preparation and use thereof.

Description

[0001] Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0002] DE NOVO ENGINEERED POLYPEPTIDES AND METHODS OF USE THEREOF

[0003] CROSS -REFERENCE TO RELATED APPLICATION

[0004] [1] The present Application claims the benefit of priority to U. S. Provisional Application No.

[0005] 63 / 760,072, filed on February 18, 2025, the contents of which is hereby incorporated by reference in its entirety for all purposes.

[0006] REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0007] [2] The contents of the electronic sequence listing (ARZE_046_01WO_SeqList_ST26.xml; Size: 2,803,167 bytes; and Date of Creation: February 17, 2026) are herein incorporated by reference in its entirety.

[0008] BACKGROUND

[0009] [3] Renewable and biodegradable polymers are of increasing interest as an alternative to petroleum-based products. To this end, considerable effort has been made to develop methods of making polymers, such as fibers, from molecules derived from plants and animals. Naturally occurring fiber-forming proteins, such as silk fibroin, have unique mechanical properties, such as tensile strength, elasticity, and biocompatibility, making them valuable for a wide range of applications, including textiles, biomedical devices, and regenerative medicine. Natural sources of fiber-forming proteins are limited by availability, ethical concerns, and environmental impact. Importantly, the extraction and purification of these naturally occurring proteins is labor-intensive and costly, hindering large-scale production and commercialization.

[0010] [4] There is a need for non-naturally occurring polypeptides that have the capability to form biodegradable fibers. There are several challenges in generating such polypeptides. First, de novo protein engineering is an uphill task, involving protein structure prediction and optimization of protein folding and expression. Second, there are challenges related to achieving the desired protein yield, functionality and post-translational modifications. Finally, the proteins must be amenable to forming fibers having the desired mechanical characteristics.

[0011] SUMMARY

[0012] 1

[0013] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0014] [5] The disclosure provides, for instance, a beta strand reservoir polypeptide, comprising multiple beta domains and a linker between each of the beta domains, wherein each of the beta domains comprises an amino acid sequence having at least about 75% identity to any one of SEQ IDNOs: 91-96.

[0015] [6] The disclosure provides, for instance, a beta strand reservoir polypeptide comprising multiple beta domains and a linker between each of the beta domains, wherein the beta domains associate with each other in silico to form an octameric structure having D4 dihedral symmetry or a dodecameric structure having D6 dihedral symmetry, and each of the beta domains comprises about 40 amino acid residues to about 100 amino acid residues and exhibits an average pLDDT of over 50 (e.g. over 70).

[0016] [7] The disclosure provides, for instance, a beta strand reservoir polypeptide comprising at least four beta strands and a linker between each of the beta strands, wherein the beta strand reservoir polypeptide comprises a beta solenoid motif. In embodiments, the beta solenoid motif comprises a consensus amino acid sequence of any one of SEQ ID NOs: 1590-1600, and 1602-1655. Optionally, the at least four beta strands associate to form two parallel beta sheets (Bl and B2) in silico. In embodiments, the beta strand reservoir polypeptide comprises at least ten beta strands, optionally, wherein the at least ten beta strands associate to form two parallel beta sheets (Bl and B2) in silico.

[0017] [8] The disclosure provides, for instance, a beta strand reservoir polypeptide comprising at least four beta strands and a linker between each of the beta strands. In embodiments, the at least four beta strands associate to form two parallel beta sheets (Bl and B2), and the polypeptide exhibits a beta solenoid structure in silico.

[0018] [9] The disclosure provides, for instance, a beta strand reservoir polypeptide comprising at least ten beta strands and a linker between each of the beta strands. In embodiments, the at least ten beta strands associate to form two anti-parallel beta sheets (Bl and B2) in silico.

[0019]

[0010] The disclosure provides, for instance, a beta strand reservoir polypeptide comprising an amino acid sequence having at least about 75% identity to an amino acid sequence of any one of SEQ ID NOs: 1-84.

[0020] 2

[0021] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0022]

[0011] The disclosure provides, for instance, a beta sheet crystalline phase comprising a plurality of beta strand reservoir polypeptides, wherein the plurality of beta strand reservoir polypeptides comprises at least one beta strand reservoir polypeptide with an amino acid sequence having at least about 75% identity to any one of amino acid sequences of SEQ ID NOs: 1-84.

[0023]

[0012] The disclosure provides, for instance, a fiber comprising a beta sheet crystalline phase. In embodiments, the polymers (e.g. polypeptides) comprised by the beta sheet crystalline phase comprise less than 60% alanine and glycine and the average molecular weight of the polymers (e.g. polypeptides) is greater than 20 kDa.

[0024]

[0013] The disclosure provides, for instance, a fiber, fibrous article, or non-fibrous article comprising polymers. In embodiments, the polymers comprise a polypeptide block copolymer of the formula (B-O)n, where n is 2 or greater, e.g, between 2 and 40, B is each independently a block, greater than 16 amino acids in length, predominantly comprising beta strands, and O is each independently a block predominantly comprising non-beta strand structure (e.g., amorphous or helical regions). In embodiments, the polymers include less than 60% or less than 50% alanine and glycine. In embodiments, the polymers are characterized by having a first peak corresponding to molecular vibrations in its infrared frequency spectra at approximately 1624 cm-1 (e.g., 1624 cm-1 or 1628 cm-1) and a second peak corresponding to molecular vibrations in its infrared frequency spectra at approximately 1640cm- 1 (e.g., 1640cm- 1 or 1641 cm-1), wherein the ratio of the first peak area divided by the second peak area is greater than 1. In embodiments, the polymers have an average molecular weight of greater than 20 kDa. As used herein, “predominantly comprising” refers to “comprising more than 50%. ” For instance, “predominantly comprising beta strands refers to “comprising more than 50% beta strands.”

[0025]

[0014] The disclosure provides, for instance, a fiber, fibrous article, or non-fibrous article comprising polymers wherein the mean initial modulus of the fiber is greater than 5 GPa (e.g., greater than 10 GPa or greater than 15 GPa). In embodiments, the polymers have an average molecular weight of greater than 20 kDa, and the polymers comprise a block copolymer comprising B and O blocks. In embodiments, the B blocks predominantly comprise beta strands; the O blocks predominantly comprise non-beta strand structure (e.g., amorphous or helical regions), and the blocks B and O alternate. In embodiments, the copolymer comprises at least 2

[0026] 3

[0027] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0028] occurrences of the B block, the O blocks each independently comprises at least 15 amino acid residues, the B blocks each independently comprise at least 16 consecutive amino acids with no more than 3 consecutive amino acids comprising alanine, and the B blocks comprise no more than 34% alanine and glycine, no more than 15% glycine, and no more than 19% alanine.

[0029]

[0015] The disclosure provides, for instance, a fiber comprising a plurality of non-naturally occurring beta sheet crystalline phases, wherein each of the non-naturally occurring beta sheet crystalline phases comprises two or more beta sheets and a non-beta strand structure (e.g., amorphous or helical regions) between each of the beta sheets. In embodiments, the fiber has a tensile strength greater than or equal to 120, 130, or 140 MPa and was produced from beta strand reservoir polypeptides with a molecular weight of greater than 20 kDa and whose in silico structures comprise a plurality of beta sheets that each have an average pLDDT of over 50 (e.g. over 70).

[0030]

[0016] The disclosure provides, for instance, a nanofiber web comprising a beta sheet crystalline phase, wherein the beta sheet crystalline phase comprises a plurality of beta strands, the polymers (e.g. polypeptides) comprised by the beta sheet crystalline phase comprise less than 60% alanine and glycine, and the average molecular weight of the polymers (e.g. polypeptides) is greater than 20 kDa.

[0031]

[0017] The disclosure provides, for instance, a film comprising a beta sheet crystalline phase, wherein the beta sheet crystalline phase comprises a plurality of beta strands, the polymers (e.g. polypeptides) comprised by the beta sheet crystalline phase comprise less than 60% alanine and glycine, and the average molecular weight of the polymers (e.g. polypeptides) is greater than 20 kDa.

[0032]

[0018] The disclosure provides, for instance, a coating comprising a beta sheet crystalline phase, wherein the beta sheet crystalline phase comprises a plurality of beta strands, the polymers (e.g. polypeptides) comprised by the beta sheet crystalline phase comprise less than 60% alanine and glycine, and the average molecular weight of the polymers (e.g. polypeptides) is greater than 20 kDa.

[0033]

[0019] The disclosure provides, for instance, a beta sheet crystalline phase, wherein the beta sheet crystalline phase comprises a plurality of beta strands, the polymers (e.g. polypeptides) comprised 4

[0034] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0035] by the beta sheet crystalline phase comprise less than 60% alanine and glycine, and the average molecular weight of the polymers (e.g. polypeptides) is greater than 20 kDa.

[0036]

[0020] The disclosure provides, for instance, a textile comprising a beta sheet crystalline phase, wherein the beta sheet crystalline phase comprises a plurality of beta strands, the polymers (e.g. polypeptides) comprised by the beta sheet crystalline phase comprise less than 60% alanine and glycine, and the average molecular weight of the polymers (e.g. polypeptides) is greater than 20 kDa.

[0037]

[0021] The disclosure provides, for instance, a textile comprising a plurality of fibers, wherein each fiber comprises a plurality of non-naturally occurring beta sheet crystalline phases, wherein each of the non-naturally occurring beta sheet crystalline phases comprises two or more beta sheets and a non-beta strand structure (e.g., amorphous or helical regions) between each of the beta sheets. In embodiments, the fibers have a tensile strength greater than or equal to 120, 130, or 140 MPa and were produced from beta strand reservoir polypeptides with a molecular weight of greater than 20 kDa and whose in silico structures comprise a plurality of beta sheets that each have an average pLDDT of over 50 (e.g. over 70).

[0038]

[0022] The disclosure provides, for instance, a plurality of protein fibers twisted around a common axis, wherein the mean initial modulus of the fibers of the plurality is greater than 5 GPa (e.g., greater than 10 GPa or greater than 15 GPa), and the fibers of the plurality comprise polymers that have an average molecular weight of greater than 20 kDa. In embodiments, the polymers comprise a block copolymer comprising B and O blocks, the B blocks predominantly comprise beta strands, the O blocks predominantly comprise non-beta strand structure (e.g., amorphous or helical regions), and the blocks B and O alternate. In embodiments, the copolymer comprises at least 2 occurrences of the B block, the O blocks each independently comprises at least 15 amino acid residues, the B blocks each independently comprise at least 16 consecutive amino acids with no more than 3 consecutive amino acids comprising alanine, and the B blocks comprise no more than 34% alanine and glycine, no more than 15% glycine, and no more than 19% alanine.

[0039]

[0023] The disclosure provides, for instance, a fiber comprising a non-naturally occurring polypeptide, wherein the polypeptide comprises a plurality of beta strands and has a molecular

[0040] 5

[0041] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0042] weight of greater than 20 kDa, and wherein the fiber is substantially free of e.g., free of) formic acid.

[0043]

[0024] The disclosure provides, for instance, a process for evaluating the fiber forming or spinnability of a protein solution that involves a) dissolving the protein in a volatile organic solvent, b) placing a droplet of no more than 5ul of said solution on a flat fluorinated film surface in a convective air stream such that the solution concentration increases over time, c) bringing a rod into contact with the surface and withdrawing the rod while observing fiber formation optically or via a force transducer, d) repeating the above process for 15 minutes or until the liquid has evaporated or an observed fiber is formed, e) noting the presence or absence of the formed fiber.

[0044]

[0025] The disclosure provides, for instance, a cosmetic composition in a cosmetically acceptable medium for the treatment or enhancement of at least one of keratin-containing materials, mucosa, or teeth, the cosmetic composition comprising: (a) an effective amount of a non-silk beta strand reservoir polypeptide or beta strand reservoir polymer, wherein said polypeptide or polymer: comprises at least 2 occurrences of a beta strand B block that predominantly comprises beta-strand structure and no more than 34% combined alanine and glycine, comprises O blocks that predominantly comprise non-beta-strand structure, and (b) at least one cosmetically acceptable auxiliary or excipient.

[0045]

[0026] The disclosure provides, for instance, A cosmetic composition in a cosmetically acceptable medium for the treatment or enhancement of at least one of keratin-containing materials, mucosa, or teeth, the cosmetic composition comprising polymers wherein the polymers comprise a block copolymer comprising B and O blocks; the B blocks predominantly comprise beta strands; the O blocks predominantly comprise non-beta strand structure (e.g., amorphous or helical regions); the blocks B and O alternate; the copolymer comprises at least 2 occurrences of the B block; and the B blocks comprise no more than 34% alanine and glycine, no more than 15% glycine, and no more than 19% alanine, and (b) at least one cosmetically acceptable auxiliary or excipient.

[0046]

[0027] The disclosure provides, for instance, a method of producing a cosmetic composition comprising the steps of:(a) providing an aqueous solution comprising a non-silk beta strand reservoir polypeptide; (b) mixing said beta strand reservoir polypeptide with one or more cosmetically active compounds or effect substances; (c) optionally inducing self-assembly or 6

[0047] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0048] aggregation of said beta strand reservoir polypeptide to form stabilized structures or particles containing the one or more cosmetically active compounds; and (d) combining the resulting mixture or particles with a cosmetically acceptable carrier or auxiliary to form a cosmetic composition.

[0049] BRIEF DESCRIPTION OF THE DRAWINGS

[0050]

[0028] The accompanying figures, which are incorporated herein and form a part of the specification, illustrate some, but not the only or exclusive, example embodiments and / or features. It is intended that the embodiments and figures disclosed herein are to be considered illustrative rather than limiting.

[0051]

[0029] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0052]

[0030] FIG. 1A depicts a schematic representation of the design and construction of certain beta strand reservoir polypeptides (BSRPs) disclosed herein and their processing to form a beta sheet crystalline phase. The blue arrows depict beta strands. The left panel shows that alternating beta strands and linkers form beta domains, which are linked to each other using polypeptide linkers and are determined to fold into a tertiary structure with internal D4 dihedral pseudo-symmetry based on in silico prediction and analysis. The right panel shows that at least 4 beta strands alternating with linkers are determined to fold into a beta solenoid structure based on in silico analysis. The BSRPs of either structure can denature and refold under suitable processing conditions to form a beta sheet crystalline phase. The beta sheet crystalline phases disclosed herein may be determined to have a structure that is crystalline as illustrated further below. FIG. 1B shows a zoomed-in schematic representation of a tertiary structure with internal D4 dihedral pseudo-symmetry based on in silico analysis of an exemplary beta strand reservoir polypeptide disclosed herein. FIG. 1C shows a zoomed-in schematic representation of a tertiary beta solenoid structure based on in silico analysis of an exemplary beta strand reservoir polypeptide disclosed herein. FIG. 1D shows a schematic representation of an exemplary embodiment of the disclosure. BSRPs are determined to exhibit a tertiary structure having more intramolecular interactions (as compared to intermolecular interactions) based on in silico analysis, such as, polypeptides with a 7

[0053] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0054] structure having internal dihedral pseudo-symmetry or a beta solenoid structure and can be processed using the methods disclosed herein to form a beta sheet crystalline phase with a quaternary structure having more intermolecular interactions (as compared to intramolecular interactions). In the panel on the right, for clarity, a single polypeptide chain is traced with a red outline, highlighting how individual polypeptide chains span multiple parts of the beta sheet crystalline phase and participate predominantly in intermolecular beta strand interactions as opposed to intramolecular beta strand interactions.

[0055]

[0031] FIG. 2A shows a schematic representation of the predicted quaternary structure exhibited by a beta sheet crystalline phase formed by an exemplary beta strand reservoir polypeptide disclosed herein. A single polypeptide is highlighted in red. FIG. 2A shows that the polypeptides are stretched linearly along the axis of the quaternary structure leading to the formation of more intermolecular interactions. A plurality of beta strands, comprising one or more distinct amino acid sequences, form beta sheets exhibiting parallel or antiparallel orientation. FIG. 2B shows a schematic representation of a crystallite structure formed by a stack of beta sheets (indicated by progressively lighter shading of the beta sheets beneath the top (dark shaded) beta sheet).

[0056]

[0032] FIGs. 3A-3D shows a schematic representation, based on in silico analysis, of an octameric structure with D4 dihedral symmetry formed by eight beta domains from an exemplary non-naturally occurring BSRP disclosed herein, pA55743. In FIGs.3A and 3B, the polypeptide regions are colored by their pLDDT values, which indicates a high local confidence of the polypeptide structure determined by ColabFold. In FIGs. 3B and 3D, the main (4-fold) symmetry axis is indicated by a green arrow. In FIGs. 3C and 3D, the schematic structures are colored to indicate loop, beta strand and helix, and the Ca carbon of the first and last residue of each domain are shown as spheres.

[0057]

[0033] FIGs. 4A-4B show a schematic representation, based on in silico analysis, of an octameric structure with D4 dihedral symmetry formed by eight beta domains from an exemplary non-naturally occurring BSRP disclosed herein, pA55743, with each of the eight beta domains in a different color. The Ca carbon of the first and last residue of each domain are shown as spheres.

[0058]

[0034] FIGs. 5A-5B show a schematic representation of an exemplary non-naturally occurring BSRP disclosed herein, pA57501, which is determined to fold into a beta solenoid structure. In 8

[0059] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0060] FIG. 5A, the polypeptide regions are colored by their pLDDT values, which indicates the local confidence of the polypeptide structure determined by ColabFold. In FIGs. 5B the schematic structure is colored to indicate linkers, beta solenoid motifs and capping domains.

[0061]

[0035] FIGs. 6A-6B show a schematic representation of an exemplary non-naturally occurring BSRP disclosed herein, pA56621 which is determined to fold into a beta solenoid structure. In FIG. 6A, the polypeptide regions are colored by their pLDDT values, which indicates the local confidence of the polypeptide structure determined by ColabFold. In FIG. 6B, the schematic structures are colored to indicate linkers, beta solenoid motifs and capping domains.

[0062]

[0036] FIGs. 7A-7B show a schematic representation of an exemplary non-naturally occurring BSRP disclosed herein, pA57251, which is determined to fold into a beta solenoid structure. In FIG. 7A, the polypeptide regions are colored by their pLDDT values, which indicates the local confidence of the polypeptide structure determined by ColabFold. In FIG. 7B, the schematic structures are colored to indicate linkers, beta solenoid motifs and capping domains.

[0063]

[0037] FIGs. 8A-8B show schematic representation of the BSRPs disclosed herein in with a beta solenoid structure (type #1, as described below). FIG. 8A shows exemplary domains of an exemplary beta solenoid BSRP. The lighter blue arrows represent beta strands that form the first beta sheet (Bl) while the darker blue arrows represent beta strands that form the second beta sheet (B2). The C terminus of the beta strand of Bl is covalently linked to the N terminus of the beta strand of B2 via a loop (blue). The C terminus of the beta strand of B2 is covalently linked to the N terminus of the beta strand of Bl via a linker. The dotted rectangle in FIG. 8B shows the beta solenoid motif of an exemplary beta solenoid BSRP disclosed herein. FIGs. 8C-8L depict other types of beta solenoid structures that may be exhibited by BSRPs. FIGs. 8C and 8D depict a type #2 beta solenoid comprising two beta sheets connected via loops. FIGs. 8E and 8F depict a type #3 beta solenoid comprising two type #2 solenoids connected via a linker. FIGs.8G and 8H depict a type #4 beta solenoid comprising two beta sheets connected via loops except when they are connected via a linker after odd number revolutions (e.g. 3 or 5 revolutions) of the beta solenoid.

[0064] FIGs. 8I and 8J depict a type #5 beta solenoid in which the two beta sheets are connected via a linker comprising or consisting of an amino acid sequence of 8 amino acids or less, that is at least 60% glycine (such as SEQ ID NO: 147 or 148). FIGs. 8K and 8L depict a type #6 beta solenoid

[0065] 9

[0066] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0067] comprising two beta sheets connected, in an alternating manner, via (i) a linker comprising or consisting of an amino acid of 8 amino acids or less, that is at least 60% glycine (such as SEQ ID NO: 147 or 148), and (ii) a linker not comprising or consisting of the amino acid sequence of SEQ ID NO: 147 or SEQ ID NO: 148. Capping domains are present in all BSRPs disclosed herein and the structures shown in FIGs. 8D, 8F, 8H, 8J, and 8L, but are omitted from the schematics in FIGs. 8A, 8B, 8C, 8E, 8G, 8I, and 8K due to the large variety of secondary structure compositions they may comprise.

[0068]

[0038] FIG. 9A shows the tertiary structure based on in silico analysis of the BSRP disclosed herein that comprises the amino acid sequence of SEQ ID NO: 197 (pA55743_2). FIG. 9B shows the tertiary structure based on in-silico analysis of the BSRP disclosed herein that comprises the amino acid sequence of SEQ ID NO: 198 (pA56608_l). FIG.9C shows the tertiary structure based on in-silico analysis of the BSRP disclosed herein that comprises the amino acid sequence of SEQ ID NO: 207 (pA56621_2). FIG. 9D shows the tertiary structure based on in-silico analysis of the BSRP disclosed herein that comprises the amino acid sequence of SEQ ID NO: 260 (pA60321_l).

[0069]

[0039] FIG. 10 depicts the structure of beta sheets during the process of making a formic acid cast film.

[0070]

[0040] FIGs. 11A and 11B depict spectral data, overall fits, and component fits for aqueous (aq) and formic acid (FA) prepared samples for 6 exemplar polypeptides. The y-axis is offset for the aqueous samples for visual clarity; all data is normalized such that the maximum signal is equal to 1.

[0071]

[0041] FIG. 12 is a bar chart showing the percentages of inter- and intra- molecular beta sheets for each exemplar polypeptide and aqueous (aq) vs. formic acid (FA) prepared samples.

[0072]

[0042] FIG. 13 illustrates screen captures from video of a typical negative result from a control solution in a spinnability test.

[0073]

[0043] FIG. 14 illustrates screen captures from video of a typical positive test from a formic acid polypeptide solution.

[0074]

[0044] FIG. 15 shows typical force displacement curves from samples that drew or spun fiber during the test.

[0075] 10

[0076] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0077]

[0045] FIG. 16 is a graph showing the mass loss of droplets of polypeptide in formic acid over time.

[0078]

[0046] FIG. 17 illustrates the process of drawing a fiber and then tensile testing the fiber.

[0079]

[0047] FIG. 18 shows images of the pA55737 tensile test on texture analyzer as well as cross-sectional microscope images of fiber at break.

[0080]

[0048] FIG. 19 illustrates a drawn fiber (pA55737) captured in adhesive frame from texture analyzer: in frame (top panel), mounted for tensile testing (lower left panel), and SEM of cross-sectional area (lower center and right panels).

[0081]

[0049] FIG. 20 shows an estimated stress strain curve for a dried fiber based on the observed cross section of the broken fiber.

[0082]

[0050] FIG. 21 A shows the WAXS pattern for unstretched pA56738 in black on white and FIG.

[0083] 21B shows the WAXS pattern for unstretched pA56738 in color.

[0084]

[0051] FIG. 22 shows the WAXS pattern for unstretched pA56738 in white on black.

[0085]

[0052] FIG. 23A shows the WAXS pattern for stretched pA56738 in black on white and FIG.

[0086] 23B shows the WAXS pattern for stretched pA56738 in color.

[0087]

[0053] FIG. 24 shows the WAXS pattern for stretched pA56738 in white on black.

[0088]

[0054] FIGs. 25A and 25B depict schematics of idealized structures and scattering patterns of beta sheet crystals in fibers with varying beta strand orientation.

[0089]

[0055] FIG. 26 shows azimuthally integrated intensity profile for the unstretched pA56738 polypeptide fiber.

[0090]

[0056] FIG. 27 shows azimuthally integrated intensity profile for the stretched pA56738 polypeptide fiber.

[0091]

[0057] FIG. 28 shows radially integrated azimuthal profile plots of the T (inter-sheet) scattering feature for the unstretched and stretched pA56738 fibers.

[0092]

[0058] FIG. 29 shows radially integrated azimuthal profile plots of the (interstrand) scattering feature for the unstretched and stretched pA56738 synthetic fibers.

[0093] 11

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[0095]

[0059] FIG. 30 shows polarized Raman orientation analysis of fiber pA56738 stretched (top panel) or unstretched (bottom panel). In both panels, the solid line is the spectrum polarized parallel to the fiber (both excitation and detection) and the dotted line is the spectrum polarized perpendicular to the fiber (both excitation and detection).

[0096]

[0060] FIG. 31 shows a schematic of chains in the fibers produced based on the WAXS and polarized Raman analyses before and after drawing or stretching the fiber.

[0097]

[0061] FIG. 32A shows the WAXS pattern for stretched pA55724 fiber in black on white and FIG. 32B shows the WAXS pattern for stretched pA55724 in color.

[0098]

[0062] FIG. 33 shows the WAXS pattern for stretched pA55724 fiber in white on black.

[0099]

[0063] FIG. 34 shows the polarized Raman orientation analysis for pA55724 stretched fiber (top panel) and unstretched fiber (bottom panel). The solid and dotted lines are the same as described for FIG. 30.

[0100]

[0064] FIG. 35 shows the stress v strain curve for pA55724.

[0101]

[0065] FIG. 36 shows a graph of the TGA curve for fiber pA55724.

[0102]

[0066] FIG. 37 shows the results of DMA tests on the experimental pA55724 fibers in comparison to a commercial Dyneema® high performance filament.

[0103]

[0067] FIG. 38 shows stress v strain curve for pA56738.

[0104]

[0068] FIG. 39 shows a graph of the TGA curve for fiber pA56738.

[0105]

[0069] FIG. 40 shows the results of DMA tests on the experimental pA56738 fibers in comparison to a commercial Dyneema® high performance filament.

[0106]

[0070] FIG. 41 shows a Scanning Electron Microscopy (SEM) image of a produced fiber web.

[0107]

[0071] FIG. 42 depicts a cross section of electrospun mat with aluminum backer (solid material) and electrospun polypeptide.

[0108]

[0072] FIG. 43 shows a fiber diameter distribution based on images of the pA55737 fiber.

[0109]

[0073] FIG. 44 shows the surface pore size distribution of a nanofiber mat made of the pA55737 polypeptide.

[0110] 12

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[0112]

[0074] FIG. 45 shows a stress v strain curve for a pA56739 film.

[0113]

[0075] FIG. 46 shows polarized Raman orientation analysis of cast films from formic acid (top) and water (bottom), demonstrating random orientation of beta strands in pA56738 film for both aqueous and formic acid. The solid and dotted lines are the same as described for FIG. 30 but parallel or perpendicular to the long axis of the cast dogbone film.

[0114]

[0076] FIG. 47 shows a Scanning Electron Microscope (SEM) image of a cross section of a composite material prepared as described in Example 17. The blue arrows in the image indicate a length measurement performed by software to indicate the thickness of each layer of the composite material. The total thickness is 12.4 micron, the middle layer is 6 micron, the top layer according to the direction of this cross-sectional image is 4 micron, and the bottom layer is 2.4 micron. These values are approximate values due to minor variation in thickness along the length of the cross section.

[0115]

[0077] FIG. 48 shows images of a control, uncoated membrane before and after the assay for thermal stability, as described in Example 18. The image on the left shows the control materials prior to the testing for thermal stability. The image on the right shows the control material post thermal treatment. The yellow disc on the right represents the original sample dimensions, while the outlined clear samples on the right represent the membrane dimensions post-test.

[0116]

[0078] FIG. 49 shows an image demonstrating the thermal stability of a composite material prepared as described in Example 17 and tested as described in Example 18. The purple disc on the left represents the original sample dimensions, while the tan disk on the right is the composite material following the assay described in Example 18.

[0117]

[0079] FIG. 50 shows an image of the turbid suspension of fibrillar protein after injection into ethanol coagulant (right tube), compared to pure ethanol (left tube), as described in Example 22. This image illustrates optical texture.

[0118]

[0080] FIG. 51 shows images of Scanning Electron Micrographs at increasing magnification of a fibrillar / fiber network enmeshing aluminum oxide particles as described in Example 22.

[0119]

[0081] FIG. 52A depicts the design of the pA55743 sequence, with labeled glycine-rich peptide segments (linkers) and 8 polypeptide stretches (structural beta domains). FIG. 52B depicts the 13

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[0121] sequence analysis of linkers and structural beta domains. The tables present the glycine % and length for all linkers and structural beta domains. FIG.52C depicts the sequence of each structural beta domain with each other. FIG. 52D depicts structural beta domains modeled as separate polypeptide chains with pLDDT values depicted in grayscale. The average pLDDT of this model is 96.1. FIG. 52E depicts the 8 structural beta domains modeled as separate chains, showing D4 symmetry axes, with alternating chains colored light grey or dark grey. The coordinate origin is also displayed, with the Z axis aligned to the primary 4-fold symmetry axis, and the secondary symmetry axis aligned to the Y axis. FIG. 52F depicts the structural beta domain DSSP4 secondary structure analysis, based on a structural model. This table summarizes the DSSP4 secondary structure, with loops and strands colored in shades of gray, and the strand percentage calculated per chain.

[0122]

[0082] FIGs. 53A-53D show pA56642, a non-naturally occurring beta solenoid protein comprising a large sheet folded upon itself. FIG. 53A depicts the structural model showing strand residues (by DSSP4 secondary structure) have an average pLDDT > 60. FIG. 53B shows the structural model with DSSP4 secondary structure and spine residues shown in grayscale. The main sheet forms two planes (labeled on the left). The strands that span both planes of the sheet are labeled on the right. The negative bend residues in the strands that span both planes are also labeled on the right. FIG. 53C shows the structural model colored by DSSP4 secondary structure, spine residues and N- and C-terminal structural caps. The sequences of the N- and C-terminal structural caps and the sequences before and after of the adjacent spine residues, respectively, are presented in a table to show the low level of homology between the caps and the adjacent solenoid sections.

[0123] FIG. 53D shows the structural model showing the DSSP4 secondary structure and spine residues in grayscale. On the right, the local geometry near a spine residue is shown: yellow lines trace the angle between residue i-3, a solenoid spine residue, and residue i+5, which is 45.6°; yellow lines show the distance between residue i-3 and i+5, which is 8.8 A. On the left, distances between adjacent spine residues are labeled, showing that the spine residues are all part of a single graph with connections of 5.8 A or less between them. The calculated strand content in a 11 -residue window around the solenoid spine residues is 86%.

[0124]

[0083] FIGs. 54A-54B show pA56629, a non-naturally occurring beta antiparallel solenoid protein comprising two sheets that are in contact with each other and are both composed of 14

[0125] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0126] antiparallel hairpins. FIG. 54A depicts the structural model showing pLDDT in grayscale. The average pLDDT of strand residues (by DSSP4 secondary structure) is 72.75. FIG. 54B shows the structural model with DSSP4 secondary structure in grayscale, with labeled sheets and an antiparallel hairpin.

[0127]

[0084] FIG. 55 shows a flow chart depicting the method used to design the BSRPs that comprise beta domains with dihedral symmetry.

[0128]

[0085] FIG. 56 shows a flow chart depicting the method used to design the BSRPs that comprise solenoid structures.

[0129]

[0086] FIGs. 57A-57C depict (i) cross-sections consistent with an asymmetric membrane architecture (a denser surface region supported by a porous sublayer) and (ii) a surface view showing pore openings. FIG. 57A depicts cross-section at ~500x (scale bar 100 pm). FIG. 57B depicts cross-section at ~l,000x (scale bar 50 pm). FIG. 57C depicts surface view at ~500x (scale bar 100 pm).

[0130] DETAILED DESCRIPTION

[0131]

[0087] Disclosed herein are beta strand reservoir polypeptides (BSRPs) designed de novo that are determined to form compositions exhibiting quaternary structures having mechanical and functional properties useful in a wide range of applications from textiles to biomedical devices. The polypeptides are expressed first in a tertiary structure compatible with or advantageous for recombinant expression techniques, purified, and processed into polymers exhibiting quaternary structures. The polypeptides provide a reservoir of beta strands that may be processed such that they transition from a tertiary structure that is more reliant on intramolecular interactions to a quaternary structure that is more reliant on intermolecular interactions.

[0132]

[0088] As detailed below, the disclosure thus provides de novo designed and synthesized beta strand reservoir polypeptides that upon processing by the methods disclosed herein including dissolving in formic acid and spinning into fibers, e.g., as described in Examples 5 and 8, denature and refold to form a beta sheet crystalline phase.

[0133] 15

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[0135]

[0089] As disclosed herein in detail, the relative ease with which the beta strand reservoir polypeptides are able to be resuspended in environmentally friendly solvents, like formic acid and the lack of gelling or other premature intermolecular association of the polypeptides within the (e.g. formic acid) dope prior to spinning are unexpected and superior properties of the beta strand reservoir polypeptides disclosed herein. Notably, solubility in formic acid is not seen in naturally occurring fibroin or spidroins. Another advantage of the disclosed BSRPs is that they can be produced without lengthy and expensive regeneration processes that require use of Lithium, Calcium, or other ionic salts, in contrast to the naturally occurring fibroin or spidroin.

[0136]

[0090] The refolding to form a beta sheet crystalline phase comprises formation of 2-dimensional beta sheets, which interact amongst each other to form 3 -dimensional stacks of a plurality of 2-dimensional beta sheet polymers called beta sheet crystalline phases. In embodiments, the beta sheet crystalline phase has desired mechanical characteristics, such as, high tensile strength. In embodiments, the processing comprises denaturation of the tertiary structure of beta strand reservoir polypeptides exhibiting more intramolecular interactions than intermolecular interactions, followed by refolding to form a beta sheet crystalline phase disclosed herein comprising a quaternary structure exhibiting more intermolecular interactions than intramolecular interactions, as further detailed below.

[0137]

[0091] The following description includes information that may be useful in understanding the present disclosure. It is not an admission that any of the information provided herein is prior art or relevant to the presently embodimented disclosures, or that any publication specifically or implicitly referenced is prior art.

[0138] Definitions

[0139]

[0092] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0140]

[0093] All technical and scientific terms used herein, unless otherwise defined below, are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to techniques employed herein are intended to refer to the techniques as commonly understood in

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[0143] the art, including variations on those techniques and / or substitutions of equivalent techniques that would be apparent to one of skill in the art.

[0144]

[0094] Any ranges listed herein are intended to be inclusive of endpoints. For example, a range of 2-4 includes 2 and 4.

[0145]

[0095] Also as used herein, “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).

[0146]

[0096] In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. The term “about”, when immediately preceding a number or numeral, means that the number or numeral ranges plus or minus 10%.

[0147]

[0097] As used herein, a “non-naturally occurring polypeptide” is a polypeptide that does not occur in nature. In embodiments, a non-naturally occurring polypeptide may be designed de novo. In embodiments, a “non-naturally occurring polypeptide” is a beta strand reservoir polypeptide described herein.

[0148]

[0098] As used herein, a “non-naturally occurring polymer” is a polymer, such as a polypeptide, that does not occur in nature. In embodiments, a non-naturally occurring polymer may be designed starting with a naturally occurring polypeptide or not starting with a naturally occurring polypeptide. In embodiments, a “non-naturally occurring polymer” is a beta strand reservoir polypeptide polymer described herein.

[0149]

[0099] Unless otherwise noted, all secondary or tertiary structures referenced herein were determined based on in silico prediction and analyses using at least one or both of ColabFold and ESMFold. Unless otherwise noted, the quaternary structure of beta domains, such as, shown in FIGs. 3A-3D and 4A-4B is determined by in silico prediction and analyses.

[0150]

[0100] For polypeptides designed to have internal dihedral pseudosymmetry, an atomic model of the beta domain complex is generated using ColabFold in the following fashion: A sequence string is created, containing 8 (D4 designs) or 12 (D6 designs) copies of the beta domain sequence,

[0151] 17

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[0153] separated by a then this string is provided to ColabFold 1.5.2 (From commit 62fb2a4aa15dc7fdd8cc7d710749f907b1e8cf2f of github.com / sokrypton / ColabFold.git “AlphaFold2_batch” Notebook / mode) as input, using the following options: model-type=alphafold2_multimer_v3 rank=plddt num-relax=l use-gpu-relax=True num-recycle= 5 num-models=3 disable-unified-memory=True msa-mode=single_sequence. All other options not mentioned are left as their default values. The model considered as unique output is the top one by pLDDT.

[0154]

[0101] For polypeptides designed to have a beta solenoid structure an atomic model is generated using ColabFold and ESMFold in the following fashion: First two models are generated, one with ColabFold: A sequence string is created with the design sequence, then this string is provided to ColabFold 1.5.2 (From commit 62fb2a4aa15dc7fdd8cc7d710749f907b1e8cf2f of github.com / sokrypton / ColabFold.git “AlphaFold2_batch” Notebook / mode) as input, using the following options: model -type=alphafold2_multimer_v3 rank=plddt num-relax=l use-gpu-relax=True num-recycle=5 num-models=3 disable-unified-memory=True msa-mode=single_sequence. All other options not mentioned here are left as their default values. For ColabFold, the model considered as unique output is the top one by pLDDT. A second model is generated using ESMFold: A sequence string is created with the design sequence, then this string is provided to ESMFold (From commit c9c7d4ffec964ce10c3e11dccec6c16edaa5144 of github.com / facebookresearch / esm.git) as input, using the following options: -i <input sequence in fasta format> -o <output directory> -m <path to model> — max-tokens-per-batch=0 — cpu-offload. Once the ColabFold and ESMFold models are generated, one of them is selected as the final output model, depending on their structure and average pLDDT as follows: if only one of them exhibited a beta solenoid-type structure, that is considered the final output. If both exhibited a beta solenoidtype structure, the one with the highest pLDDT is considered the final output. If none of the two exhibited a beta solenoid-type structure, the one with the highest pLDDT is considered the final output.

[0155]

[0102] Further details are provided in “Biological structure and function emerge from scaling unsupervised learning to 250 million protein sequences” (2020, December 16) Proceedings of the National Academy of Sciences (PLANS') 118.15 (2021): e2016239118, and " ColabFold: making

[0156] 18

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[0158] protein folding accessible to all." Nature methods 19.6 (2022, May 30): 679-682, the contents of each of which is herein incorporated by reference in their entireties for all purposes.

[0159]

[0103] As used herein “sequence identity” refers to the extent to which two optimally-aligned polynucleotides or polypeptide sequences are invariant throughout a window of alignment of components, e.g. nucleotides or amino acids. An “identity fraction” for aligned segments of a test sequence and a reference sequence is the number of identical components which are shared by the two aligned sequences divided by the total number of components in the reference sequence segment, i.e. the entire reference sequence or a smaller defined part of the reference sequence. “Percent identity” is the identity fraction times 100. The extent of identity (homology) between two sequences can be ascertained using a computer program and mathematical algorithm. Percentage identity can be calculated using the alignment program Clustal Omega, available at ebi.ac.uk / Tools / msa / clustalo using default parameters. See, Sievers et al., “Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega.” (2011 October 11) Molecular systems biology 7:539.

[0160]

[0104] As used herein, a beta strand reservoir polypeptide (BSRP), also referred to as a beta strand reservoir polymer, is a non-naturally occurring polypeptide that comprises a tertiary structure with a plurality of beta strands participating in intramolecular strand-strand interactions, which can act as a source or “reservoir” of beta strands for the formation of quaternary structures mediated by a plurality of intermolecular beta strand-strand interactions. Without being bound by theory, it is thought that BSRPs fold, under standard protein expression conditions, into a tertiary structure that is comprised primarily of intramolecular beta strand-strand interactions as opposed to intermolecular beta strand-strand interactions. As described herein, under appropriate processing conditions, the BSRPs can denature and refold into a quaternary structure that is comprised primarily of intermolecular beta strand-strand interactions as compared to intramolecular beta strand-strand interactions. In embodiments, a BSRP disclosed herein has an amino acid sequence of at least about 80% identity to an amino acid sequence of any one of SEQ ID NOs: 1-84.

[0161]

[0105] As used herein, a “beta strand” refers to a stretch of amino acid residues of a polypeptide (such as a BSRP) that exhibits a propensity to form a secondary structure where the polypeptide chain is nearly linear and wherein hydrogen bonds form between carbonyl and amino groups of

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[0164] backbone atoms in adjacent beta strands to form beta sheets. Beta strands comprising a given beta sheet may be discontinuous in the primary sequence and can be separated by long sequences of amino acids. A beta strand within a BSRP may be identified using the program, DSSP (Define Secondary Structure of Proteins) - 2.2.1 application MKDSSP as follows: Run mkdssp on a 3D structural model of the BSRP obtained using the in silico methods described herein. Then isolate column 5 of the mkdssp text output, which contains the per-position 8-class secondary structure assignment in each line, convert all the E and B assignments to strand (E), and discard strands that are less than 3 amino acids long by discarding all isolated occurrences of ‘E’ or ‘EE’. Further details are provided in “Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features” Biopolymers (1983, December) 2577-2637, the contents of which are incorporated by reference in its entirety for all purposes.

[0165]

[0106] A beta strand may be identified using DSSP4 (version 4.4.10 from https: / / github.com / PDB-REDO / dssp).

[0166]

[0107] As used herein, the DSSP4 secondary structure of a protein structural model is the secondary structure assigned by DSSP4 (version 4.4.10 from https: / / github.com / PDB-REDO / dssp): Briefly, to assign helix, strand or loop secondary structure to each residue using DSSP4, DSSP4 is run on the PDB file output by the modeling program, using the “-output-format dssp” flag, like so: < DSSP4 build folder> / mkdssp <model PDB file> -output-format dssp. This command will output a per-residue table, preceded by a number of header lines. The table entries are isolated, the entries in column number 5 are inspected, and assignments made as follows: strand to residues with the “E” or “B” labels, helix to the residues with the “H” and “G” labels, and loop to the residues with the “S”, “T”, “C”, “.” or “P” labels.

[0167]

[0108] As used herein, a “beta sheet” is a secondary structural element of a polypeptide (e.g., a BSRP) comprising a plurality of beta strands, wherein adjacent beta strands interact with each other via hydrogen bonding between backbone carbonyl and amino groups, and wherein the amino acid side chains extend above and below the plane of the sheet. A beta sheet may be parallel or antiparallel. In a parallel beta sheet, the N terminus to C terminus orientation of all the beta strands forming the beta sheet is in the same direction. In an anti-parallel beta sheet, the N terminus to C terminus orientation of successive beta strands is in alternating directions. In embodiments, the

[0168] 20

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[0170] presence of beta sheets is indicated by the presence of an amide I band at around 1610-1630 cm'1(e.g., in the case of beta sheets that exhibit interm olecular interactions) or the presence of an amide I band at around 1630-1641 cm'1(e.g, in the case of beta sheets that exhibit intramolecular interactions), using infrared spectroscopy.

[0171]

[0109] As used herein, a “beta domain” comprises a stretch of amino acid residues of a BSRP having internal dihedral pseudo-symmetry, wherein the stretch of amino acid residues comprises a plurality of beta strands and one or more linkers, such that two beta strands are covalently joined by a linker. In embodiments, the beta domain comprises the following configuration: Lx-(S-L)y-Sz, wherein S is a beta strand and L is a linker, wherein x is 0 or 1, y is more than 4, and z is 0 or 1. In embodiments, y = 2, 3 or 4. In embodiments, all the linkers present in a beta domain are loops.

[0172] [HO] As used herein, a “linker” refers to a polypeptide comprising an amino acid sequence that covalently connects distinct amino acid stretches of a polypeptide (e.g, a BSRP), such as, beta strands, beta domains and / or beta solenoid motifs. The length of a linker is not limited and may be 1 or more amino acid residues long. A linker that is 6 or less amino acid residues in length is referred to herein as a loop. Typically, linkers do not exhibit any helix or strand secondary structure that is detectable by in silico analysis; however, in embodiments, they may present a secondary structure, including helices or strands, as detected by in silico analysis, such as, using DSSP.

[0173] [Hl] As used herein, the term “dihedral” refers to the point group symmetry of a polypeptide complex or internal pseudo-symmetry of a polypeptide monomer. A polypeptide complex is said to possess dihedral symmetry if the monomers within the complex are mathematically related to each other via two orthogonal axes of rotational symmetry, one of which is a two-fold symmetry axis. A polypeptide monomer is said to possess internal dihedral pseudo-symmetry if it has multiple domains with similar or identical sequence and structure that are mathematically related to one another via two orthogonal axes of rotational symmetry, one of which is a two-fold symmetry axis. It is important to note, as is known to those skilled in the art, that a given polypeptide monomer or complex generally exhibits small fluctuations in its structure, occupying an ensemble of states with slight variations around an energy minimum. And, in turn, a given polypeptide monomer or complex generally exhibits small deviations from perfect pseudo-

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[0176] symmetry or symmetry, respectively. The terms “dihedral symmetry” and “internal dihedral pseudo-symmetry” as used herein thus also refer to structures with small deviations from perfect symmetry or pseudo-symmetry, as reasonably determined by one with ordinary skill in the art. The disclosure provides beta strand reservoir polypeptides (BSRPs) that exhibit a tertiary structure with internal dihedral pseudo-symmetry. Provided herein are beta domains that are determined by in silico prediction and analyses to oligomerize into assemblies with dihedral symmetry. These beta domains may be connected to each other using linkers to form a BSRP. Such a BSRP is referred to herein as having a tertiary structure with internal dihedral pseudo-symmetry.

[0177]

[0112] As used herein, a “beta solenoid motif’ is a stretch of amino acid residues of a BSRP having a beta solenoid structure comprising two beta strands of opposing direction and without backbone hydrogen bond contacts between them, separated by a loop. An illustrative beta solenoid motif is depicted in FIG.8B. In embodiments, the BSRPs having a beta solenoid structure disclosed herein comprise a plurality of beta solenoid motifs covalently joined by linkers. In embodiments, one or more beta solenoid motifs of a beta solenoid BSRP may, on in silico analysis, present as a single extended beta strand of up to around 22 amino acids, wherein the loop residues are identified by a program, such as DSSP, as having beta strand secondary structure, despite the loop residues exhibiting an approximately 180 degree turn in the orientation of the polypeptide backbone and connecting segments of the extended beta strand that pair with neighboring strands from the two different beta sheets comprised by the solenoid. For instance, in FIG. 7B, the three beta solenoid motifs enveloped by a blue dotted rectangle are each identified as single continuous beta strands by the DSSP program because the loop, despite exhibiting an approximately 180 degree turn in the direction of the polypeptide chain and possessing a highly similar structure to the rest of the loops within the solenoid, is detected as having enough hydrogen bond pairings with backbone atoms in the neighboring loops to be designated as strand by the DSSP program.

[0178]

[0113] As used herein, a “beta sheet crystalline phase” refers to a solid crystalline material, wherein the material comprises polypeptides that form beta sheets that exhibit intermolecular interactions. Typically, the material comprises a 3-dimensional stack of a plurality of 2-dimensional beta sheets having molecular interactions between adjacent beta sheets. In embodiments, the beta sheet crystalline phase was formed by a process comprising introducing intermolecular interactions among a plurality of BSRPs disclosed herein. In embodiments, the 22

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[0180] plurality of BSRPs have the same amino acid sequence. In embodiments, the plurality of BSRPs have different amino acid sequences. In embodiments, the beta sheet crystalline phase is a film, a fiber, a filament or the like. In embodiments, the presence of crystalline material is evaluated by X-ray Diffraction (XRD), e.g., as described in Example 9. In embodiments, the presence of beta sheets that exhibit intermolecular interactions is indicated by the presence of an amide I band at around 1610-1630 cm'1using infrared spectroscopy. In embodiments, the beta sheet crystalline phase comprises a plurality of crystallites.

[0181]

[0114] As used herein, “pLDDT” stands for “predicted Local Distance Difference Test”, which is a per-residue measure of local confidence of the polypeptide structure predicted by ColabFold and ESMFold. It is scaled from 0 to 100, with higher scores indicating higher confidence. In other words, pLDDT measures confidence in the local structure, estimating how well the prediction would agree with an experimentally determined (e.g. via X-ray crystallography) structure. For instance, a pLDDT above 90 relates to the highest accuracy category, in which both the backbone and side chains are typically predicted with high accuracy.

[0182]

[0115] As used herein, the term “inter-sheet spacing” refers to the distance between adjacent beta sheets in a sample, as measured by Wide Angle X-ray Diffraction or Wide Angle X-ray Scattering (WAXS). Wide Angle X-ray Diffraction can be performed, for example, as described in Example 9.

[0183]

[0116] As used herein, the term “inter-strand spacing” refers to the distance between adjacent beta strands in the same beta sheet in a sample, as measured by Wide Angle X-ray Diffraction or Wide Angle X-ray Scattering (WAXS). Wide Angle X-ray Diffraction can be performed, for example, as described in Example 9.

[0184]

[0117] As used herein, the term “polymer” refers to a single covalently bonded organic macromolecular chain. In embodiments, the polymer is a polypeptide. A phase described herein may comprise a plurality of copies of the polymer.

[0185]

[0118] As used herein, a “polymer block motif’ refers to a recognizable pattern, structural arrangement, or functional unit in a polymer, analogous or equivalent to motifs in protein science, that may impart distinct physical, chemical, or biological characteristics to each block. For instance, a beta-strand motif may drive certain self-assembly or mechanical properties, whereas a 23

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[0187] non-b eta- strand motif (e.g., a helical region) may contribute different mechanical or functional attributes. A polymer block may comprise a motif, or plurality of motifs, which may or may not take the form of protein structural elements as described elsewhere in the disclosure here.

[0188]

[0119] As used herein, a “positive bend residue” refers to an amino acid residue characterized by DSSP4 as being bent (“S” under the “ dssp struct summary.bend” column in the DSSP4 mmCIF-formatted output) and having with positive chirality (“+” under the “ dssp struct summary. chirality” column in the DSSP4 mmCIF-formatted output).

[0189]

[0120] As used herein, a “glycine solenoid spine” is a network of residues that can be found in a given beta solenoid model such that it has the following characteristics:

[0190] i. The network edges are pairwise alpha carbon distances of 5.8 A or less ii. The network has at least 3 residues

[0191] iii. All residues are positive bend residues

[0192] iv. None of the residues are the first one, second one or third one in the sequence

[0193] v. If N is the length of the sequence, none of the residues are in the N-4, N-3,

[0194] N-2, N-l orN sequence position

[0195] vi. For each residue, whose position in the sequence is i, the C alpha carbon of residue i-3 and the C alpha carbon of i+5 is between 5 A and 10 A.

[0196] vii. For each residue, whose position in the sequence is i, the residues i-2, i-1, i+1 and i+2 are glycine.

[0197] viii. For each residue, whose position in the sequence is i, the angle between the vector from the C alpha carbon of said spine residue, to the C alpha carbon of the i-3 residue and the vector from the C alpha carbon of said spine residue, to the C alpha carbon of the i+5 residue, is between 20° and 40°. ix. None of the residues are consecutive in sequence

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[0200] x. 50% of all residues within a -7 and +8 residue window of spine residues are in beta strand conformation (by DSSP4 secondary structure analysis of the structure model)

[0201]

[0121] As used herein, given a beta solenoid structure model that has one or more solenoid spines or glycine solenoid spines, a “beta solenoid hairpin” refers to a polypeptide segment that is 16 residues long and has a spine residue at position 8.

[0202]

[0122] As used herein, a “droplet drawing test” (or interchangeably referred to herein as “spinnability test”) refers to a method for assessing fiber formation or spinnability for submilligram quantities of protein or polymer, for example, in the presence of volatile and / or corrosive organic solvents. In embodiments, the test comprises the following: A droplet (e.g., of 5 ul or less) of the solution may be placed on a flat surface, e.g., a fluorinated film surface. The droplet may be in a convective air stream, e.g., allowing the solution concentration to increase over time. A rod may be brought into contact with the surface of the droplet. The rod may be withdrawn from the droplet. Fiber formation between the rod and the droplet may be observed, e.g., optically or via a force transducer. This process may be repeated, e.g., for about 15 minutes or until the liquid has evaporated or an observed fiber is formed. The presence or absence of a fiber may be noted.

[0203] Beta strand reservoir polypeptides

[0204]

[0123] The disclosure provides non-naturally occurring polypeptides that have been designed de novo to be rich in beta strands and are referred to herein as “beta strand reservoir polypeptides”. The beta strand reservoir polypeptides (BSRPs) disclosed herein may be used to create compositions with desired structures and functions. As expressed, the beta strand reservoir polypeptides are determined based on in silico analysis to exhibit a tertiary structure comprising more intramolecular interactions than intermolecular interactions. Illustrative tertiary structure types include: (i) a structure with dihedral pseudo symmetry, (ii) a beta solenoid structure comprising beta solenoid motifs, and (iii) an anti-parallel beta sheet structure.

[0205]

[0124] In embodiments, the beta strand reservoir polypeptide disclosed herein comprises an amino acid sequence that is at least 75% (for instance, about 80% about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or

[0206] 25

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[0208] 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of SEQ ID NOs: 1-84.

[0209]

[0125] BSRPs may be processed into higher order structures. Processing of a BSRP using the methods disclosed herein results in the denaturation of their determined tertiary structure followed by refolding to form a beta sheet crystalline phase comprising a quaternary structure having more intermolecular interactions as compared to the beta strand reservoir polypeptide, as shown in FIG. ID. Without being bound by theory, it is thought that the presence of a large number of beta strands capable of promiscuous inter-strand interactions enables the beta strand reservoir polypeptides to readily transition from a tertiary structure that is more reliant on intramolecular interactions to a quaternary structure that is more reliant on intermolecular interactions.

[0210]

[0126] The proportion of beta sheets that exhibit intermolecular interactions vs. intramolecular interactions may be analyzed using infrared spectroscopy. In embodiments, the presence of an amide I band between 1610 cm'1and 1630 cm'1indicates the presence of intermolecular interactions between beta sheets. In embodiments, the amide I band is centered at or around 1624 cm'1. On the other hand, the presence of an amide I band between 1630 cm'1and 1640 cm'1indicates the presence of intramolecular interactions between beta sheets. In embodiments, the amide I band is centered at or around 1630 cm'1. Further details are provided in Hu et al., Macromolecules, Vol. 39, No. 18, 2006, the contents of which are incorporated herein by reference in its entirety. The transition from a tertiary structure that is more reliant on intramolecular interactions to a quaternary structure that is more reliant on intermolecular interactions may be assayed by observing: (i) an increase in the amide I band between 1610 cm'1and 1630 cm'1, (ii) a decrease in the amide I band between 1630 cm'1and 1640 cm'1, or (iii) both.

[0211]

[0127] The BSRPs disclosed herein therefore have the superior property of being able to form a beta sheet crystalline phase, often with desired mechanical characteristics.

[0212]

[0128] The molecular weight of the beta strand reservoir polypeptide may be in the range of about 50 kDa to about 200 kDa, for instance, about 50 kDa, about 55 kDa, about 60 kDa, about 65 kDa, about 70 kDa, about 75 kDa, about 80 kDa, about 85 kDa, about 90 kDa, about 95 kDa, about 100 kDa, about 105 kDa, about 110 kDa, about 115 kDa, about 120 kDa, about 125 kDa, about 130 kDa, about 135 kDa, about 140 kDa, about 145 kDa, about 150 kDa, about 155 kDa, about 160

[0213] 26

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[0215] kDa, about 165 kDa, about 170 kDa, about 175 kDa, about 180 kDa, about 185 kDa, about 190 kDa, about 195 kDa, or about 200 kDa, inclusive of all values and subranges that lie therebetween.

[0216]

[0129] In embodiments, the molecular weight of the beta strand reservoir polypeptide is more than 200 kDa, for instance, in the range of about 200 kDa to about 300 kDa, in the range of about 300 kDa to about 400 kDa, in the range of about 400 kDa to about 500 kDa, in the range of about 500 kDa to about 600 kDa, in the range of about 600 kDa to about 700 kDa, in the range of about 700 kDa to about 800 kDa, in the range of about 800 kDa to about 900 kDa, or in the range of about 900 kDa to about 1000 kDa. In embodiments, the molecular weight of the BSRP polypeptide or a polypeptide derived therefrom is more than 1,000 kDa, 10,000 kDa, 100,000 kDa, IxlO6kDa, IxlO7kDa, IxlO8kDa, or IxlO9kDa.

[0217]

[0130] In embodiments, high molecular weights may be generated by concatenation. Suitable concatenation methods include the concatenation of the DNA sequences encoding two BSRPs with the DNA sequence encoding a linker in between, so they are expressed as a single polypeptide (e.g., SEQ ID NO: 84, which is a concatenation of SEQ ID NO: 82, two copies of SEQ ID NO: 166 and SEQ ID NO: 83), disulfide bonding, enzyme catalyzed isopeptide bond formation, use of paired terminal binding moieties, split inteins and autocatalytic isopeptide bond formation, such as SpyTag / SpyCatcher conjugation. Accordingly, in embodiments, a BSRP described herein has an amino acid sequence comprising SEQ ID NO: 82, a linker (e.g., wherein the linker comprises two copies of SEQ ID NO: 116), and SEQ ID NO: 83, or an amino acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% identity thereto. In embodiments, a beta sheet crystalline phase described herein comprises a polypeptide having an amino acid sequence comprising SEQ ID NO: 82, a linker (e.g., wherein the linker comprises two copies of SEQ ID NO: 116), and SEQ ID NO: 83, or an amino acid sequence having at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% identity thereto. Further details are provided in U. S. Patent No. 9,547,003, U. S. Patent No. 11,059,867 and U. S. Publication No. 20220135628, Keeble, Anthony H., et al. " Approaching infinite affinity through engineering of peptide-protein interaction." Proceedings of the National Academy of Sciences 116.52 (2019): 26523-26533 and Zakeri, Bijan, et al. " Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin." Proceedings of the National Academy of Sciences 109.12 (2012): E690-E69, the contents of each of which are incorporated herein by reference in its entirety for all purposes.

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[0220]

[0131] The beta strand reservoir polypeptides disclosed herein may be modified to comprise one or more markers, labels, or tags. For example, in embodiments, a beta strand reservoir polypeptide disclosed herein may be labeled with a label that will allow its detection, e.g., a radiolabel, a fluorescent agent, biotin, a peptide, an enzyme fragment, or the like. In embodiments, a beta strand reservoir polypeptide disclosed herein may comprise an affinity tag, e.g., a His-tag, a FLAG tag, a GST-tag, a Strep-tag, a biotin-tag, an immunoglobulin binding domain, e.g., an IgG binding domain, a calmodulin binding peptide, and the like. In embodiments, a beta strand reservoir polypeptide disclosed herein comprises an amino acid sequence of any one or more of SEQ ID NOs: 159-173, 1144 and 1145.

[0221]

[0132] In embodiments, the beta strand reservoir polypeptides disclosed herein comprise an amino acid sequence that has less than 80% (for instance, about 75%, about 70%, about 65%, about 60%, about 55%, about 50%, about 45%, about 40%, about 35%, about 30%, about 25% or less) of alanine and glycine. In embodiments, the beta strand reservoir polypeptides disclosed herein comprise an amino acid sequence that has less than 50% of alanine and glycine. In embodiments, the beta strand reservoir polypeptides disclosed herein comprise an amino acid sequence that has less than 35% of alanine and glycine. Unless specified otherwise, the percent of alanine and glycine in a polypeptide refers to the percent of the total number of amino acids in the polypeptide that are alanine or glycine.

[0222]

[0133] The beta strand reservoir polypeptides disclosed herein are determined based on in silico analysis to form stable tertiary structures that facilitate their expression in host cells and purification therefrom. Two kinds of tertiary structures that are determined based on in silico analysis to be formed by the beta strand reservoir polypeptides are illustrated here, in particular: tertiary structures with internal dihedral pseudo-symmetry and beta solenoid structures. In embodiments, the beta strand reservoir polypeptides that are determined based on in silico analysis to form a tertiary structure with internal dihedral pseudo-symmetry or a beta solenoid structure share the ability to denature and refold to form a beta sheet crystalline phase under suitable conditions. The structures of the BSRPs are described further below.

[0223]

[0134] In embodiments, a BSRP disclosed herein is encoded by any one of the nucleotide sequences of SEQ ID NO: 2764-2847, or a sequence with at least 70%, at least 80%, at least 90%,

[0224] 28

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[0226] at least 95%, or at least 99% identity thereto. In embodiments, a beta sheet crystalline phase comprises a polypeptide encoded by any one of the nucleotide sequences of SEQ ID NO: 2764-2847, or a sequence with at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% identity thereto.

[0227] I. Beta strand reservoir polypeptides with a Tertiary Structure Having Internal Dihedral Pseudo-symmetry

[0228]

[0135] The disclosure provides non-naturally occurring, beta strand reservoir polypeptides that are determined to exhibit a tertiary structure with internal dihedral pseudo-symmetry based on in silico structural modeling using ColabFold. In embodiments, the polypeptides disclosed herein comprise four or more beta domains and a linker between each of the beta domains. In embodiments, the polypeptides disclosed herein comprising four or more beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure with internal dihedral pseudosymmetry. The kind of internal dihedral pseudo-symmetry is not limited and may be D2, D3, D4, D5, De, D7, Ds, D9, Dio, Du, or D12.

[0229]

[0136] Each of the beta domains in a BSRP with a tertiary structure having internal dihedral pseudo-symmetry may comprise five beta strands separated by loops. In embodiments, the five beta strands fold to form two beta sheets (Bx and By), wherein Bx comprises three beta strands and By comprises two beta strands. In other words, in embodiments, the beta domains of the as-expressed BSRPs with internal dihedral pseudo-symmetry exhibit a beta sheet secondary structure.

[0230]

[0137] The disclosure provides beta domains that are determined via in silico analysis to oligomerize into assemblies with dihedral symmetry. An exemplary beta domain oligomer is shown in FIGs. 3A-3D and 4A-4B. In embodiments, the beta domains that oligomerize into assemblies with dihedral symmetry may be connected to each other using linkers (such as, linkers comprising an amino acid sequence having at least 80% identity to any one or more of SEQ ID NOs: 122-158, 175-194, 1143 and 1775-2063) to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into assemblies with dihedral symmetry may be connected to each other using linkers (such as, linkers comprising an amino acid sequence of any one or more of SEQ ID NOs: 122-158, 175-194, 1143 and 1775-2063) to form a beta strand

[0231] 29

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[0233] reservoir polypeptide exhibiting a tertiary structure with internal dihedral pseudo-symmetry matching the symmetry of the unlinked beta domains on their own.

[0234]

[0138] In embodiments, the polypeptides disclosed herein comprise a plurality of beta domains and a linker between each of the beta domains. In embodiments, the beta strand reservoir polypeptides comprising a plurality of beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure having internal Dn dihedral pseudo-symmetry, wherein n is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12. In embodiments, the disclosure provides beta domains that oligomerize into quaternary structures with Dn dihedral symmetry. In embodiments, the beta domains that oligomerize into quaternary structures with Dn dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into quaternary structures with Dn dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide exhibiting a tertiary structure with internal Dn dihedral pseudo-symmetry.

[0235]

[0139] In embodiments, the BSRPs disclosed herein comprise an amino acid sequence that is at least 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to any one of SEQ ID NOs: 2, 3, 46-54, and 80-84.

[0236]

[0140] In embodiments, the BSRPs comprise four beta domains and a linker between each of the beta domains. In embodiments, the BSRPs comprising four beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure having internal D2 dihedral pseudo-symmetry. In embodiments, the disclosure provides beta domains that oligomerize into tetramers with D2 dihedral symmetry. In embodiments, the beta domains that oligomerize into tetramers with D2 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into tetramers with D2 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide exhibiting a tertiary structure with internal D2 dihedral pseudo-symmetry.

[0237]

[0141] In embodiments, the BSRPs comprise six beta domains and a linker between each of the beta domains. In embodiments, the BSRPs comprising six beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure having internal D3 dihedral 30

[0238] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0239] pseudo-symmetry. In embodiments, the disclosure provides beta domains that oligomerize into hexamers with D3 dihedral symmetry. In embodiments, the beta domains that oligomerize into hexamers with D3 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into hexamers with D3 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide exhibiting a tertiary structure with internal D3 dihedral pseudo-symmetry.

[0240]

[0142] In embodiments, the BSRPs disclosed herein comprise eight beta domains and a linker between each of the beta domains. In embodiments, the BSRPs disclosed herein comprising eight beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure having internal D4 dihedral pseudo-symmetry. In embodiments, the disclosure provides beta domains that oligomerize into octamers with D4 dihedral symmetry. In embodiments, the beta domains that oligomerize into octamers with D4 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into octamers with D4 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide exhibiting a tertiary structure with internal D4 dihedral pseudo-symmetry.

[0241]

[0143] In embodiments, the BSRPs disclosed herein determined to exhibit a tertiary structure with internal D4 dihedral pseudo-symmetry comprise an amino acid sequence that is at least 50% (for instance, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to any one of SEQ ID NOs: 3, 46-54, and 80-83.

[0242]

[0144] In embodiments, the BSRPs comprise ten beta domains and a linker between each of the beta domains. In embodiments, the BSRPs comprising ten beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure having internal D5 dihedral pseudo-symmetry. In embodiments, the disclosure provides beta domains that oligomerize into decamers with D5 dihedral symmetry. In embodiments, the beta domains that oligomerize into decamers with D5 dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into decamers

[0243] 31

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[0245] with Ds dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide exhibiting a tertiary structure with internal Ds dihedral pseudo-symmetry.

[0246]

[0145] In embodiments, the BSRPs disclosed herein comprise 12 beta domains and a linker between each of the beta domains. In embodiments, the BSRPs disclosed herein comprising 12 beta domains and a linker between each of the beta domains are determined to fold into a tertiary structure having internal De dihedral pseudo-symmetry. In embodiments, the disclosure provides beta domains that oligomerize into dodecamers with De dihedral symmetry. In embodiments, the beta domains that oligomerize into dodecamers with De dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide. In embodiments, the beta domains that oligomerize into dodecamers with De dihedral symmetry may be connected to each other using linkers to form a beta strand reservoir polypeptide exhibiting a tertiary structure with internal De dihedral pseudo-symmetry.

[0247]

[0146] In embodiments, the beta strand reservoir polypeptides determined to exhibit a tertiary structure with internal De dihedral pseudo-symmetry comprise an amino acid sequence that is at least 50% (for instance, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to any one of SEQ ID NO: 2.

[0248]

[0147] In embodiments, one or more of the beta domains in a beta strand reservoir polypeptide disclosed herein comprises or consists of about 40 amino acid residues to about 100 amino acid residues, for example, about 40 amino acids, about 45 amino acids, about 50 amino acids, about 55 amino acids, about 60 amino acids, about 65 amino acids, about 70 amino acids, about 75 amino acids, about 80 amino acids, about 85 amino acids, about 90 amino acids, about 95 amino acids or about 100 amino acids, inclusive of all values and subranges that lie therebetween. In embodiments, one or more of the beta domains comprises or consists of about 55 amino acids.

[0249]

[0148] In embodiments, each of the beta domains in a beta strand reservoir polypeptide disclosed herein comprises or consists of about 40 amino acid residues to about 100 amino acid residues, for example, about 40 amino acids, about 45 amino acids, about 50 amino acids, about 55 amino acids, about 60 amino acids, about 65 amino acids, about 70 amino acids, about 75 amino acids, about 32

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[0251] 80 amino acids, about 85 amino acids, about 90 amino acids, about 95 amino acids or about 100 amino acids, inclusive of all values and subranges that lie therebetween. In embodiments, each of the beta domains comprises or consists of about 55 amino acids.

[0252]

[0149] The disclosure provides BSRPs comprising an even number of four or more polypeptide stretches referred to herein as “structural beta domains”, which have the same sequence length, are 40 to 100 amino acids in length, are all 75% identical or more to each other in terms of sequence, their sequence is less than 15% glycine, and are connected by glycine-rich (>30% glycine) peptide segments of 10 or more residues in length, also referred to herein as “linkers”.

[0253]

[0150] In embodiments, when modeled together as separate polypeptide chains, structural beta domains form a single complex with dihedral symmetry Dn, where n = # structural beta domains / 2, the pLDDT average over all modeled residues is above 70, and most residues (>50%) are part of a beta strand, wherein the beta strand is identified by DSSP4 secondary structure analysis of the model.

[0254]

[0151] In embodiments, when modeled together as separate polypeptide chains, structural beta domains form a single complex with cyclic symmetry Cn, where n = # structural beta domains, the pLDDT average over all modeled residues is above 70, and most residues (>50%) are part of a beta strand, wherein the beta strand is identified by DSSP4 secondary structure analysis of the model.

[0255]

[0152] Without being bound by theory, it is thought that upon spinning, the residues within the structural beta domains form a beta sheet crystalline phase due to their propensity to form beta sheets. Furthermore, the fact that their sequence is repeated or highly homologous to other sequences within the BSRP contributes to the formation of the beta sheet crystalline phase without the need of different peptide chains being perfectly aligned.

[0256]

[0153] Without being bound by theory, it is thought that the linkers, with their high glycine content and length of 10 or more residues, impart elasticity to the spun fiber.

[0257]

[0154] Without being bound by theory, it is thought that the lower number of glycines (<15%) in the structural beta domains aids its folding into a discrete conformation that aids expression and processing.

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[0260]

[0155] Without being bound by theory, it is thought that the symmetric structure that the structural beta domains form when modeled as separate chains is homologous (or similar) to one formed by the beta domains within the dihedral BSRP when expressed and dissolved in aqueous buffer. Without being bound by theory, it is thought that this conformation aids the expression of the protein and maintains the polypeptide chain in a compact soluble state that aids processing.

[0261]

[0156] As an exemplary embodiment, SEQ ID NO: 3 (pA55743) contains 8 structural beta domains, which all have 100% sequence identity amongst each other, are 55 residues in length and are 9.1% glycine (FIGs. 52A-52C). The linkers are 10 or more amino acids or long and comprise more than 30% glycine. Modeling (as described earlier for dihedral BSRPs) of the structural beta domains detected in the sequence further verifies that: 65% of residues in each structural beta domain, modeled as a complex of separate chains, are part of a beta strand identified by DSSP4 secondary structure analysis of the model (FIGs.52D-52F). The average pLDDT over all modeled residues was calculated by inspecting the B-factor column in the output model, summing said column, and dividing by the total number of modeled residues. Inspecting the same model, it was determined if the structural beta domains are arranged in a single complex, with Dn symmetry or pseudosymmetry (in the case chains have non-identical sequences), where n = number of structural beta domains / 2, or Cn symmetry or pseudosymmetry (in the case chains have non-identical sequences), where n = number of structural beta domains.

[0262] (a) Beta Strands

[0263]

[0157] As illustrated in FIG. 1A, beta strands, along with linkers, form the building blocks of the beta strand reservoir polypeptides disclosed herein. The ability of beta strands to form inter-strand, backbone hydrogen bonds underlie the utility of beta strand reservoir polypeptides as components of a beta sheet crystalline phase. In other words, beta strands facilitate the formation of the tertiary structure of a beta strand reservoir polypeptide, as well as facilitate the formation of quaternary structures in a beta sheet crystalline phase.

[0264]

[0158] In embodiments, one or more of the beta strands in a beta strand reservoir polypeptide (BSRP), wherein the BSRP has a tertiary structure with internal dihedral pseudo-symmetry, comprises about 3 to about 10 amino acid residues, for instance, 3 amino acid residues, 4 amino 34

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[0266] acid residues, 5 amino acid residues, 6 amino acid residues, 7 amino acid residues, 8 amino acid residues, 9 amino acid residues or 10 amino acid residues. In embodiments, the beta strand comprises less than 50% glycine residues, for instance, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, or about 0%, inclusive of all values and subranges that lie therebetween. In embodiments, the beta strand comprises less than 50% alanine residues, for instance, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, or about 0%, inclusive of all values and subranges that lie therebetween. In embodiments, the beta strand comprises less than 50% glycine residues and / or less than 50% alanine residues. In embodiments, the beta strand comprises less than 50% glycine residues and less than 50% alanine residues. In embodiments, the beta strand comprises less than 50% glycine residues or less than 50% alanine residues.

[0267]

[0159] In embodiments, one or more of the beta strands in a BSRP having a tertiary structure with internal dihedral pseudo-symmetry disclosed herein comprises an amino acid sequence that is at least 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of SEQ ID NOs: 568-596. In embodiments, each of the beta strands in a BSRP having a tertiary structure with internal dihedral pseudo-symmetry disclosed herein comprises an amino acid sequence that is at least 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of SEQ ID NOs: 568-596.

[0268]

[0160] In embodiments, the beta strand reservoir polypeptide determined to exhibit a tertiary structure having internal dihedral pseudo-symmetry is engineered such that the interaction between the N-terminus of one polypeptide and the C-terminus of another polypeptide is prevented. For instance, in embodiments, the beta strand reservoir polypeptide is a circularized polypeptide. As used herein, a “circularized polypeptide” is a polypeptide that is engineered such that the beta strands that are at the edges of the beta sheet or sheets of one individual polypeptide are not available to interact with the edge beta strands of beta sheets from another polypeptide. As used herein, as “edge strand” is a beta strand that is on either end of a beta sheet. Typically, an edge 35

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[0270] beta strand is a beta strand at or near the N-terminus or the C-terminus of the BSRP. In embodiments, however, the N- or C-terminal beta strand may fold back into the beta sheet, exposing a different beta strand as the edge strand. Circularized polypeptides may be generated via a variety of different computational design methods. Further details are provided in Sheffler et al., Fast and versatile sequence-independent protein docking for nanomaterials design using RPXDock, PLOS Computational Biology, 2023 May 22;19(5):el010680 and Watson et al., De novo design of protein structure and function with VN diffusion, Nature, 2023 Aug; 620(7976): 1089-1100, the contents of each of which are incorporated herein by reference in their entireties for all purposes.

[0271]

[0161] Without being bound by theory, it is thought that circularization reduces or prevents interactions between the beta strands that are at the edges of the beta sheet or sheets of a polypeptide with the beta strands at the edges of the beta sheet or sheets of another polypeptide, thereby reducing the chances of formation of intermolecular polypeptide aggregates that can be problematic for expression and / or purification.

[0272] (b) Beta Domains

[0273]

[0162] As illustrated in FIG. 1A, beta domains may multimerize and form homo-oligomers. In an exemplary embodiment, 8 beta domains are determined to come together to form an octamer based on in silico analyses. As noted earlier, the beta domains that oligomerize may be linked or connected via linkers to form a beta strand reservoir polypeptide.

[0274]

[0163] In embodiments, the beta domains comprise alternating beta strands and linkers. For instance, in embodiments, one or more of the beta domains in a beta strand reservoir polypeptide with dihedral pseudo-symmetry comprises a configuration of Lx-(S-L)y-Sz, wherein S is a beta strand and L is a linker, wherein x is 0 or 1, y is any one of 4 through 12, and z is 0 or 1. In embodiments, each of the beta domains comprises a configuration of Lx-(S-L)y-Sz, wherein S is a beta strand and L is a linker, wherein x is 0 or 1, y is any one of 4 through 12, and z is 0 or 1. In embodiments, each of the beta domains comprises a configuration of Lx-(S-L)y-Sz, wherein x=l, y=4 and z=l. In embodiments, at least 30% (for instance, about 20%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about

[0275] 36

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[0277] 85%, about 90%, about 95%, or about 100%, inclusive of all values and subranges that lie therebetween) of the amino acid residues of each of the beta domains are part of a beta strand.

[0278]

[0164] In embodiments, one or more of the beta domains in a beta strand reservoir polypeptide that is determined to have a tertiary structure with internal dihedral pseudo-symmetry exhibits an average pLDDT of over 70, for instance over 75, over 80, over 85, over 90, or over 95. In embodiments, each of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT of over 70, for instance over 75, over 80, over 85, over 90, or over 95. In embodiments, one or more of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT that lies in the range of about 50 to about 100, for example about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, or about 95. In embodiments, each of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT that lies in the range of about 50 to about 100, for example about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, or about 95. In embodiments, one or more of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT that lies in the range of about 50 to about 70. In embodiments, each of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT that lies in the range of about 50 to about 70. In embodiments, one or more of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT that lies in the range of about 70 to about 90. In embodiments, each of the beta domains in a beta strand reservoir polypeptide disclosed herein exhibits an average pLDDT that lies in the range of about 70 to 100.

[0279]

[0165] In embodiments, one or more of the beta domains in a beta strand reservoir polypeptide disclosed herein comprises an amino acid sequence that is at least 75% (for instance, about 80% about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of SEQ ID NOs: 91-96. In embodiments, each of the beta domains in a beta strand reservoir polypeptide disclosed herein comprises an amino acid sequence that is at least 75% (for instance, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of SEQ ID NOS: 91-96.

[0280] 37

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[0282]

[0166] In embodiments, the beta domains do not comprise one or more of the following: (i) an amino acid sequence of (GA)nX, wherein X is any amino acid and n>5, (ii) an amino acid sequence of An, wherein n >5, and (iii) an amino acid sequence of (AG)n, wherein n >5. In embodiments, the beta domains do not comprise the following: (i) an amino acid sequence of (GA)nX, wherein X is any amino acid and n>5, (ii) an amino acid sequence of An, wherein n >5, and (iii) an amino acid sequence of (AG)n, wherein n >5.

[0283]

[0167] Table 1 lists BSRPs that are determined to exhibit a tertiary structure with internal dihedral pseudo-symmetry disclosed herein along with their respective beta domains and beta strands. The SEQ ID NO. in the first column applies to the full sequence listed in the fourth column of the table. In embodiments, the present disclosure provides an amino acid sequence of Table 1 having an initial methionine. In embodiments, the present disclosure provides an amino acid sequence of Table 1 that does not comprise an initial methionine.

[0284] 38

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[0286] Table 1:

[0287] SEQ Structure Name Full Sequence Beta Domains (separated by a Beta strands (separated by a ID of beta dash dash

[0288] NO: domains

[0289] in silico

[0290] 2 Dihedral - pA55737 TTETTPAVSVKDGETKALKVIGDKSW TTETTPAVSVKDGETKALKVIG TET-TKALKVI-KSWASRK- D6 ASRKGDTVTLTVYNAKPTDKVSVTIET DKSWASRKGDTVTLTVYNAK TVTLTVY-KVSVTIE-TET- VGQGGYGGLGGQGAGRGAGTTETTPA PTDKVSVTIETV- TKALKVI-KSWASRK- VSVKDGETKALKVIGDKSVVASRKGDT TTETTPAVSVKDGETKALKVIG TVTLTVY-KVSVTIE-TET- VTLTVYNAKPTDKVSVTIETVGGAGQG DKSWASRKGDTVTLTVYNAK TKALKVI-KSWASRK- GYGGLGSQGAGRGGYGGQGATTETTP PTDKVSVTIETV- TVTLTVY-KVSVTIE-TET-TKA- AVSVKDGETKALKVIGDKSWASRKGD TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- TVTLTVYNAKPTDKVSVTIETVGGAGQ DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKALKVI- GGYGGLGSQGAGRGGYGGQGATTETT PTDKVSVTIETV- KSWASRK-TVTLTVY- PAVSVKDGETKALKVIGDKSWASRKG TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKA- DTVTLTVYNAKPTDKVSVTIETVGGAG DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- QGGFGGLGGQGAGTTETTPAVSVKDGE PTDKVSVTIETV- KVSVTIE-TET-TKA- TKALKVIGDKSWASRKGDTVTLTVYN TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- AKPTDKVSVTIETVGGAGQGGYGGLGG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- QGAGRGAGTTETTPAVSVKDGETKALK PTDKVSVTIETV- KSWASRK-TVTLTVY- VIGDKSWASRKGDTVTLTVYNAKPTD TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- KVSVTIETVGGAGQGGFGGLGGQGAGT DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- TETTPAVSVKDGETKALKVIGDKSWA PTDKVSVTIETV- KVSVTIE-TET-TKA- SRKGDTVTLTVYNAKPTDKVSVTIETV TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- GGAGQGGYGGLGGQGAGRGAGTTETT DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- PAVSVKDGETKALKVIGDKSWASRKG PTDKVSVTIETV- KSWASRK-TVTLTVY- DTVTLTVYNAKPTDKVSVTIETVGQGG TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- YGGLGGQGAGRGAGTTETTPAVSVKD DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- GETKALKVIGDKSWASRKGDTVTLTV PTDKVSVTIETV- KVSVTIE YNAKPTDKVSVTIETVGQGGYGGLGGQ TTETTPAVSVKDGETKALKVIG GAGRGAGTTETTPAVSVKDGETKALKV DKSWASRKGDTVTLTVYNAK IGDKSWASRKGDTVTLTVYNAKPTDK PTDKVSVTIETV- VSVTIETVGGAGQGGYGGLGGQGAGQ TTETTPAVSVKDGETKALKVIG GAGTTETTPAVSVKDGETKALKVIGDK DKSWASRKGDTVTLTVYNAK SWASRKGDTVTLTVYNAKPTDKVSVT PTDKVSVTIETV-

[0291]

[0292] IETVGGAGQGGYGGLGGQGAGRGAGT TTETTPAVSVKDGETKALKVIG

[0293] 39

[0294] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0295] TETTPAVSVKDGETKALKVIGDKSWA DKSWASRKGDTVTLTVYNAK SRKGDTVTLTVYNAKPTDKVSVTIETV PTDKVSVTIETV- TTETTPAVSVKDGETKALKVIG DKSWASRKGDTVTLTVYNAK PTDKVSVTIETV

[0296] 3 Dihedral - pA55743 ALKYTIE VTESGGKAEYVWKDNTGAI ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- D4 LKAQKISNGDEVTVTTASGLKITVKAS NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- AGGAGQGGYGGLGSQGAGRGGYGGQ SGLKITVKASA- LKYTIEVTES-KAEYVWK- GAALKYTIEVTESGGKAEYVWKDNT ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- GAILKAQKISNGDEVTVTTASGLKITV NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- KASAGGAAQGGQGLGGQGALKYTIEV SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- TESGGKAEYVWKDNTGAILKAQKIS ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- NGDEVTVTTASGLKITVKASAGGAGQ NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- GGYGGLGGQGAGRGAGALKYTIEVTE SGLKITVKASA- LKYTIEVTES-KAEYVWK- SGGKAEYVWKDNTGAILKAQKISNG ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- DEVTVTTASGLKITVKASAGGAAQGG NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- QGLGGQGALKYTIEVTESGGKAEYW SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- VKDNTGAILKAQKISNGDEVTVTTASG ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- LKITVKASAGQGGYGGLGGQGAGRG NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- AGALKYTIEVTESGGKAEYVWKDNT SGLKITVKASA- LKYTIEVTES-KAEYVWK- GAILKAQKISNGDEVTVTTASGLKITV ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS KASAGGAGQGGFGGLGGQGAGALKY NTGAILKAQKISNGDEVTVTTA TIEVTESGGKAEYVWKDNTGAILKAQ SGLKITVKASA- KISNGDEVTVTTASGLKITVKASAGGA ALKYTIE VTESGGKAEYVWKD GQGGYGGLGGQGAGQGAGALKYTIE NTGAILKAQKISNGDEVTVTTA VTESGGKAEYVWKDNTGAILKAQKI SGLKITVKASA- SNGDEVTVTTASGLKITVKASA ALKYTIE VTESGGKAEYVWKD NTGAILKAQKISNGDEVTVTTA SGLKITVKASA TTETTPAVSVKDGETKALKVIGDKSW TTETTPAVSVKDGETKALKVIG TET-TKALKVI-KSWASRK- ASRKGDTVTLTVYNAKPTDKVSVTIET DKSWASRKGDTVTLTVYNAK TVTLTVY-KVSVTIE-TET- VGGAGQGGFGGLGGQGAGTTETTPAV PTDKVSVTIETV- TKALKVI-KSWASRK- SVKDGETKALKVIGDKSWASRKGDT TTETTPAVSVKDGETKALKVIG TVTLTVY-KVSVTIE-TET- VTLTVYNAKPTDKVSVTIETVGGAGQ DKSWASRKGDTVTLTVYNAK TKALKVI-KSWASRK- Dihedral - GGFGGLGGQGAGTTETTPAVSVKDGE PTDKVSVTIETV- TVTLTVY-KVSVTIE-TET-TKA- 46 D4 pA56733 TKALKVIGDKSWASRKGDTVTLTVY TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY-

[0297]

[0298] NAKPTDKVSVTIETVGGAGQGGYGGL DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKALKVI- 40

[0299] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0300] GGQGAGRGAGTTETTPAVSVKDGETK PTDKVSVTIETV- KSWASRK-TVTLTVY- ALKVIGDKSVVASRKGDTVTLTVYNA TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKA- KPTDKVSVTIETVGGAGQGGYGGLGS DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- QGAGRGATTETTPAVSVKDGETKALK PTDKVSVTIETV- KVSVTIE-TET-TKA- VIGDKSWASRKGDTVTLTVYNAKPT TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- DKVSVTIETVGGAGQGGYGGLGSQGA DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- GRGGYGGQGAGTTETTPAVSVKDGET PTDKVSVTIETV- KSWASRK-TVTLTVY- KALKVIGDKSWASRKGDTVTLTVYN TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- AKPTDKVSVTIETVGGAGQGGYGGLG DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- GQGAGQGAGTTETTPAVSVKDGETKA PTDKVSVTIETV- KVSVTIE-TET-TKA- LKVIGDKSWASRKGDTVTLTVYNAK TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- PTDKVSVTIETVGGAGQGGYGGLGSQ DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- GAGRGATTETTPAVSVKDGETKALKV PTDKVSVTIETV- KSWASRK-TVTLTVY- IGDKSWASRKGDTVTLTVYNAKPTD TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- KVSVTIETVGGAGQGGYGGLGGQGAG DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- RGAGTTETTPAVSVKDGETKALKVIG PTDKVSVTIETV- KVSVTIE DKSWASRKGDTVTLTVYNAKPTDKV TTETTPAVSVKDGETKALKVIG SVTIETVGGAGQGGYGGLGGQGAGRG DKSWASRKGDTVTLTVYNAK AGTTETTPAVSVKDGETKALKVIGDKS PTDKVSVTIETV- VVASRKGDTVTLTVYNAKPTDKVSVT TTETTPAVSVKDGETKALKVIG IETVQGGYGDLGSQGAGTTETTPAVSV DKSWASRKGDTVTLTVYNAK KDGETKALKVIGDKSVVASRKGDTVT PTDKVSVTIETV- LTVYNAKPTDKVSVTIETVGGAGQGG TTETTPAVSVKDGETKALKVIG YGGLGGQGAGRGAGTTETTPAVSVKD DKSWASRKGDTVTLTVYNAK GETKALKVIGDKSWASRKGDTVTLT PTDKVSVTIETV- VYNAKPTDKVSVTIETV TTETTPAVSVKDGETKALKVIG DKSWASRKGDTVTLTVYNAK PTDKVSVTIETV TTETTPAVSVKDGETKALKVIGDKSW TTETTPAVSVKDGETKALKVIG TET-TKALKVI-KSWASRK- ASRKGDTVTLTVYNAKPTDKVSVTIET DKSWASRKGDTVTLTVYNAK TVTLTVY-KVSVTIE-TET- VGGAGQGGYGGLGGQGAGQGAGTTE PTDKVSVTIETV- TKALKVI-KSWASRK- TTPAVSVKDGETKALKVIGDKSWAS TTETTPAVSVKDGETKALKVIG TVTLTVY-KVSVTIE-TET- RKGDTVTLTVYNAKPTDKVSVTIETV DKSWASRKGDTVTLTVYNAK TKALKVI-KSWASRK- GGAAQGGQGLGGQGTTETTPAVSVKD PTDKVSVTIETV- TVTLTVY-KVSVTIE-TET-TKA- GETKALKVIGDKSWASRKGDTVTLT TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- Dihedral - VYNAKPTDKVSVTIETVGGAGQGGYG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKALKVI- 47 D4 pA56735 GLGGQGAGQGAGTTETTPAVSVKDGE PTDKVSVTIETV- KSWASRK-TVTLTVY-

[0301]

[0302] TKALKVIGDKSWASRKGDTVTLTVY TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKA- 41

[0303] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0304] NAKPTDKVSVTIETVGGAGQGGFGGL DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- GGQGAGTTETTPAVSVKDGETKALKV PTDKVSVTIETV- KVSVTIE-TET-TKA- IGDKSWASRKGDTVTLTVYNAKPTD TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- KVSVTIETVGGAGQGGFGGLGGQGAG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- TTETTPAVSVKDGETKALKVIGDKSW PTDKVSVTIETV- KSWASRK-TVTLTVY- ASRKGDTVTLTVYNAKPTDKVSVTIET TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- VGGAAQGGQGLGGQGTTETTPAVSVK DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- DGETKALKVIGDKSWASRKGDTVTL PTDKVSVTIETV- KVSVTIE-TET-TKA- TVYNAKPTDKVSVTIETVGGAGQGGY TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- GGLGGQGAGRGAGTTETTPAVSVKDG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- ETKALKVIGDKSWASRKGDTVTLTV PTDKVSVTIETV- KSWASRK-TVTLTVY- YNAKPTDKVSVTIETVGQGGYGGLGG TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- QGAGRGAGTTETTPAVSVKDGETKAL DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- KVIGDKSWASRKGDTVTLTVYNAKP PTDKVSVTIETV- KVSVTIE TDKVSVTIETVGGAGQGGFGGLGGQG TTETTPAVSVKDGETKALKVIG AGTTETTPAVSVKDGETKALKVIGDKS DKSWASRKGDTVTLTVYNAK VVASRKGDTVTLTVYNAKPTDKVSVT PTDKVSVTIETV- IETVGGAGQGGYGGLGSQGAGRGGY TTETTPAVSVKDGETKALKVIG GGQGATTETTPAVSVKDGETKALKVI DKSWASRKGDTVTLTVYNAK GDKSWASRKGDTVTLTVYNAKPTDK PTDKVSVTIETV- VSVTIETVGGAGQGGYGGLGGQGAGR TTETTPAVSVKDGETKALKVIG GAGTTETTPAVSVKDGETKALKVIGD DKSWASRKGDTVTLTVYNAK KSWASRKGDTVTLTVYNAKPTDKVS PTDKVSVTIETV- VTIETV TTETTPAVSVKDGETKALKVIG DKSWASRKGDTVTLTVYNAK PTDKVSVTIETV TTETTPAVSVKDGETKALKVIGDKSW TTETTPAVSVKDGETKALKVIG TET-TKALKVI-KSWASRK- ASRKGDTVTLTVYNAKPTDKVSVTIET DKSWASRKGDTVTLTVYNAK TVTLTVY-KVSVTIE-TET- VGGAAQGGQGLGGQGTTETTPAVSVK PTDKVSVTIETV- TKALKVI-KSWASRK- DGETKALKVIGDKSWASRKGDTVTL TTETTPAVSVKDGETKALKVIG TVTLTVY-KVSVTIE-TET- TVYNAKPTDKVSVTIETVGGAGQGGF DKSWASRKGDTVTLTVYNAK TKALKVI-KSWASRK- GGLGGQGAGTTETTPAVSVKDGETKA PTDKVSVTIETV- TVTLTVY-KVSVTIE-TET-TKA- LKVIGDKSWASRKGDTVTLTVYNAK TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- PTDKVSVTIETVQGGYGDLGSQGAGT DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKALKVI- TETTPAVSVKDGETKALKVIGDKSW PTDKVSVTIETV- KSWASRK-TVTLTVY- Dihedral - ASRKGDTVTLTVYNAKPTDKVSVTIET TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKA- 48 D4 pA56736 VGGAGQGGYGGLGSQGAGRGGYGGQ DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY-

[0305]

[0306] GAGTTETTPAVSVKDGETKALKVIGD PTDKVSVTIETV- KVSVTIE-TET-TKA- 42

[0307] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0308] KSWASRKGDTVTLTVYNAKPTDKVS TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- VTIETVGGAAQGGQGLGGQGTTETTP DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- AVSVKDGETKALKVIGDKSWASRKG PTDKVSVTIETV- KSWASRK-TVTLTVY- DTVTLTVYNAKPTDKVSVTIETVGGA TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- GQGGYGGLGSQGAGRGGYGGQGAGT DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- TETTPAVSVKDGETKALKVIGDKSW PTDKVSVTIETV- KVSVTIE-TET-TKA- ASRKGDTVTLTVYNAKPTDKVSVTIET TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- VGGAGQGGYGGLGSQGAGRGGYGGQ DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- GAGTTETTPAVSVKDGETKALKVIGD PTDKVSVTIETV- KSWASRK-TVTLTVY- KSWASRKGDTVTLTVYNAKPTDKVS TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- VTIETVGGAGQGGFGGLGGQGAGTTE DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- TTPAVSVKDGETKALKVIGDKSWAS PTDKVSVTIETV- KVSVTIE RKGDTVTLTVYNAKPTDKVSVTIETV TTETTPAVSVKDGETKALKVIG GGAGQGGYGGLGGQGAGTTETTPAVS DKSWASRKGDTVTLTVYNAK VKDGETKALKVIGDKSWASRKGDTV PTDKVSVTIETV- TLTVYNAKPTDKVSVTIETVGGAGQG TTETTPAVSVKDGETKALKVIG GYGGLGGQGAGQGAGTTETTPAVSVK DKSWASRKGDTVTLTVYNAK DGETKALKVIGDKSWASRKGDTVTL PTDKVSVTIETV- TVYNAKPTDKVSVTIETVQGGYGDLG TTETTPAVSVKDGETKALKVIG SQGAGTTETTPAVSVKDGETKALKVIG DKSWASRKGDTVTLTVYNAK DKSWASRKGDTVTLTVYNAKPTDKV PTDKVSVTIETV- SVTIETV TTETTPAVSVKDGETKALKVIG DKSWASRKGDTVTLTVYNAK PTDKVSVTIETV TTETTPAVSVKDGETKALKVIGDKSW TTETTPAVSVKDGETKALKVIG TET-TKALKVI-KSWASRK- ASRKGDTVTLTVYNAKPTDKVSVTIET DKSWASRKGDTVTLTVYNAK TVTLTVY-KVSVTIE-TET- VGGAGQGGFGGLGGQGAGTTETTPAV PTDKVSVTIETV- TKALKVI-KSWASRK- SVKDGETKALKVIGDKSWASRKGDT TTETTPAVSVKDGETKALKVIG TVTLTVY-KVSVTIE-TET- VTLTVYNAKPTDKVSVTIETVGGAGQ DKSWASRKGDTVTLTVYNAK TKALKVI-KSWASRK- GGYGGLGGQGAGQGAGTTETTPAVSV PTDKVSVTIETV- TVTLTVY-KVSVTIE-TET-TKA- KDGETKALKVIGDKSVVASRKGDTVT TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- LTVYNAKPTDKVSVTIETVGGAGQGG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKALKVI- YGGLGSQGAGRGATTETTPAVSVKDG PTDKVSVTIETV- KSWASRK-TVTLTVY- ETKALKVIGDKSWASRKGDTVTLTV TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKA- YNAKPTDKVSVTIETVGGAGQGGYGG DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- Dihedral - LGGQGAGRGAGTTETTPAVSVKDGET PTDKVSVTIETV- KVSVTIE-TET-TKA- 49 D4 pA56737 KALKVIGDKSWASRKGDTVTLTVYN TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY-

[0309]

[0310] AKPTDKVSVTIETVGGAGQGGYGGLG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- 43

[0311] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0312] GQGAGQGAGTTETTPAVSVKDGETKA PTDKVSVTIETV- KSWASRK-TVTLTVY- LKVIGDKSWASRKGDTVTLTVYNAK TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- PTDKVSVTIETVGGAGQGGYGGLGGQ DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- GAGQGAGTTETTPAVSVKDGETKALK PTDKVSVTIETV- KVSVTIE-TET-TKA- VIGDKSWASRKGDTVTLTVYNAKPT TTETTPAVSVKDGETKALKVIG KSWASRK-TVTLTVY- DKVSVTIETVGQGGYGGLGGQGAGRG DKSWASRKGDTVTLTVYNAK KVSVTIE-TET-TKA- AGTTETTPAVSVKDGETKALKVIGDKS PTDKVSVTIETV- KSWASRK-TVTLTVY- VVASRKGDTVTLTVYNAKPTDKVSVT TTETTPAVSVKDGETKALKVIG KVSVTIE-TET-TKALKVI- IETVGQGGYGGLGGQGAGRGAGTTET DKSWASRKGDTVTLTVYNAK KSWASRK-TVTLTVY- TPAVSVKDGETKALKVIGDKSWASR PTDKVSVTIETV- KVSVTIE KGDTVTLTVYNAKPTDKVSVTIETVG TTETTPAVSVKDGETKALKVIG GAAQGGQGLGGQGTTETTPAVSVKDG DKSWASRKGDTVTLTVYNAK ETKALKVIGDKSWASRKGDTVTLTV PTDKVSVTIETV- YNAKPTDKVSVTIETVGGAGQGGYGG TTETTPAVSVKDGETKALKVIG LGSQGAGRGGYGGQGAGTTETTPAVS DKSWASRKGDTVTLTVYNAK VKDGETKALKVIGDKSWASRKGDTV PTDKVSVTIETV- TLTVYNAKPTDKVSVTIETVGGAAQG TTETTPAVSVKDGETKALKVIG GQGLGGQGTTETTPAVSVKDGETKAL DKSWASRKGDTVTLTVYNAK KVIGDKSWASRKGDTVTLTVYNAKP PTDKVSVTIETV- TDKVSVTIETV TTETTPAVSVKDGETKALKVIG DKSWASRKGDTVTLTVYNAK PTDKVSVTIETV LKYTIEVTES-KAEYVWK- ALKYTIE VTESGGKAEYVWKDNTGAI ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- LKAQKISNGDEVTVTTASGLKITVKAS NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- AQGGYGDLGSQGAGALKYTIEVTESG SGLKITVKASA- GKAEYVWKDNTGAILKAQKISNGDE ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- LKYTIEVTES-KAEYVWK- VTVTTASGLKITVKASAGGAGQGGYG NTGAILKAQKISNGDEVTVTTA GLGGQGAGQGAGALKYTIEVTESGGK SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- LKYTIEVTES-KAEYVWK- AEYVWKDNTGAILKAQKISNGDEVT ALKYTIE VTESGGKAEYVWKD VTTASGLKITVKASAGGAGQGGYGGL NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- LKYTIEVTES-KAEYVWK- GSQGAGRGGYGGQGAGALKYTIEVTE SGLKITVKASA- ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- SGGKAEYVWKDNTGAILKAQKISNG LKYTIEVTES-KAEYVWK- DEVTVTTASGLKITVKASAQGGYGDL NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- GSQGAGALKYTIEVTESGGKAEYVW SGLKITVKASA- LKYTIEVTES-KAEYVWK- KDNTGAILKAQKISNGDEVTVTTASGL ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- Dihedral - KITVKASAGGAGQGGFGGLGGQGAG NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- 50 D4 pA56738 ALKYTIE VTESGGKAEYVWKDNTGAI SGLKITVKASA- ILKAQK-EVTVT-KITVKAS

[0313]

[0314] LKAQKISNGDEVTVTTASGLKITVKAS ALKYTIE VTESGGKAEYVWKD

[0315] 44

[0316] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0317] AQGGYGDLGSQGAGALKYTIEVTESG NTGAILKAQKISNGDEVTVTTA GKAEYVWKDNTGAILKAQKISNGDE SGLKITVKASA- VTVTTASGLKITVKASAGGAGQGGYG ALKYTIEVTESGGKAEYVWKD GLGGQGAGQGAGALKYTIEVTESGGK NTGAILKAQKISNGDEVTVTTA AEYVWKDNTGAILKAQKISNGDEVT SGLKITVKASA- VTTASGLKITVKASA ALKYTIEVTESGGKAEYVWKD NTGAILKAQKISNGDEVTVTTA SGLKITVKASA ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- ALKYTIE VTESGGKAEYVWKDNTGAI NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- LKAQKISNGDEVTVTTASGLKITVKAS SGLKITVKASA- LKYTIEVTES-KAEYVWK- AGQGGYGGLGGQGAGRGAGALKYTI ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- EVTESGGKAEYVWKDNTGAILKAQK NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- ISNGDEVTVTTASGLKITVKASAGGAG SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- QGGYGGLGSQGAGRGGYGGQGAALK ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- YTIEVTESGGKAEYVWKDNTGAILKA NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- QKISNGDEVTVTTASGLKITVKASAGG SGLKITVKASA- LKYTIEVTES-KAEYVWK- AGQGGYGGLGGQGAGALKYTIEVTES ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- GGKAEYVWKDNTGAILKAQKISNGD NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- EVTVTTASGLKITVKASAGGAGQGGY SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- GGLGSQGAGRGGYGGQGAALKYTIEV ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- TESGGKAEYVWKDNTGAILKAQKIS NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- NGDEVTVTTASGLKITVKASAGGAGQ SGLKITVKASA- LKYTIEVTES-KAEYVWK- GGYGGLGSQGAGRGGYGGQGAALKY ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS TIEVTESGGKAEYVWKDNTGAILKAQ NTGAILKAQKISNGDEVTVTTA KISNGDEVTVTTASGLKITVKASAGGA SGLKITVKASA- AQGGQGLGGQGALKYTIEVTESGGKA ALKYTIE VTESGGKAEYVWKD EYVWKDNTGAILKAQKISNGDEVTV NTGAILKAQKISNGDEVTVTTA TTASGLKITVKASAGGAGQGGYGGLG SGLKITVKASA- SQGAGRGAALKYTIEVTESGGKAEYV ALKYTIE VTESGGKAEYVWKD

[0318] Dihedral - VVKDNTGAILKAQKISNGDEVTVTTAS NTGAILKAQKISNGDEVTVTTA

[0319] 51 D4 pA56739 GLKITVKASA SGLKITVKASA

[0320] ALKYTIE VTESGGKAEYVWKDNTGAI ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- LKAQKISNGDEVTVTTASGLKITVKAS NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- AGGAGQGGFGGLGGQGAGALKYTIEV SGLKITVKASA- LKYTIEVTES-KAEYVWK- Dihedral - TESGGKAEYVWKDNTGAILKAQKIS ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- 52 D4 pA56741

[0321]

[0322] NGDEVTVTTASGLKITVKASAGGAAQ NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- 45

[0323] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0324] GGQGLGGQGALKYTIEVTESGGKAEy SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- VWKDNTGAILKAQKISNGDEVTVTT ALKYTIEVTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- ASGLKITVKASAGGAGQGGYGGLGSQ NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- GAGRGGYGGQGAGALKYTIEVTESGG SGLKITVKASA- LKYTIEVTES-KAEYVWK- KAEYVWKDNTGAILKAQKISNGDEV ALKYTIEVTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- TVTTASGLKITVKASAGQGGYGGLGG NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- QGAGRGAGALKYTIEVTESGGKAEYV SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- VVKDNTGAILKAQKISNGDEVTVTTAS ALKYTIEVTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- GLKITVKASAGGAGQGGYGGLGGQG NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- AGALKYTIEVTESGGKAEYVWKDNT SGLKITVKASA- LKYTIEVTES-KAEYVWK- GAILKAQKISNGDEVTVTTASGLKITV ALKYTIEVTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS KASAGGAGQGGYGGLGSQGAGRGGY NTGAILKAQKISNGDEVTVTTA GGQGAALKYTIEVTESGGKAEYVWK SGLKITVKASA- DNTGAILKAQKISNGDEVTVTTASGLK ALKYTIEVTESGGKAEYVWKD ITVKASAGQGGYGGLGGQGAGRGAG NTGAILKAQKISNGDEVTVTTA ALKYTIE VTESGGKAEYVWKDNTGAI SGLKITVKASA- LKAQKISNGDEVTVTTASGLKITVKAS ALKYTIEVTESGGKAEYVWKD A NTGAILKAQKISNGDEVTVTTA SGLKITVKASA ALKYTIE VTESGGKAEYVWKDNTGAI ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- LKAQKISNGDEVTVTTASGLKITVKAS NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- AGGAGQGGYGGLGSQGAGRGAALKY SGLKITVKASA- LKYTIEVTES-KAEYVWK- TIEVTESGGKAEYVWKDNTGAILKAQ ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- KISNGDEVTVTTASGLKITVKASAGGA NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- GQGGYGGLGSQGAGRGGYGGQGAAL SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- KYTIEVTESGGKAEYVWKDNTGAILK ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- AQKISNGDEVTVTTASGLKITVKASAG NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- GAGQGGYGGLGGQGAGALKYTIEVTE SGLKITVKASA- LKYTIEVTES-KAEYVWK- SGGKAEYVWKDNTGAILKAQKISNG ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS- DEVTVTTASGLKITVKASAGGAGQGG NTGAILKAQKISNGDEVTVTTA LKYTIEVTES-KAEYVWK- FGGLGGQGAGALKYTIEVTESGGKAE SGLKITVKASA- ILKAQK-EVTVT-KITVKAS- YVWKDNTGAILKAQKISNGDEVTVT ALKYTIE VTESGGKAEYVWKD LKYTIEVTES-KAEYVWK- TASGLKITVKASAGQGGYGGLGGQGA NTGAILKAQKISNGDEVTVTTA ILKAQK-EVTVT-KITVKAS- GRGAGALKYTIEVTESGGKAEYVWK SGLKITVKASA- LKYTIEVTES-KAEYVWK- DNTGAILKAQKISNGDEVTVTTASGLK ALKYTIE VTESGGKAEYVWKD ILKAQK-EVTVT-KITVKAS ITVKASAGQGGYGGLGGQGAGRGAG NTGAILKAQKISNGDEVTVTTA

[0325] Dihedral - ALKYTIE VTESGGKAEYVWKDNTGAI SGLKITVKASA- 53 D4 pA56742

[0326]

[0327] LKAQKISNGDEVTVTTASGLKITVKAS ALKYTIE VTESGGKAEYVWKD

[0328] 46

[0329] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0330] AGGAGQGGYGGLGGQGAGQGAGAL NTGAILKAQKISNGDEVTVTTA KYTIEVTESGGKAEYVWKDNTGAILK SGLKITVKASA- AQKISNGDEVTVTTASGLKITVKASA ALKYTIEVTESGGKAEYVWKD NTGAILKAQKISNGDEVTVTTA SGLKITVKASA MFKVKAIGNTRAVIIGEGLLEL VEPGE FKVKA-TRAVIIG-LLELV- ELEVEATGLVTVLVIPLPSGGSGPQGP ELEV-VTVLVIP-FKVKA- GSQGPYGPGSQGPGSQGPYGPGSQGP TRAVIIG-LLELV-ELEV- GSQGPGSQGPGSQGPYGPGSQGPGGA VTVLVIP-FKVKA-TRAVIIG- SASMFKVKAIGNTRAVIIGEGLLELVEP LLELV-ELEV-VTVLVIP- GEELEVEATGLVTVLVIPLPSGGSGPQ FKVKA-TRAVIIG-LLELV- GPGSQGPYGPGSQGPGSQGPYGPGSQ ELEV-VTVLVIP-FKVKA- GPGSQGPGSQGPGSQGPYGPGSQGPG TRAVIIG-LLELV-ELEV- GASASMFKVKAIGNTRAVIIGEGLLEL MFKVKAIGNTRAVIIGEGLLEL VTVLVIP-FKVKA-TRAVIIG- VEPGEELEVEATGLVTVLVIPLPSGGSG VEPGEELEVEATGLVTVLVIPLP LLELV-ELEV-VTVLVIP- PQGPGSQGPYGPGSQGPGSQGPYGPGS FKVKA-TRAVIIG-LLELV- QGPGSQGPGSQGPGSQGPYGPGSQGP MFKVKAIGNTRAVIIGEGLLEL ELEV-VTVLVIP-FKVKA- GGASASMFKVKAIGNTRAVIIGEGLLE VEPGEELEVEATGLVTVLVIPLP TRAVIIG-LLELV-ELEV- LVEPGEELEVEATGLVTVLVIPLPSGGS VTVLVIP GPQGPGSQGPYGPGSQGPGSQGPYGP MFKVKAIGNTRAVIIGEGLLEL GSQGPGSQGPGSQGPGSQGPYGPGSQ VEPGEELEVEATGLVTVLVIPLP GPGGASASMFKVKAIGNTRAVIIGEGL LELVEPGEELEVEATGLVTVLVIPLPSG MFKVKAIGNTRAVIIGEGLLEL GSGPQGPGSQGPYGPGSQGPGSQGPY VEPGEELEVEATGLVTVLVIPLP GPGSQGPGSQGPGSQGPGSQGPYGPGS QGPGGASASMFKVKAIGNTRAVIIGEG MFKVKAIGNTRAVIIGEGLLEL LLELVEPGEELEVEATGLVTVLVIPLPS VEPGEELEVEATGLVTVLVIPLP GGSGPQGPGSQGPYGPGSQGPGSQGP YGPGSQGPGSQGPGSQGPGSQGPYGP MFKVKAIGNTRAVIIGEGLLEL GSQGPGGASASMFKVKAIGNTRAVIIG VEPGEELEVEATGLVTVLVIPLP EGLLELVEPGEELEVEATGLVTVLVIPL PSGGSGPQGPGSQGPYGPGSQGPGSQG MFKVKAIGNTRAVIIGEGLLEL PYGPGSQGPGSQGPGSQGPGSQGPYGP VEPGEELEVEATGLVTVLVIPLP GSQGPGGASASMFKVKAIGNTRAVIIG

[0331] Dihedral - EGLLELVEPGEELEVEATGLVTVLVIPL MFKVKAIGNTRAVIIGEGLLEL

[0332] 54 D4 pA56756 P VEPGEELEVEATGLVTVLVIPLP

[0333]

[0334] 47

[0335] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0336] AKLYTVEVTSSGGAATHVWK KLYTVEVTSS-AATHVWK- AKLYTVEVTSSGGAATHVWKDEVGA DEVGAILKAQAVSNGDTVTVTT ILKAQA-TVTVT-KITVTAS- ILKAQAVSNGDTVTVTTADGLKITVTA ADGLKITVTASA- KLYTVEVTSS-AATHVWK- SAGQGGYGGLGGQGAGRGAGAKLYT AKLYTVEVTSSGGAATHVWK ILKAQA-TVTVT-KITVTAS- VEVTSSGGAATHVWKDEVGAILKAQ DEVGAILKAQAVSNGDTVTVTT KLYTVEVTSS-AATHVWK- AVSNGDTVTVTTADGLKITVTASAGG ADGLKITVTASA- ILKAQA-TVTVT-KITVTAS- AGQGGYGGLGSQGAGRGGYGGQGAA AKLYTVEVTSSGGAATHVWK KLYTVEVTSS-AATHVWK- KLYTVEVTSSGGAATHVWKDEVGAI DEVGAILKAQAVSNGDTVTVTT ILKAQA-TVTVT-KITVTAS- LKAQAVSNGDTVTVTTADGLKITVTA ADGLKITVTASA- KLYTVEVTSS-AATHVWK- SAGGAGQGGYGGLGGQGAGAKLYTV AKLYTVEVTSSGGAATHVWK ILKAQA-TVTVT-KITVTAS- EVTSSGGAATHVWKDEVGAILKAQA DEVGAILKAQAVSNGDTVTVTT KLYTVEVTSS-AATHVWK- VSNGDTVTVTTADGLKITVTASAGGA ADGLKITVTASA- ILKAQA-TVTVT-KITVTAS- GQGGYGGLGSQGAGRGGYGGQGAAK AKLYTVEVTSSGGAATHVWK KLYTVEVTSS-AATHVWK- LYTVEVTSSGGAATHVWKDEVGAIL DEVGAILKAQAVSNGDTVTVTT ILKAQA-TVTVT-KITVTAS- KAQAVSNGDTVTVTTADGLKITVTAS ADGLKITVTASA- KLYTVEVTSS-AATHVWK- AGGAGQGGYGGLGSQGAGRGGYGGQ AKLYTVEVTSSGGAATHVWK ILKAQA-TVTVT-KITVTAS GAAKLYTVEVTSSGGAATHVWKDEV DEVGAILKAQAVSNGDTVTVTT GAILKAQAVSNGDTVTVTTADGLKITV ADGLKITVTASA- TASAGGAAQGGQGLGGQGAKLYTVE AKLYTVEVTSSGGAATHVWK VTSSGGAATHVWKDEVGAILKAQAV DEVGAILKAQAVSNGDTVTVTT SNGDTVTVTTADGLKITVTASAGGAG ADGLKITVTASA- QGGYGGLGSQGAGRGAAKLYTVEVT AKLYTVEVTSSGGAATHVWK

[0337] Dihedral - SSGGAATHVWKDEVGAILKAQAVSN DEVGAILKAQAVSNGDTVTVTT

[0338] 80 D4 pA60317 GDTVTVTTADGLKITVTASA ADGLKITVTASA

[0339] AKLYTVEVTESGGKATHVWRDEVGA AKLYTVEVTESGGKATHVWR KLYTVEVTES-KATHVWR- ILKAQAVSNGDTVTVTTADGLKISVKA DEVGAILKAQAVSNGDTVTVTT ILKAQA-TVTVT-KISVKAS- SAGQGGYGGLGGQGAGRGAGAKLYT ADGLKISVKASA- KLYTVEVTES-KATHVWR- VEVTESGGKATHVWRDEVGAILKAQ AKLYTVEVTESGGKATHVWR ILKAQA-TVTVT-KISVKAS- AVSNGDTVTVTTADGLKISVKASAGG DEVGAILKAQAVSNGDTVTVTT KLYTVEVTE-ATHVWR- AGQGGYGGLGSQGAGRGGYGGQGAA ADGLKISVKASA- ILKAQA-TVTVT-KISVKAS- KLYTVEVTESGGKATHVWRDEVGAI AKLYTVEVTESGGKATHVWR KLYTVEVTE-ATHVWR- LKAQAVSNGDTVTVTTADGLKISVKA DEVGAILKAQAVSNGDTVTVTT ILKAQA-TVTVT-KISVKAS- SAGGAGQGGYGGLGGQGAGAKLYTV ADGLKISVKASA- KLYTVEVTE-ATHVWR- EVTESGGKATHVWRDEVGAILKAQA AKLYTVEVTESGGKATHVWR ILKAQA-TVTVT-KISVKAS- VSNGDTVTVTTADGLKISVKASAGGA DEVGAILKAQAVSNGDTVTVTT KLYTVEVTES-KATHVWR- Dihedral - GQGGYGGLGSQGAGRGGYGGQGAAK ADGLKISVKASA- ILKAQA-TVTVT-KISVKAS- 81 D4 pA60319 LYTVEVTESGGKATHVWRDEVGAIL AKLYTVEVTESGGKATHVWR KLYTVEVTE-ATHVWR-

[0340]

[0341] KAQAVSNGDTVTVTTADGLKISVKAS DEVGAILKAQAVSNGDTVTVTT ILKAQA-TVTVT-KISVKAS- 48

[0342] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0343] AGGAGQGGYGGLGSQGAGRGGYGGQ ADGLKISVKASA- KLYTVEVTE-ATHVWR- GAAKLYTVEVTESGGKATHVWRDEV AKLYTVEVTESGGKATHVWR ILKAQA-TVTVT-KISVKAS GAILKAQAVSNGDTVTVTTADGLKISV DEVGAILKAQAVSNGDTVTVTT KASAGGAAQGGQGLGGQGAKLYTVE ADGLKISVKASA- VTESGGKATHVWRDEVGAILKAQAV AKLYTVEVTESGGKATHVWR SNGDTVTVTTADGLKISVKASAGGAG DEVGAILKAQAVSNGDTVTVTT QGGYGGLGSQGAGRGAAKLYTVEVT ADGLKISVKASA- ESGGKATHVWRDEVGAILKAQAVSN AKLYTVEVTESGGKATHVWR GDTVTVTTADGLKISVKASA DEVGAILKAQAVSNGDTVTVTT ADGLKISVKASA AKLYTVKVTTSNGKAEYWIED KLYTVKVTTS-KAEYWIE- AKLYTVKVTTSNGKAEYVVIEDEVGAI EVGAILKAQSVSNGDEVTVTTA ILKAQS-EVTVT-KITVKAS- LKAQSVSNGDEVTVTTADGLKITVKA DGLKITVKASA- KLYTVKVTTS-KAEYWIE- SAGQGGYGGLGGQGAGRGAGAKLYT AKLYTVKVTTSNGKAEYWIED ILKAQS-EVTVT-KITVKAS- VKVTTSNGKAEYWIEDEVGAILKAQS EVGAILKAQSVSNGDEVTVTTA KLYTVKVTTS-KAEYWIE- VSNGDEVTVTTADGLKITVKASAGGA DGLKITVKASA- ILKAQS-EVTVT-KITVKAS- GQGGYGGLGSQGAGRGGYGGQGAAK AKLYTVKVTTSNGKAEYWIED KLYTVKVTTS-KAEYWIE- LYTVKVTTSNGKAEYWIEDEVGAILK EVGAILKAQSVSNGDEVTVTTA ILKAQS-EVTVT-KITVKAS- AQSVSNGDEVTVTTADGLKITVKASA DGLKITVKASA- KLYTVKVTTS-KAEYWIE- GGAGQGGYGGLGGQGAGAKLYTVKV AKLYTVKVTTSNGKAEYWIED ILKAQS-EVTVT-KITVKAS- TTSNGKAEYWIEDEVGAILKAQSVSN EVGAILKAQSVSNGDEVTVTTA KLYTVKVTTS-KAEYWIE- GDEVTVTTADGLKITVKASAGGAGQG DGLKITVKASA- ILKAQS-EVTVT-KITVKAS- GYGGLGSQGAGRGGYGGQGAAKLYT AKLYTVKVTTSNGKAEYWIED KLYTVKVTTS-KAEYWIE- VKVTTSNGKAEYWIEDEVGAILKAQS EVGAILKAQSVSNGDEVTVTTA ILKAQS-EVTVT-KITVKAS- VSNGDEVTVTTADGLKITVKASAGGA DGLKITVKASA- KLYTVKVTTS-KAEYWIE- GQGGYGGLGSQGAGRGGYGGQGAAK AKLYTVKVTTSNGKAEYWIED ILKAQS-EVTVT-KITVKAS LYTVKVTTSNGKAEYWIEDEVGAILK EVGAILKAQSVSNGDEVTVTTA AQSVSNGDEVTVTTADGLKITVKASA DGLKITVKASA- GGAAQGGQGLGGQGAKLYTVKVTTS AKLYTVKVTTSNGKAEYWIED NGKAEYWIEDEVGAILKAQSVSNGD EVGAILKAQSVSNGDEVTVTTA EVTVTTADGLKITVKASAGGAGQGGY DGLKITVKASA- GGLGSQGAGRGAAKLYTVKVTTSNG AKLYTVKVTTSNGKAEYWIED

[0344] Dihedral - KAEYWIEDEVGAILKAQSVSNGDEVT EVGAILKAQSVSNGDEVTVTTA

[0345] 82 D4 pA60321 VTTADGLKITVKASA DGLKITVKASA

[0346] Dihedral - AKLYTVKVTTSNGKAEYVVIEDEVGAI AKLYTVKVTTSNGKAEYWIED KLYTVKVTTS-KAEYWIE- 83 D4 pA60320 LKAQSVSNGDEVTVTTADGLKITVKA EVGAILKAQSVSNGDEVTVTTA ILKAQS-EVTVT-KITVKAS-

[0347]

[0348] SAGGAGQGGYGGLGSQGAGRGGYGG DGLKITVKASA- KLYTVKVTTS-KAEYWIE- 49

[0349] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0350] QGAAKLYTVKVTTSNGKAEYWIEDE AKLYTVKVTTSNGKAEYWIED ILKAQS-EVTVT-KITVKAS- VGAILKAQSVSNGDEVTVTTADGLKIT EVGAILKAQSVSNGDEVTVTTA KLYTVKVTTS-KAEYWIE- VKASAGGAAQGGQGLGGQGAKLYTV DGLKITVKASA- ILKAQS-EVTVT-KITVKAS- KVTTSNGKAEYWIEDEVGAILKAQSV AKLYTVKVTTSNGKAEYWIED KLYTVKVTTS-KAEYWIE- SNGDEVTVTTADGLKITVKASAGGAG EVGAILKAQSVSNGDEVTVTTA ILKAQS-EVTVT-KITVKAS- QGGYGGLGGQGAGRGAGAKLYTVKV DGLKITVKASA- KLYTVKVTTS-KAEYWIE- TTSNGKAEYWIEDEVGAILKAQSVSN AKLYTVKVTTSNGKAEYWIED ILKAQS-EVTVT-KITVKAS- GDEVTVTTADGLKITVKASAGGAAQG EVGAILKAQSVSNGDEVTVTTA KLYTVKVTTS-KAEYWIE- GQGLGGQGAKLYTVKVTTSNGKAEY DGLKITVKASA- ILKAQS-EVTVT-KITVKAS- VVIEDEVGAILKAQSVSNGDEVTVTTA AKLYTVKVTTSNGKAEYWIED KLYTVKVTTS-KAEYWIE- DGLKITVKASAGQGGYGGLGGQGAG EVGAILKAQSVSNGDEVTVTTA ILKAQS-EVTVT-KITVKAS- RGAGAKLYTVKVTTSNGKAEYVVIED DGLKITVKASA- KLYTVKVTTS-KAEYWIE- EVGAILKAQSVSNGDEVTVTTADGLKI AKLYTVKVTTSNGKAEYWIED ILKAQS-EVTVT-KITVKAS TVKASAGGAGQGGFGGLGGQGAGAK EVGAILKAQSVSNGDEVTVTTA LYTVKVTTSNGKAEYWIEDEVGAILK DGLKITVKASA- AQSVSNGDEVTVTTADGLKITVKASA AKLYTVKVTTSNGKAEYWIED GGAGQGGYGGLGGQGAGQGAGAKL EVGAILKAQSVSNGDEVTVTTA YTVKVTTSNGKAEYWIEDEVGAILKA DGLKITVKASA- QSVSNGDEVTVTTADGLKITVKASA AKLYTVKVTTSNGKAEYWIED EVGAILKAQSVSNGDEVTVTTA DGLKITVKASA

[0351]

[0352] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0353] IL Beta strand reservoir polypeptides Exhibiting a Tertiary Beta Solenoid or Antiparallel Structure

[0354]

[0168] The disclosure provides non-naturally occurring beta strand reservoir polypeptides that are determined based on in silico analyses to exhibit a tertiary beta solenoid structure. In embodiments, the beta strand reservoir polypeptides disclosed herein comprise at least two beta strands per beta sheet (for instance, 3 beta strands, 4 beta strands, 5 beta strands, 6 beta strands, 7 beta strands, 8 beta strands, 9 beta strands, 10 beta strands, 11 beta strands, 12 beta strands, 13 beta strands, 14 beta strands, 15 beta strands, 16 beta strands, 17 beta strands, 18 beta strands, 19 beta strands, 20 beta strands or more) and a linker between each of the beta strands.

[0355]

[0169] In embodiments, the beta strands associate to form two parallel beta sheets (B 1 and B2) such that the beta strand reservoir polypeptide exhibits a beta solenoid structure. Exemplary beta strand reservoir polypeptides exhibiting a beta solenoid structure based on in silico analyses are shown in FIGs. 5A, 5B, 6A, 6B, 7A and 7B. In embodiments, one or more Ca carbons of beta sheet Bl are separated by a distance in the range of about 6A to about 12A from one or more Ca carbons of beta sheet B2, for example, about 6 A, about 7 A, about 8 A, about 9 A, about 10 A, about 11 A, or about 12A. In embodiments, one or more Ca carbons of beta sheet Bl are separated by about 12A from one or more Ca carbons of beta sheet B2. In embodiments, the beta strands in beta sheet Bl are pointed in a direction opposite to the beta strands of beta sheet B2. In embodiments, the beta strand reservoir polypeptide comprising a beta solenoid structure comprises: (L - S - L -S)x- L, wherein L is a linker, S is a beta strand, x > 2, and the L - S - L - S comprises about 18 to about 92 amino acids in length.

[0356]

[0170] In embodiments, one or both of beta sheet Bl and beta sheet B2 comprises at least two and up to 20 beta strands. In embodiments, one or more of beta sheet Bl and beta sheet B2 comprises more than 20 beta strands. In embodiments, each of beta sheet Bl and beta sheet B2 comprises at least two and up to 20 beta strands, for example, about 3 beta strands, about 4 beta strands, about 5 beta strands, about 6 beta strands, about 7 beta strands, about 8 beta strands, about 9 beta strands, about 10 beta strands, about 11 beta strands, about 12 beta strands, about 13 beta strands, about 14 beta strands, about 15 beta strands, about 16 beta strands, about 17 beta strands, about 18 beta strands, about 19 beta strands, or about 20 beta strands. In embodiments, each of beta sheet Bl and

[0357] 51

[0358] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0359] beta sheet B2 comprises at least 5 and up to 16 beta strands. In embodiments, the polypeptide comprises at least 100 amino acid residues, for instance, about 110 amino acid residues, about 120 amino acid residues, about 130 amino acid residues, about 140 amino acid residues, about 150 amino acid residues, about 160 amino acid residues, about 170 amino acid residues, about 180 amino acid residues, about 190 amino acid residues, about 200 amino acid residues, or more than 200 amino acid residues. In embodiments, the polypeptide comprises at least 200 amino acid residues, for instance, about 250 amino acid residues, about 300 amino acid residues, about 350 amino acid residues, about 400 amino acid residues, about 450 amino acid residues, about 500 amino acid residues. In embodiments, the polypeptide comprises about 300 amino acids to about 500 amino acid residues.

[0360]

[0171] In embodiments, beta sheet B 1 and beta sheet B2 have the same number of beta strands. In embodiments, beta sheet Bl and beta sheet B2 have different numbers of beta strands.

[0361]

[0172] In embodiments, a beta sheet comprises beta strands that exhibit pairwise sequence identity of at least about 30%, for instance, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween and excluding beta strands that are present in capping domains. For instance, in embodiments, if a beta sheet comprises 16 beta strands (SI through SI 6), excluding beta strands that are present in capping domains, then the pairwise sequence identity among the 16 beta strands (e.g., SI v. S2, SI v. S3, SI v. S4, S2 v. S3 and so on) is at least about 30%, for instance, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween.

[0362]

[0173] In embodiments, 80% of beta strands belonging within a beta domain or beta solenoid motif do not comprise proline. In embodiments, the beta strands of beta sheet Bl and beta sheet B2 do not comprise proline.

[0363]

[0174] In embodiments, the beta strands of beta sheet Bl and beta sheet B2 are about 3 amino acid residues to about 26 amino acid residues in length, for instance, 3 amino acid residues, 4 amino 52

[0364] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0365] acid residues, 5 amino acid residues, 6 amino acid residues, 7 amino acid residues, 8 amino acid residues 9 amino acid residues, 10 amino acid residues, 11 amino acid residues, 12 amino acid residues, 13 amino acid residues, 14 amino acid residues, 15 amino acid residues, 16 amino acid residues, 17 amino acid residues, 18 amino acid residues, 19 amino acid residues, 20 amino acid residues, 21 amino acid residues, 22 amino acid residues, 23 amino acid residues, 24 amino acid residues, 25 amino acid residues or 26 amino acid residues. In embodiments, the beta strands of beta sheet B 1 and beta sheet B2 are about 5 amino acid residues to about 7 amino acid residues in length, for instance, 5 amino acid residues, 6 amino acid residues, or 7 amino acid residues.

[0366]

[0175] In embodiments, the beta strands of beta sheet Bl and / or beta sheet B2 exhibit an average pLDDT of over 70. In embodiments, the beta strands of beta sheet B 1 and / or beta sheet B2 exhibit an average pLDDT of over 90. In embodiments, the beta strands of beta sheet B 1 and / or beta sheet B2 exhibit an average pLDDT of over 70, for instance over 75, over 80, over 85, over 90, or over 95. In embodiments, the beta strands of beta sheet Bl and / or beta sheet B2 exhibit an average pLDDT of over 90. In embodiments, the beta strands of beta sheet B 1 and / or beta sheet B2 exhibit an average pLDDT that lies in the range of about 50 to about 100, for example about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, or about 95. In embodiments, the beta strands of beta sheet Bl and / or beta sheet B2 exhibit an average pLDDT that lies in the range of about 50 to about 100, for example about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, or about 95. In embodiments, the beta strands of beta sheet Bl and / or beta sheet B2 exhibit an average pLDDT that lies in the range of about 50 to about 70. In embodiments, the beta strands of beta sheet Bl and / or beta sheet B2 exhibit an average pLDDT that lies in the range of about 50 to about 70. In embodiments, the beta strands of beta sheet Bl and / or beta sheet B2 exhibit an average pLDDT that lies in the range of about 70 to about 100.

[0367]

[0176] In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein has any one of the types of structures depicted in FIGs. 8A-8L. In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein comprises two beta sheets connected via linkers (referred to herein as a type #1 beta solenoid). In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein comprises two beta sheets connected via loops (referred to herein as a type #2 beta solenoid). In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein is a type #3 beta solenoid comprising two type #2 solenoids connected via a linker.

[0368] 53

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[0370] In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein is a type #4 beta solenoid comprising two beta sheets connected via loops except when they are connected via a linker after odd numbered revolutions (e.g. 3 or 5 revolutions) of the beta solenoid. In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein is a type #5 beta solenoid in which the two beta sheets are connected via a linker comprising or consisting of an amino acid sequence of 8 amino acids or less, that is at least 60% glycine (such as SEQ ID NO: 147 or 148). In embodiments, a BSRP exhibiting a beta solenoid tertiary structure disclosed herein is a type #6 beta solenoid comprising two beta sheets connected, in an alternating manner, via (i) a linker comprising or consisting of an amino acid sequence of 8 amino acids or less, that is at least 60% glycine (such as SEQ ID NO: 147 or 148), and (ii) a linker not comprising or consisting of the amino acid sequence of SEQ ID NO: 147 or SEQ ID NO: 148, as depicted in FIGs. 8K-8L.

[0371]

[0177] In embodiments, one or more of the beta strands comprises an amino acid sequence that is at least 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to any one of SEQ ID NOs: 597-787 and 789-1053. In embodiments, each of the beta strands comprises an amino acid sequence that is at least 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to any one of SEQ ID NOs: 597-787 and 789-1053.

[0372]

[0178] In embodiments, the beta strand reservoir polypeptides having a beta solenoid structure comprise a domain structure illustrated in FIGs. 8A and 8B. In other words, the beta strand reservoir polypeptides having a beta solenoid structure comprise a plurality of beta solenoid motifs linked together by linkers. In embodiments, the beta solenoid motif comprises a consensus amino acid sequence of any one of SEQ ID NOs: 1590-1600, and 1602-1655. In embodiments, the beta solenoid motif comprises an amino acid sequence that is at least 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of 1146-1259, 1261, and 1264-1589.

[0373] 54

[0374] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0375]

[0179] In embodiments, the beta strand reservoir polypeptides that are determined to exhibit a tertiary beta solenoid structure comprise an amino acid sequence that is at least 50% (for instance, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of any one of SEQ ID NOs: 1, 4-14, 16-45, and 55-79.

[0376]

[0180] In embodiments, beta strand reservoir polypeptides that are determined to exhibit a tertiary beta solenoid structure comprise a capping domain at the N-terminus and / or the C-terminus. This reduces the risk of the N-terminus of one polypeptide interacting with the C-terminus of another polypeptide. As used herein, a “capping domain” is a stretch of amino acids present at the N-terminus or the C-terminus of a BSRP having a beta solenoid structure or an anti-parallel beta sheet structure disclosed herein. In embodiments, the capping domain is at least 4 or at least 5 amino acids, for instance, about 15 amino acids. In embodiments, the capping domain at the N-terminus spans from the N-terminus to the amino acid right before the first instance of the consensus beta solenoid motif found in the solenoid that the capping domain is part of. In embodiments, the capping domain at the C-terminus spans from the C-terminus to the amino acid right after the last amino acid of the most C-terminal instance of the consensus beta solenoid motif found in the solenoid that the capping domain is part of. In embodiments, the capping domain comprises a beta strand. In embodiments, the capping domain does not comprise the consensus beta solenoid motif found in the solenoid that the capping domain is part of (one of SEQ ID NOs: 1590-1600, 1602-1655). In embodiments, the length of the amino acid sequence of the capping domain is about 1% to about 30% of the length of the beta strand reservoir polypeptide. Without being bound by theory, it is thought that the presence of a capping domain at the N-terminus and / or the C-terminus prevents interactions between the N-terminus and C-terminus of different individual beta strand reservoir polypeptides, which could be detrimental to protein expression due to the possibility of forming unbounded assemblies within the host organism or within the growth media, in the case of protein secretion.

[0377]

[0181] In embodiments, the capping domain at the N-terminus comprises an amino acid sequence having at least about 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive 55

[0378] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0379] of all values and subranges that lie therebetween) identity to any one of SEQ ID NOs: 1656-1708. In embodiments, the capping domain at the N-terminus comprises or consists of an amino acid sequence of any one of SEQ ID NOs: 1656-1708.

[0380]

[0182] In embodiments, the capping domain at the C-terminus comprises an amino acid sequence having at least about 80% (for instance, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identity to any one of SEQ ID NOs: 1709-1774. In embodiments, the capping domain at the C-terminus comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 1709-1774.

[0381]

[0183] In embodiments, the beta strand reservoir polypeptides disclosed herein fold into antiparallel beta sheets. In embodiments, the beta strand reservoir polypeptide that is determined to exhibit an anti-parallel beta sheet structure comprises an amino acid sequence that is at least 50% (for instance, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) identical to the amino acid sequence of SEQ ID NO: 15. In embodiments, the beta strand reservoir polypeptide that is determined to exhibit an anti-parallel beta sheet structure comprises a beta strand comprising an amino acid sequence of any one of SEQ ID NOs: 739, 738, 2101, 2102, 735, 736, 846, 845, 844, 737, 1060, and 2153. Without being bound by theory, it is thought that the formation of the beta solenoid vs. anti-parallel beta sheet structure might be promoted by the different linker sequences.

[0382]

[0184] In embodiments, the beta strands associate to form one anti-parallel beta sheet. In embodiments, the beta strand reservoir polypeptide comprising an anti-parallel beta sheet structure comprises: (L - S - L - S)x- L, wherein L is a linker, S is a beta strand, x > 2, and the L - S - L - S comprises about 18 to about 92 amino acids in length.

[0383]

[0185] Table 2 lists the beta strand reservoir polypeptides that are determined based on in silico analyses to exhibit a tertiary beta solenoid structure disclosed herein along with their respective beta solenoid motifs and beta strands within beta solenoid motifs and capping domains. The SEQ ID NO. in the first column applies to the full sequence listed in the fourth column of the table. In embodiments, the present disclosure provides a sequence according to any of SEQ ID NO: 4, 16,

[0384] 56

[0385] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0386] 19, 25, 61, 62, 64, 65, 76, 77, or 78, followed by a W (Trp). In embodiments, the present disclosure provides a sequence according to any of SEQ ID NO: 5, 35, or 37, followed by a GSGS (Gly-Ser-Gly-Ser). In embodiments, the present disclosure provides an amino acid sequence of Table 2 having an initial methionine. In embodiments, the present disclosure provides an amino acid sequence of Table 2 that does not comprise an initial methionine.

[0387] 57

[0388] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0389] Table 2

[0390] SEQ Structure Name Full Sequence Beta solenoid motifs (separated by a Beta strands (separated by a ID of beta dash dash

[0391] NO: domains

[0392] in silico

[0393] 1 Parallel pA55724 MNKIKKTKNSEVKSTGENNEIESKSKKG ENKIESTGDNNKIEST- KIKK-EVKSTG-EIESK- solenoid KNKIESTGGAGQGGYGGLGGQGAGRG KNKIESTGDKNKIEST- KIESTG-ENKIEST-KIEST- AGGNENKIESTGDNNKIESTGGAGQGG ENKIESKGNENKIESTG- KNKffiST-KIEST-ENKIESK- FGGLGGQGAGGNKNKIESTGDKNKIES KNKIESKGDKNKIESTQ- KIESTG-KNKIESK-KIESTQ- TGGAGQGGFGGLGGQGAGGNENKIESK ENKIESKGDKNKIESK- ENKIESK-KIESK-KNKIESK- GNENKIESTGGAGQGGYGGLGSQGAGR KNKIESKGNKNKIESK- KIESK-KNKIESK-KIESK- GGYGGQGAGNKNKIESKGDKNKIESTQ KNKIESKGNKNKIESK- ENKIESK-KIESK- GGYGDLGSQGAGGNENKIESKGDKNKI ENKIESKGNENKIESK KNKIESTGNKNKIESE- ESKGGAAQGGQGLGGQGGNKNKIESK SGEIKVSGSGEITSN-VNVTK GNKNKIESKGGAAQGGQGLGGQGGNK NKIESKGNKNKIESKGGAGQGGYGGLG GQGAGGGENKIESKGNENKIESKGGAG QGGYGGLGGQGAGGNKNKIESTGNKN KIESEGSGEIKVSGSGEITSNGNVNVTKK

[0394]

[0395] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0396] 4 Parallel pA56608 MEIETEGKVEVELEGEAEAKVKLEGK AEAKVKLEGKNNEAKVKLEGE- EIET-KVEVEL- solenoid NNEAKVKLEGEGGSGGGFEEIISSGPG GEAKVKLEGDKNEAKVKLEGK- AEAKVKLEGK- GYGPGGYGPGGYGPGGYGPGGYGPG NEAKVKLEGNKNEAKVKLEGD- EAKVKLEGE- GYGGEGKGEAKVKLEGDKNEAKVKL NEAKVKLEGNKNEAKVKLEGK- GEAKVKLEGD- EGKGGSGGGFEEIISSGPGGYGPGGYG NEAKVKLEGNKNEAKVKLEAD- EAKVKLEGK-NEAKVKLE- PGGYGPGGYGPGGYGPGGYGGEGENE SEAKVKLEGNKNEAKVK- EAKVKLEGD-NEAKVKLE- AKVKLEGNKNEAKVKLEGDGGSGGG VKLEGNKNEAKVK- EAKVKLEGK-NEAKVKLE- FEEIISSGPGGYGPGGYGPGGYGPGGY VKLEGGKNEAKVK- EAKVKLEAD-SEAKVKLE- GPGGYGPGGYGGEGENEAKVKLEGN VKLEGNKNEAKVK- EAKVK-VKLE-EAKVK- KNEAKVKLEGKGGSGGGFEEIISSGPG VKLEGNKNEAKVK- VKLE-EAKVK-VKLE- GYGPGGYGPGGYGPGGYGPGGYGPG VKLEGNKNEAKVK- EAKVK-VKLE-EAKVK- GYGGEGKNEAKVKLEGNKNEAKVKL VKLEGNNNEAKVK- VKLE-EAKVK-VKLE- EADGGSGGGFEEIISSGPGGYGPGGYG VKLEGNNNEAKVK- EAKVK-VKLE-EAKVK- PGGYGPGGYGPGGYGPGGYGGEGESE VKLEGNNQEAKVKLEGK- VKLE-EAKVKLEGK-ENE- AKVKLEGNKNEAKVKLEGNGGSGGG VKLEGNNQEAKVKLEGE- VKLEGN-EAKVKLEGE-KNE- FEEIISSGPGGYGPGGYGPGGYGPGGY VKLEGKNQEAKVKLEGK- VKLEGK-EAKVKLEGK- GPGGYGPGGYGGEGKSEAKVKLEGN VKLEGENNEAKVKLEGD- VKLEGE-EAKVKLEGD- KNEAKVKLEGNGGSGGGFEEIISSGPG VKLEGKNQEAKVKLE VKLEGK-EAKVKLE-VEL GYGPGGYGPGGYGPGGYGPGGYGPG GYGGEGESEAKVKLEGGKNEAKVKLE GNGGSGGGFEEIISSGPGGYGPGGYGP GGYGPGGYGPGGYGPGGYGGEGEAE AKVKLEGNKNEAKVKLEGNGGSGGG FEEIISSGPGGYGPGGYGPGGYGPGGY GPGGYGPGGYGGEGEAEAKVKLEGN KNEAKVKLEGNGGSGGGFEEIISSGPG GYGPGGYGPGGYGPGGYGPGGYGPG GYGGEGEAEAKVKLEGNKNEAKVKL EGNGGSGGGFEEIISSGPGGYGPGGYG PGGYGPGGYGPGGYGPGGYGGEGEAE AKVKLEGNNNEAKVKLEGNGGSGGG FEEIISSGPGGYGPGGYGPGGYGPGGY GPGGYGPGGYGGEGEAEAKVKLEGN NNEAKVKLEGNGGSGGGFEEIISSGPG GYGPGGYGPGGYGPGGYGPGGYGPG GYGGEGENEAKVKLEGNNQEAKVKL EGKGGSGGGFEEIISSGPGGYGPGGYG PGGYGPGGYGPGGYGPGGYGGEGENE

[0397]

[0398] AKVKLEGNNQEAKVKLEGEGGSGGGF

[0399] 59

[0400] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0401] EEIISSGPGGYGPGGYGPGGYGPGGYG PGGYGPGGYGGEGKNEAKVKLEGKN QEAKVKLEGKGGSGGGFEEIISSGPGG YGPGGYGPGGYGPGGYGPGGYGPGG YGGEGENEAKVKLEGENNEAKVKLEG DGGSGGGFEEIISSGPGGYGPGGYGPG GYGPGGYGPGGYGPGGYGGEGKGEA KVKLEGKNQEAKVKLEKGKKAEAEK KNEKVELK

[0402]

[0403] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0404] 5 Parallel pA56609 AEIKVKNSENLKIKSEGNTKLELESKG TKLELESKGENNKLELESKG- EIKV-LKIKS-TKLELES- solenoid ENNKLELESKGNGGYGPGPQGPGGPG NKLELESKGENNKLELESKG- KLELESKG-NKLELES- QQGPGSQGPGNNKLELESKGENNKLE NKLELESKGENNKLELESKG- KLELESKG-NKLELES- LESKGNGGYGPGPQGPGGPGQQGPGS NKLELESKGENNKLELESKG- KLELESKG-NKLELES- QGPGNNKLELESKGENNKLELESKGN NKLELESKGENNKLELESKA- KLELESKG-NKLELES- GGYGPGPQGPGGPGQQGPGSQGPGDN NKLELESKGENNKLELESKG- KLELESKA-NKLELES- KLELESKGENNKLELESKGNGGYGPG NKLELESKGEENKLELESKG- KLELESKG-NKLELES- PQGPGGPGQQGPGSQGPGENKLELESK TKLELESKGEKNKLELESKG- KLELESKG-TKLELES- GENNKLELESKANGGYGPGPQGPGGP TKLELESKGEENKLELESKG- KLELESKG-TKLELES- GQQGPGSQGPGENKLELESKGENNKL TKLELESKGEKNKLELESKG- KLELESKG-TKLELES- ELESKGNGGYGPGPQGPGGPGQQGPG SKLELESKGEKNKLELESKG- KLELESKG-SKLELES- SQGPGENKLELESKGEENKLELESKGD TKLELESKGEENKLELESKG- KLELESKG-TKLELES- GGYGPGPQGPGGPGQQGPGSQGPGET SKLELESKGEKNKLELESKA KLELESKG-SKLELES- KLELESKGEKNKLELESKGNGGYGPG KLELESKA-GSQ-SKLELES- PQGPGGPGQQGPGSQGPGETKLELESK KLELESKA GEENKLELESKGDGGYGPGPQGPGGP GQQGPGSQGPGKTKLELESKGEKNKL ELESKGDGGYGPGPQGPGGPGQQGPG SQGPGESKLELESKGEKNKLELESKGK GGYGPGPQGPGGPGQQGPGSQGPGNT KLELESKGEENKLELESKGDGGYGPGP QGPGGPGQQGPGSQGPGNSKLELESK GEKNKLELESKADGGYGPGPQGPGGP GQQGPGSQGPGNSKLELESKGDENKL

[0405]

[0406] ELESKADGTIKTKATEGGKGKAEV

[0407] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0408] 6 Parallel pA56610 AKIEIKNYENLEIESEGKTELKIELKGE ENELKIELKGNKNKAKAKSE- IEI-NLEI-TELKIEL- solenoid KNKAKAKSEGKSGPGGYGPGSQGPGS KNELKIELKGNKNKAKAK- KNKAKAKSE-ENELKIELK- QGPSGPGSQGPGGENELKIELKGNKNK KNELKIELKGNKNKAKAK- KNKAKAKSE-KNELKIELK- AKAKSEGESGPGGYGPGSQGPGSQGPS KTELKIELKGNKNKAKAK- KNKAKAK-KNELKIELK- GPGSQGPGGKNELKIELKGNKNKAKA KTELKIELKGNKNKAKAK- KNKAKAK-KTELKIELK- KSEGDSGPGGYGPGSQGPGSQGPSGPG KTELKIELKGNKNK- KNKAKAK-KTELKIELK- SQGPGGKNELKIELKGNKNKAKAKSE KTELKIELKGGKNK- KNKAKAK-KTELKIELK- GESGPGGYGPGSQGPGSQGPSGPGSQG KTELKIELKGNKNK- KNK-KTELKIELK-KNK- PGGKTELKIELKGNKNKAKAKSEADS KTELKIELKGNKNK- KTELKIELK-KNK- GPGGYGPGSQGPGSQGPSGPGSQGPG KSELKIELKGNKNK- KTELKIELK-KNK- GKTELKIELKGNKNKAKAKSEGDSGP NSELKIELKGNKNK- KSELKIELK-KNK-AKSE- GGYGPGSQGPGSQGPSGPGSQGPGGK NSELKIELKGNKNK NSELKIELK-KNK-AKSE- TELKIELKGNKNKAKAKSEGKSGPGG NSELKIELK-KNK-AKSE- YGPGSQGPGSQGPSGPGSQGPGGKTEL NSELKIELK-KNK-AKSEK KIELKGGKNKAKAKSEGDSGPGGYGP GSQGPGSQGPSGPGSQGPGGKTELKIE LKGNKNKAKAKSEGDSGPGGYGPGSQ GPGSQGPSGPGSQGPGGKTELKIELKG NKNKAKAKSEGDSGPGGYGPGSQGPG SQGPSGPGSQGPGGKSELKIELKGNKN KAKAKSEGDSGPGGYGPGSQGPGSQG PSGPGSQGPGGNSELKIELKGNKNKAK AKSEGKSGPGGYGPGSQGPGSQGPSGP GSQGPGGNSELKIELKGNKNKAKAKS EADSGPGGYGPGSQGPGSQGPSGPGSQ GPGGNSELKIELKGDKNKAKAKSEKS

[0409]

[0410] GKIEKEATEGAKVEAKV

[0411] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0412] 7 Parallel pA56612 AKIEIKNYENLEIESEGKTELKIELKGE SGKNKAKAKSEGKENELKIEL- KIEIK-LEIES-TELKIEL- solenoid GGLGGQGGYGGQGSQGAGQGGYGSG SGKNKAKAKSEGEKNELKIEL- SGKNKAKAKS-ENELKIEL- QGGSGKNKAKAKSEGKENELKIELKG SGKNKAKAKSEGDKNELKIEL- SGKNKAKAKS-KNELKIEL- NGGLGGQGGYGGQGSQGAGQGGYGS SGKNKAKAKSEGEKTELKIEL- SGKNKAKAKS-KNELKIEL- GQGGSGKNKAKAKSEGEKNELKIELK SGKNKAKAKSEADKTELKIEL- SGKNKAKAKS-KTELKIEL- GNGGLGGQGGYGGQGSQGAGQGGYG SGKNKAKAKSEGDKTELKIEL- SGKNKAKAKS-KTELKIEL- SGQGGSGKNKAKAKSEGDKNELKIEL SGKNKAKAKSEGKKTELKIEL- SGKNKAK-KTELKIEL- KGNGGLGGQGGYGGQGSQGAGQGGY SGKNKAKAKSEGDKTELKIEL- SGKNKAK-KTELKIEL- GSGQGGSGKNKAKAKSEGEKTELKIE SGKNKAKAKSEGDKTELKIEL- SGKNKAK-KTELKIEL- LKGNGGLGGQGGYGGQGSQGAGQGG SGKNKAKAKSEGDKSELKIEL- SGKNKAK-KTELKIEL- YGSGQGGSGKNKAKAKSEADKTELKI SGKNKAKAKSEGDNSELKIEL- SGKNKAK-KSELKIEL- ELKGNGGLGGQGGYGGQGSQGAGQG SGKNKAKAKSEGKNSELKIEL- SGKNKAK-NSELKIEL- GYGSGQGGSGKNKAKAKSEGDKTEL SGKNKAKAKSEADNSELKIEL SGKNKAK-NSELKIEL- KIELKGNGGLGGQGGYGGQGSQGAG SGKNKAK-NSELKIEL- QGGYGSGQGGSGKNKAKAKSEGKKT SGKNK-GKIE ELKIELKGGGGLGGQGGYGGQGSQGA GQGGYGSGQGGSGKNKAKAKSEGDK TELKIELKGNGGLGGQGGYGGQGSQG AGQGGYGSGQGGSGKNKAKAKSEGD KTELKIELKGNGGLGGQGGYGGQGSQ GAGQGGYGSGQGGSGKNKAKAKSEG DKSELKIELKGNGGLGGQGGYGGQGS QGAGQGGYGSGQGGSGKNKAKAKSE GDNSELKIELKGNGGLGGQGGYGGQG SQGAGQGGYGSGQGGSGKNKAKAKS EGKNSELKIELKGNGGLGGQGGYGGQ GSQGAGQGGYGSGQGGSGKNKAKAK SEADNSELKIELKGDGGLGGQGGYGG QGSQGAGQGGYGSGQGGSGKNKAKA

[0413]

[0414] KSEKSGKIEKEATEGAKVEAKV

[0415] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0416] 8 Parallel pA56613 AKIEIKNYENLEIESEGKTELKIELKGE KAKAKSEGKENELKIELKG- KIEIK-NLEIES-TELKIELKG- solenoid GGYGPGPQGPGGPGQQGPGSQGPGKN KAKAKSEGEKNELKIELKG- KAKAKS-ENELKIELKG- KAKAKSEGKENELKIELKGNGGYGPG KNKAKAKSEGDKNELKIELKG- KAKAKS-KNELKIELKG- PQGPGGPGQQGPGSQGPGKNKAKAKS KNKAKAKSEGEKTELKIELKG- KNKAKAKSE-KNELKIELKG- EGEKNELKIELKGNGGYGPGPQGPGGP KNKAKAKSEADKTELKIELKG- KNKAKAKSE-KTELKIELKG- GQQGPGSQGPGKNKAKAKSEGDKNEL KNKAKAKSEGDKTELKIELKG- KNKAKAKS-KTELKIELKG- KIELKGNGGYGPGPQGPGGPGQQGPG KNKAKAKSEGKKTELKIELKG- KNKAKAKS-KTELKIELKG- SQGPGKNKAKAKSEGEKTELKIELKG KNKAKAKSEGDKTELKIELKGN- KNKAKAKS-KTELKIELKG- NGGYGPGPQGPGGPGQQGPGSQGPGK KNKAKAKSEGDKTELKIELKGN- KNKAKAK-KTELKIELKGN- NKAKAKSEADKTELKIELKGNGGYGP KNKAKAKSEGDKSELKIELKGN- KNKAK-KTELKIELKGN- GPQGPGGPGQQGPGSQGPGKNKAKAK AKSEGKNSELKIELKGN- KNK-KSELKIELKGN- SEGDKTELKIELKGNGGYGPGPQGPGG KAKAKSEADNSELKIELKGD NSELKIELKGN-AKS- PGQQGPGSQGPGKNKAKAKSEGKKTE NSELKIELKGN-KAKAKS- LKIELKGGGGYGPGPQGPGGPGQQGP NSELKIELKGD-KAKAK- GSQGPGKNKAKAKSEGDKTELKIELK KIEKEA-KVEAK GNGGYGPGPQGPGGPGQQGPGSQGPG KNKAKAKSEGDKTELKIELKGNGGYG PGPQGPGGPGQQGPGSQGPGKNKAKA KSEGDKSELKIELKGNGGYGPGPQGPG GPGQQGPGSQGPGKNKAKAKSEGDNS ELKIELKGNGGYGPGPQGPGGPGQQG PGSQGPGKNKAKAKSEGKNSELKIELK GNGGYGPGPQGPGGPGQQGPGSQGPG KNKAKAKSEADNSELKIELKGDGGYG PGPQGPGGPGQQGPGSQGPGKNKAKA

[0417]

[0418] KSEKSGKIEKEATEGAKVEAKV

[0419] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0420] 9 Parallel pA56614 AKIEIKNYENLEIESEGKTELKIELKGES KNKAKAKSEGKENELKIELK- KIEI-NLEIES-TELKIELK- solenoid GPGGYGPGSQGPGSQGPSGPGSQGPG KNKAKAKSEGEKNELKIELK- KNKAKAKS-ENELKIELK- GKNKAKAKSEGKENELKIELKGNSGP KNKAKAKSEGDKNELKIELK- KNKAKAKS-KNELKIELK- GGYGPGSQGPGSQGPSGPGSQGPGGK KNKAKAKSEGEKTELKIELK- KNKAKAKS-KNELKIELK- NKAKAKSEGEKNELKIELKGNSGPGG KNKAKAKSEADKTELKIELK- KNKAKAK-KTELKIELK- YGPGSQGPGSQGPSGPGSQGPGGKNK KNKAKAKSEGDKTELKIELK- KNKAKAKS-KTELKIELK- AKAKSEGDKNELKIELKGNSGPGGYG KNKAKAKSEGKKTELKIELK- KNKAKAKS-KTELKIELK- PGSQGPGSQGPSGPGSQGPGGKNKAK KNKAKAKSEGDKTELKIELK- KNKAKAKS-KTELKIELK- AKSEGEKTELKIELKGNSGPGGYGPGS KNKAKAKSEGDKTELKIELK- KNKAKAKS-KTELKIELK- QGPGSQGPSGPGSQGPGGKNKAKAKS KNKAKAKSEGDKSELKIELK- KNKAKAK-KTELKIELK- EADKTELKIELKGNSGPGGYGPGSQGP KNKAKAKSEGDNSELKIELK- KNKAKAK-KSELKIELK- GSQGPSGPGSQGPGGKNKAKAKSEGD KNKAKAKSEGKNSELKIELK- KNKAK-NSELKIELK- KTELKIELKGNSGPGGYGPGSQGPGSQ NKAKAKSEADNSELKIELK KNKAK-NSELKIELK-NKAK- GPSGPGSQGPGGKNKAKAKSEGKKTE NSELKIELK-VEAK LKIELKGGSGPGGYGPGSQGPGSQGPS GPGSQGPGGKNKAKAKSEGDKTELKI ELKGNSGPGGYGPGSQGPGSQGPSGPG SQGPGGKNKAKAKSEGDKTELKIELK GNSGPGGYGPGSQGPGSQGPSGPGSQ GPGGKNKAKAKSEGDKSELKIELKGN SGPGGYGPGSQGPGSQGPSGPGSQGPG GKNKAKAKSEGDNSELKIELKGNSGP GGYGPGSQGPGSQGPSGPGSQGPGGK NKAKAKSEGKNSELKIELKGNSGPGG YGPGSQGPGSQGPSGPGSQGPGGKNK AKAKSEADNSELKIELKGDSGPGGYGP GSQGPGSQGPSGPGSQGPGGKNKAKA

[0421]

[0422] KSEKSGKIEKEATEGAKVEAKV

[0423] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0424] 10 Parallel pA56617 MEIETEGKVEVELEGEAEAKVKLEGK AEAKVKLEGKNNEAKVKLEGE- EIET-KVEVELE-AEAKVKLE- solenoid NNEAKVKLEGEGGLGGQGGYGGQGS SGKGEAKVKLEGDKNEAKVKLE NNEAKVKLEGE- QGAGQGGYGSGQGGSGKGEAKVKLE GK- SGKGEAKVKLE- GDKNEAKVKLEGKGGLGGQGGYGGQ SGENEAKVKLEGNKNEAKVKLE KNEAKVKLEGK- GSQGAGQGGYGSGQGGSGENEAKVK GD- SGENEAKVKLE- LEGNKNEAKVKLEGDGGLGGQGGYG SGENEAKVKLEGNKNEAKVKLE KNEAKVKLEGD- GQGSQGAGQGGYGSGQGGSGENEAK GK- SGENEAKVKLE- VKLEGNKNEAKVKLEGKGGLGGQGG SGKNEAKVKLEGNKNEAKVKLE KNEAKVKLEGK- YGGQGSQGAGQGGYGSGQGGSGKNE AD- SGKNEAKVKLE- AKVKLEGNKNEAKVKLEADGGLGGQ SGESEAKVKLEGNKNEAKVKLEG KNEAKVKLEAD- GGYGGQGSQGAGQGGYGSGQGGSGE N- SGESEAKVKLE- SEAKVKLEGNKNEAKVKLEGNGGLG SGKSEAKVKLEGNKNEAKVKLE KNEAKVKLEGN- GQGGYGGQGSQGAGQGGYGSGQGGS GN- SGKSEAKVKLEGNKNEAKV GKSEAKVKLEGNKNEAKVKLEGNGG SGESEAKVKLEGGKNEAKVKLEG KLEGN- LGGQGGYGGQGSQGAGQGGYGSGQG N- SGESEAKVKLEGGKNEAKV GSGESEAKVKLEGGKNEAKVKLEGNG SGEAEAKVKLEGNKNEAKVKLE KLEGN- GLGGQGGYGGQGSQGAGQGGYGSGQ GN- SGEAEAKVKLEGNKNEAKV GGSGEAEAKVKLEGNKNEAKVKLEG SGEAEAKVKLEGNKNEAKVKLE KLEGN-SGEAEAKVKLE- NGGLGGQGGYGGQGSQGAGQGGYGS GN- KNEAKVKLEGN- GQGGSGEAEAKVKLEGNKNEAKVKL SGEAEAKVKLEGNKNEAKVKLE SGEAEAKVKLE- EGNGGLGGQGGYGGQGSQGAGQGGY GN- EAKVKLEGN- GSGQGGSGEAEAKVKLEGNKNEAKV SGEAEAKVKLEGNNNEAKVKLE SGEAEAKVKLE- KLEGNGGLGGQGGYGGQGSQGAGQG GN- EAKVKLEGN- GYGSGQGGSGEAEAKVKLEGNNNEA SGEAEAKVKLEGNNNEAKVKLE SGEAEAKVKLE- KVKLEGNGGLGGQGGYGGQGSQGAG GN- NEAKVKLEGN- QGGYGSGQGGSGEAEAKVKLEGNNN SGENEAKVKLEGNNQEAKVKLE SGENEAKVKLE- EAKVKLEGNGGLGGQGGYGGQGSQG GK- QEAKVKLEGK- AGQGGYGSGQGGSGENEAKVKLEGN SGENEAKVKLEGNNQEAKVKLE SGENEAKVKLE- NQEAKVKLEGKGGLGGQGGYGGQGS GE- QEAKVKLEGE- QGAGQGGYGSGQGGSGENEAKVKLE SGKNEAKVKLEGKNQEAKVKLE SGKNEAKVKLEGKNQEAKV GNNQEAKVKLEGEGGLGGQGGYGGQ GK- KLEGK- GSQGAGQGGYGSGQGGSGKNEAKVK SGENEAKVKLEGENNEAKVKLEG SGENEAKVKLEGENNEAKV LEGKNQEAKVKLEGKGGLGGQGGYG D KLEGD-YGSG- GQGSQGAGQGGYGSGQGGSGENEAK SGKGEAKVKLEGK- VKLEGENNEAKVKLEGDGGLGGQGG EAKVKLEK-KAEAE-KVEL YGGQGSQGAGQGGYGSGQGGSGKGE AKVKLEGKNQEAKVKLEKGKKAEAE

[0425]

[0426] KKNEKVELK

[0427] 66

[0428] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0429] 11 Parallel pA56621 MEIETEGKVEVELEGEAEAKVKLEGK GNNEAKVKLEGEKGEAKVKLE- EIET-KVEVELE-AEAKVKL- solenoid GGYGPGPQGPGGPGQQGPGSQGPGNN GKNEAKVKLEGKENEAKVKLE- GNNEAKVKLE- EAKVKLEGEKGEAKVKLEGDGGYGP GKNEAKVKLEGDENEAKVKLE- KGEAKVKLE- GPQGPGGPGQQGPGSQGPGKNEAKVK GKNEAKVKLEGKKNEAKVKLE- GKNEAKVKLE- LEGKENEAKVKLEGNGGYGPGPQGPG GKNEAKVKLEADESEAKVKLE- ENEAKVKLE- GPGQQGPGSQGPGKNEAKVKLEGDEN GKNEAKVKLEGNKSEAKVKLE- GKNEAKVKLE- EAKVKLEGNGGYGPGPQGPGGPGQQG GKNEAKVKLEGNESEAKVKLE- ENEAKVKLE- PGSQGPGKNEAKVKLEGKKNEAKVKL GKNEAKVKLEGNEAEAKVKLE- GKNEAKVKLEGKKNEAKVK EGNGGYGPGPQGPGGPGQQGPGSQGP GKNEAKVKLEGNEAEAKVKLE- LE- GKNEAKVKLEADESEAKVKLEGNGG GKNEAKVKLEGNEAEAKVKLE- GKNEAKVKLEADESEAKVK YGPGPQGPGGPGQQGPGSQGPGKNEA GKNEAKVKLEGNEAEAKVKLE- LE-GKNEAKVKLE- KVKLEGNKSEAKVKLEGNGGYGPGPQ GNNEAKVKLEGNEAEAKVKLE- KSEAKVKLE- GPGGPGQQGPGSQGPGKNEAKVKLEG GNNEAKVKLEGNENEAKVKLE- GKNEAKVKLE- NESEAKVKLEGGGGYGPGPQGPGGPG GNQEAKVKLEGKENEAKVKLE- ESEAKVKLE- QQGPGSQGPGKNEAKVKLEGNEAEAK GNQEAKVKLEGEKNEAKVKLEG GKNEAKVKLE- VKLEGNGGYGPGPQGPGGPGQQGPGS K- EAEAKVKLE- QGPGKNEAKVKLEGNEAEAKVKLEG GNQEAKVKLEGKENEAKVKLEG GKNEAKVKLE- NGGYGPGPQGPGGPGQQGPGSQGPGK E- EAEAKVKLE- NEAKVKLEGNEAEAKVKLEGNGGYG GNNEAKVKLEGDKGEAKVKLEG GKNEAKVKLE- PGPQGPGGPGQQGPGSQGPGKNEAKV K EAEAKVKLE- KLEGNEAEAKVKLEGNGGYGPGPQGP GKNEAKVKLE- GGPGQQGPGSQGPGNNEAKVKLEGNE EAEAKVKLE- AEAKVKLEGNGGYGPGPQGPGGPGQQ GNNEAKVKLEGNEAEAKVK GPGSQGPGNNEAKVKLEGNENEAKVK LE- LEGNGGYGPGPQGPGGPGQQGPGSQG GNNEAKVKLEGNENEAKVK PGNQEAKVKLEGKENEAKVKLEGNG LE-GNQEAKVKLE- GYGPGPQGPGGPGQQGPGSQGPGNQE ENEAKVKLE- AKVKLEGEKNEAKVKLEGKGGYGPGP GNQEAKVKLE- QGPGGPGQQGPGSQGPGNQEAKVKLE KNEAKVKLEGK- GKENEAKVKLEGEGGYGPGPQGPGGP GNQEAKVKLE- GQQGPGSQGPGNNEAKVKLEGDKGE ENEAKVKLEGE- AKVKLEGKGGYGPGPQGPGGPGQQGP GNNEAKVKLE- GSQGPGNQEAKVKLEKGKKAEAEKK KGEAKVKLEGK-

[0430]

[0431] NEKVELK QEAKVKLE-KAEAEK-VEL

[0432] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0433] 12 Parallel pA56622 MEIETEGKVEVELEGEAEAKVKLEGKS NNEAKVKLEGEKGEAKVKL- EIETE-KVEVELE- solenoid GPGGYGPGSQGPGSQGPSGPGSQGPG KNEAKVKLEGKENEAKVKL- AEAKVKLE-GGY- GNNEAKVKLEGEKGEAKVKLEGDSGP KNEAKVKLEGDENEAKVKL- NNEAKVKLE-KGEAKVKL- GGYGPGSQGPGSQGPSGPGSQGPGGK KNEAKVKLEGKKNEAKVKL- KNEAKVKLE-ENEAKVKL- NEAKVKLEGKENEAKVKLEGNSGPGG KNEAKVKLEADESEAKVKL- KNEAKVKLE-ENEAKVKL- YGPGSQGPGSQGPSGPGSQGPGGKNE KNEAKVKLEGNKSEAKVKL- KNEAKVKLE-KNEAKVKL- AKVKLEGDENEAKVKLEGNSGPGGYG KNEAKVKLEGNESEAKVKL- KNEAKVKLE-ESEAKVKL- PGSQGPGSQGPSGPGSQGPGGKNEAK KNEAKVKLEGNEAEAKVKL- KNEAKVKLE-KSEAKVKL- VKLEGKKNEAKVKLEGNSGPGGYGPG KNEAKVKLEGNEAEAKVKL- KNEAKVKLE-ESEAKVKL- SQGPGSQGPSGPGSQGPGGKNEAKVK KNEAKVKLEGNEAEAKVKL- KNEAKVKLE-EAEAKVKL- LEADESEAKVKLEGNSGPGGYGPGSQ KNEAKVKLEGNEAEAKVKL- KNEAKVKLE-EAEAKVKL- GPGSQGPSGPGSQGPGGKNEAKVKLE NNEAKVKLEGNEAEAKVKL- KNEAKVKLE-EAEAKVKL- GNKSEAKVKLEGNSGPGGYGPGSQGP NNEAKVKLEGNENEAKVKL- KNEAKVKLEGNEAEAKVKL GSQGPSGPGSQGPGGKNEAKVKLEGN QEAKVKLEGKENEAKVKL- ESEAKVKLEGGSGPGGYGPGSQGPGS QEAKVKLEGEKNEAKVKL- NNEAKVKLEGNEAEAKVKL QGPSGPGSQGPGGKNEAKVKLEGNEA QEAKVKLEGKENEAKVKL- EAKVKLEGNSGPGGYGPGSQGPGSQG NEAKVKLEGDKGEAKVKL NNEAKVKLEGNENEAKVKL PSGPGSQGPGGKNEAKVKLEGNEAEA -QEAKVKLE-ENEAKVKL- KVKLEGNSGPGGYGPGSQGPGSQGPS QEAKVKLE-KNEAKVKL- GPGSQGPGGKNEAKVKLEGNEAEAKV QEAKVKLE-ENEAKVKL- KLEGNSGPGGYGPGSQGPGSQGPSGPG NEAKVKLEG-KGEAKVKL- SQGPGGKNEAKVKLEGNEAEAKVKLE QEAKVKLEK-EKVEL GNSGPGGYGPGSQGPGSQGPSGPGSQ GPGGNNEAKVKLEGNEAEAKVKLEG NSGPGGYGPGSQGPGSQGPSGPGSQGP GGNNEAKVKLEGNENEAKVKLEGNS GPGGYGPGSQGPGSQGPSGPGSQGPG GNQEAKVKLEGKENEAKVKLEGNSGP GGYGPGSQGPGSQGPSGPGSQGPGGN QEAKVKLEGEKNEAKVKLEGKSGPGG YGPGSQGPGSQGPSGPGSQGPGGNQE AKVKLEGKENEAKVKLEGESGPGGYG PGSQGPGSQGPSGPGSQGPGGNNEAK VKLEGDKGEAKVKLEGKSGPGGYGPG SQGPGSQGPSGPGSQGPGGNQEAKVK

[0434]

[0435] LEKGKKAEAEKKNEKVELK

[0436] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0437] 13 Parallel pA56624 MEIETEGKVEVELEGEAEAKVKLEGK GNNEAKVKLEGEKGEAKVKLE- EIET-KVEVELE-AEAKVKL- solenoid GGPGGSGPGQYGPGGAGPGQYGPGNN GKNEAKVKLEGKENEAKVKLE- GNNEAKVKLE- EAKVKLEGEKGEAKVKLEGDGGPGGS GKNEAKVKLEGDENEAKVKLE- KGEAKVKLE- GPGQYGPGGAGPGQYGPGKNEAKVK GKNEAKVKLEGKKNEAKVKLE- GKNEAKVKLE- LEGKENEAKVKLEGNGGPGGSGPGQY GKNEAKVKLEADESEAKVKLE- ENEAKVKLE- GPGGAGPGQYGPGKNEAKVKLEGDE GKNEAKVKLEGNKSEAKVKLE- GKNEAKVKLE- NEAKVKLEGNGGPGGSGPGQYGPGGA GKNEAKVKLEGNESEAKVKLE- ENEAKVKLE- GPGQYGPGKNEAKVKLEGKKNEAKV GKNEAKVKLEGNEAEAKVKLE- GKNEAKVKLE- KLEGNGGPGGSGPGQYGPGGAGPGQY GKNEAKVKLEGNEAEAKVKLE- KNEAKVKLE- GPGKNEAKVKLEADESEAKVKLEGNG GKNEAKVKLEGNEAEAKVKLE- GKNEAKVKLE- GPGGSGPGQYGPGGAGPGQYGPGKNE GKNEAKVKLEGNEAEAKVKLE- ESEAKVKLE- AKVKLEGNKSEAKVKLEGNGGPGGSG GNNEAKVKLEGNEAEAKVKLE- GKNEAKVKLE- PGQYGPGGAGPGQYGPGKNEAKVKLE GNNEAKVKLEGNENEAKVKLE- KSEAKVKLE- GNESEAKVKLEGGGGPGGSGPGQYGP GNQEAKVKLEGKENEAKVKLE- GKNEAKVKLE- GGAGPGQYGPGKNEAKVKLEGNEAE GNQEAKVKLEGEKNEAKVKLEG ESEAKVKLE- AKVKLEGNGGPGGSGPGQYGPGGAGP K- GKNEAKVKLE- GQYGPGKNEAKVKLEGNEAEAKVKL GNQEAKVKLEGKENEAKVKLEG EAEAKVKLE- EGNGGPGGSGPGQYGPGGAGPGQYGP E- GKNEAKVKLE- GKNEAKVKLEGNEAEAKVKLEGNGG GNNEAKVKLEGDKGEAKVKLEG EAEAKVKLE- PGGSGPGQYGPGGAGPGQYGPGKNEA K GKNEAKVKLE- KVKLEGNEAEAKVKLEGNGGPGGSGP EAEAKVKLE- GQYGPGGAGPGQYGPGNNEAKVKLE GKNEAKVKLE- GNEAEAKVKLEGNGGPGGSGPGQYGP EAEAKVKLE- GGAGPGQYGPGNNEAKVKLEGNENE GNNEAKVKLE- AKVKLEGNGGPGGSGPGQYGPGGAGP EAEAKVKLE- GQYGPGNQEAKVKLEGKENEAKVKL GNNEAKVKLE- EGNGGPGGSGPGQYGPGGAGPGQYGP ENEAKVKLE- GNQEAKVKLEGEKNEAKVKLEGKGG GNQEAKVKLE- PGGSGPGQYGPGGAGPGQYGPGNQEA ENEAKVKLE- KVKLEGKENEAKVKLEGEGGPGGSGP GNQEAKVKLE- GQYGPGGAGPGQYGPGNNEAKVKLE KNEAKVKLEGK- GDKGEAKVKLEGKGGPGGSGPGQYGP GNQEAKVKLE- GGAGPGQYGPGNQEAKVKLEKGKKA ENEAKVKLEGE- EAEKKNEKVELK GNNEAKVKLE- KGEAKVKLEGK-GQY- GNQEAKVKLE-KAEAEK-

[0438]

[0439] VEL

[0440] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0441] 14 Parallel pA56628 MVKVEGEGVKVKVEGEAKAEAKAEG KIEIKLEGKKNKAEAKAEGD- VKVE-VKVKVE- solenoid EGGPGGSGPGQYGPGGAGPGQYGPGG KIEIKLEGEENKAEAKAEGE- AKAEAKAEGE-KIEIKLE- NKIEIKLEGKKNKAEAKAEGDGGPGG KIEIKLEGDKNKAEAKAEGEG- KNKAEAKAEGD-KIEIKLE- SGPGQYGPGGAGPGQYGPGGNKIEIKL KIEIKLEGEKNKAEAKAEGEG- ENKAEAKAEGE-KIEIKLE- EGEENKAEAKAEGEGGPGGSGPGQYG KIEIKLEGDEAKAEAKAEGEG KAEAKAEGEG-KIEIKLE- PGGAGPGQYGPGGNKIEIKLEGDKNK KAEAKAEGEG-KIEIKLE- AEAKAEGEGGPGGSGPGQYGPGGAGP AKAEAKAEGEG-KIEIKLE- GQYGPGGNKIEIKLEGEKNKAEAKAE GKAKAL-TEVD GEGGPGGSGPGQYGPGGAGPGQYGPG GNKIEIKLEGDEAKAEAKAEGEGGPGG SGPGQYGPGGAGPGQYGPGGNKIEIKL EGSGKAKALPGETEVDDEGAKVKVEE KK

[0442] 16 Parallel pA56631 MVKVEGEGVKVKVEGEAKAEAKAEG KNKAEAKAEGDGNKIEIKLEG- VKVE-KVKVE-KAEAKAE- solenoid EGNKIEIKLEGKGGYGPGPQGPGGPGQ ENKAEAKAEGEGNKIEIKLEG- KIEIKLEG-KNKAEAKAE- QGPGSQGPGKNKAEAKAEGDGNKIEI KNKAEAKAEGEGNKIEIKLEG- KIEIKLEG-ENKAEAKAE- KLEGEGGYGPGPQGPGGPGQQGPGSQ KNKAEAKAEGEGNKIEIKLE KIEIKLEG-KNKAEAKAE- GPGENKAEAKAEGEGNKIEIKLEGDGG KIEIKLEG-KNKAEAKAE- YGPGPQGPGGPGQQGPGSQGPGKNKA KIEIKLE-GEAKAEAKAE- EAKAEGEGNKIEIKLEGEGGYGPGPQG KIEIKLE-KVKVEEK PGGPGQQGPGSQGPGKNKAEAKAEGE GNKIEIKLEGDGGYGPGPQGPGGPGQQ GPGSQGPGEAKAEAKAEGEGNKIEIKL EGSGKAKALPGETEVDDEGAKVKVEE KK

[0443] 17 Parallel pA56633 MMKIKKTENAEVKATGEGGSGGQGG GNNEIEVESEKGKNKAELKGE- MKIKK-AEVKATGE- solenoid YGGLGSQGAGQGGYGAGQGGAGNNE GKNKAELKGNNNKAELKGN- GNNEIEVES-KNKAELKGE- IEVESEKGKNKAELKGEGGSGGQGGY GKNKAELKGNNNKAELKGN- GKNKAELK-NNKAELKGN- GGLGSQGAGQGGYGAGQGGAGKNKA GKNKAELKGNDNKAELKGN- GKNKAELK-NNKAELKGN- ELKGNNNKAELKGNGGSGGQGGYGG GKNKAELKGNKNKAELKGN- GKNKAELK-DNKAELKGN- LGSQGAGQGGYGAGQGGAGKNKAEL GKNKAELKGNKNKAELKGN- GKNKAELK-KNKAELKGN- KGNNNKAELKGNGGSGGQGGYGGLG GKNKAELKGNKNKAELKGN- GKNKAELK-KNKAELKGN- SQGAGQGGYGAGQGGAGKNKAELKG GKNKAELKGNKNKAELKGG- GKNKAELK-KNKAELKGN- NDNKAELKGNGGSGGQGGYGGLGSQ GENKAELKGKKNKAELKGE- GKNKAELK-KNKAELKGG- GAGQGGYGAGQGGAGKNKAELKGN GKNKAELKGNKNKAELTGK GENKAELK-KNKAELKGE- KNKAELKGNGGSGGQGGYGGLGSQG GKNKAELK-KNKAELTGK- AGQGGYGAGQGGAGKNKAELKGNK GTTEVTIK-GEAYVK-EGKVE NKAELKGNGGSGGQGGYGGLGSQGA

[0444]

[0445] GQGGYGAGQGGAGKNKAELKGNKN

[0446] 70

[0447] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0448] KAELKGNGGSGGQGGYGGLGSQGAG QGGYGAGQGGAGKNKAELKGNKNK AELKGGGGSGGQGGYGGLGSQGAGQ GGYGAGQGGAGENKAELKGKKNKAE LKGEGGSGGQGGYGGLGSQGAGQGG YGAGQGGAGKNKAELKGNKNKAELT GKGGSGGQGGYGGLGSQGAGQGGYG AGQGGAGTTEVTIKGGGEAYVKGEGK VEV

[0449] 18 Parallel pA56634 MMKIKKTENAEVKATGEGGLGGQGG GNNEIEVESEKGKNKAELKGE- MKIK-AEVKATGE- solenoid YGGQGSQGAGQGGYGSGQGGSGNNEI GKNKAELKGNNNKAELKGN- GNNEIEVE-NKAELKGE- EVESEKGKNKAELKGEGGLGGQGGY GKNKAELKGNNNKAELK- GKNKAELK-KAELKGN- GGQGSQGAGQGGYGSGQGGSGKNKA GKNKAELKGNDNKAEL- GKNKAELK-KAELK- ELKGNNNKAELKGNGGLGGQGGYGG GKNKAELKGNKNKAEL- GKNKAELK-KAEL- QGSQGAGQGGYGSGQGGSGKNKAEL GKNKAELKGNKNKAEL- GKNKAELK-KAEL- KGNNNKAELKGNGGLGGQGGYGGQG GKNKAELKGNKNKAEL- GKNKAELK-KAEL- SQGAGQGGYGSGQGGSGKNKAELKG GKNKAELKGNKNKAELKGG- GKNKAELK-KAEL- NDNKAELKGNGGLGGQGGYGGQGSQ GENKAELKGKKNKAELKGE- GKNKAELK-KAELKGG- GAGQGGYGSGQGGSGKNKAELKGNK GKNKAELKGNKNKAELTGK GENKAELK-KAELKGE- NKAELKGNGGLGGQGGYGGQGSQGA GKNKAELK-NKAELTGK- GQGGYGSGQGGSGKNKAELKGNKNK GTTEVTIK-GEAYVK-EGKVE AELKGNGGLGGQGGYGGQGSQGAGQ GGYGSGQGGSGKNKAELKGNKNKAE LKGNGGLGGQGGYGGQGSQGAGQGG YGSGQGGSGKNKAELKGNKNKAELK GGGGLGGQGGYGGQGSQGAGQGGYG SGQGGSGENKAELKGKKNKAELKGEG GLGGQGGYGGQGSQGAGQGGYGSGQ GGSGKNKAELKGNKNKAELTGKGGL GGQGGYGGQGSQGAGQGGYGSGQGG

[0450]

[0451] SGTTEVTIKGGGEAYVKGEGKVEV

[0452] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0453] 19 Parallel pA56635 MMKIKKTENAEVKATGEGGYGPGPQ NNEIEVESEKGKNKAELKG- MKIK-EVKATG-NNEIEVE- solenoid GPGGPGQQGPGSQGPGNNEIEVESEKG KNKAELKGNNNKAELKG- KAELKG-KNKAELK- KNKAELKGEGGYGPGPQGPGGPGQQG KNKAELKGNNNKAEL- KAELKG-KNKAELK-KAEL- PGSQGPGKNKAELKGNNNKAELKGN KNKAELKGNDNKAEL- KNKAELK-KAEL- GGYGPGPQGPGGPGQQGPGSQGPGKN KNKAELKGNKNKAEL- KNKAELK-KAEL- KAELKGNNNKAELKGNGGYGPGPQG KNKAELKGNKNKAEL- KNKAELK-KAEL- PGGPGQQGPGSQGPGKNKAELKGNDN GKNKAELKGNKNKAEL- GKNKAELK-KAEL- KAELKGNGGYGPGPQGPGGPGQQGPG GKNKAELKGNKNKAEL- GKNKAELK-KAEL- SQGPGKNKAELKGNKNKAELKGNGG GENKAELKGKKNKAELKGE- GENKAELK-KAELKGE- YGPGPQGPGGPGQQGPGSQGPGKNKA KNKAELKGNKNKAELTG KNKAELK-NKAELTG- ELKGNKNKAELKGNGGYGPGPQGPG TTEVTIK-GEAYVK-EGKVE GPGQQGPGSQGPGKNKAELKGNKNK AELKGNGGYGPGPQGPGGPGQQGPGS QGPGKNKAELKGNKNKAELKGGGGY GPGPQGPGGPGQQGPGSQGPGENKAE LKGKKNKAELKGEGGYGPGPQGPGGP GQQGPGSQGPGKNKAELKGNKNKAE LTGKGGYGPGPQGPGGPGQQGPGSQG PGTTEVTIKGGGEAYVKGEGKVEV

[0454] 20 Parallel pA56636 MMKIKKTENAEVKATGESGPGGYGPG NNEIEVESEKGKNKAEL- MKIKKT-AEVKA- solenoid SQGPGSQGPSGPGSQGPGGNNEIEVES KNKAELKGNNNKAEL- NNEIEVES-KNKAEL- EKGKNKAELKGESGPGGYGPGSQGPG KNKAELKGNNNKAEL- KNKAELK-NNKAEL- SQGPSGPGSQGPGGKNKAELKGNNNK KNKAELKGNDNKAEL- KNKAELK-NNKAEL- AELKGNSGPGGYGPGSQGPGSQGPSGP KNKAELKGNKNKAEL- KNKAELK-DNKAEL- GSQGPGGKNKAELKGNNNKAELKGN KNKAELKGNKNKAEL- KNKAELK-KNKAEL- SGPGGYGPGSQGPGSQGPSGPGSQGPG KNKAELKGNKNKAELK- KNKAELK-KNKAEL- GKNKAELKGNDNKAELKGNSGPGGY KNKAELKGNKNKAELKGG- KNKAELK-KNKAELK- GPGSQGPGSQGPSGPGSQGPGGKNKA ENKAELKGKKNKAELKGE- KNKAELK-KNKAELKGG- ELKGNKNKAELKGNSGPGGYGPGSQG KNKAELKGNKNKAELTGK ENKAELK-KNKAELKGE- PGSQGPSGPGSQGPGGKNKAELKGNK KNKAELK-KNKAELTGK- NKAELKGNSGPGGYGPGSQGPGSQGP TTEVTIK-GEAYV-GKVE SGPGSQGPGGKNKAELKGNKNKAELK GNSGPGGYGPGSQGPGSQGPSGPGSQ GPGGKNKAELKGNKNKAELKGGSGP GGYGPGSQGPGSQGPSGPGSQGPGGE NKAELKGKKNKAELKGESGPGGYGPG SQGPGSQGPSGPGSQGPGGKNKAELK

[0455]

[0456] GNKNKAELTGKSGPGGYGPGSQGPGS

[0457] 72

[0458] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0459] QGPSGPGSQGPGGTTEVTIKGGGEAYV KGEGKVEV

[0460] 21 Parallel pA56637 MMKIKKTENAEVKATGEGPGQQGPG NNEIEVESEKGKNKAELKG- MKIKKT-AEVKATG- solenoid GYGPGQQGPSGPGNNEIEVESEKGKN KNKAELKGNNNKAELKG- NNEIEVES-NKAELKG- KAELKGEGPGQQGPGGYGPGQQGPSG KNKAELKGNNNKAELKG- KNKAELK-KAELKG- PGKNKAELKGNNNKAELKGNGPGQQ KNKAELKGNDNKAELKG- KNKAELK-KAELKG- GPGGYGPGQQGPSGPGKNKAELKGNN KNKAELKGNKNKAELKG- KNKAELK-KAELKG- NKAELKGNGPGQQGPGGYGPGQQGPS KNKAELKGNKNKAELKGN- KNKAELK-KAELKG- GPGKNKAELKGNDNKAELKGNGPGQ KNKAELKGNKNKAELKGN- KNKAELK-KAELKGN- QGPGGYGPGQQGPSGPGKNKAELKGN KNKAELKGNKNKAELKG- KNKAELK-KAELKGN- KNKAELKGNGPGQQGPGGYGPGQQG ENKAELKGKKNKAELKG- KNKAELK-KAELKG- PSGPGKNKAELKGNKNKAELKGNGPG KNKAELKGNKNKAELTG ENKAELK-KAELKG- QQGPGGYGPGQQGPSGPGKNKAELKG KNKAELK-NKAELTG- NKNKAELKGNGPGQQGPGGYGPGQQ TTEVTIK-GEAYVK-EGKVE GPSGPGKNKAELKGNKNKAELKGGGP GQQGPGGYGPGQQGPSGPGENKAELK GKKNKAELKGEGPGQQGPGGYGPGQ QGPSGPGKNKAELKGNKNKAELTGKG PGQQGPGGYGPGQQGPSGPGTTEVTIK

[0461]

[0462] GGGEAYVKGEGKVEV

[0463] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0464] 22 Parallel pA56638 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAELKG- MKIK-AEVKA-NNEIEVES- solenoid KGKNKAELKGEKNKAELKGNGGLGG GNNKAELKGNKNKAEL- KNKAELK-KNKAELKG- QGGYGGQGSQGAGQGGYGSGQGGSG GNNKAELKGNKNKAEL- GNNKAELK-KAEL- NNKAELKGNKNKAELKGNGGLGGQG GDNKAELKGNKNKAEL- GNNKAELK-KAEL- GYGGQGSQGAGQGGYGSGQGGSGNN GKNKAELKGNKNKAEL- GDNKAELK-KAEL- KAELKGNKNKAELKGNGGLGGQGGY GKNKAELKGNKNKAEL- GKNKAELK-KAEL- GGQGSQGAGQGGYGSGQGGSGDNKA GKNKAELKGNKNKAEL- GKNKAELK-KAEL- ELKGNKNKAELKGNGGLGGQGGYGG GKNKAELKGGENKAEL- GKNKAELK-NKAEL- QGSQGAGQGGYGSGQGGSGKNKAEL GKNKAELKGEKNKAEL GKNKAELK-ENKAEL- KGNKNKAELKGNGGLGGQGGYGGQG GKNKAELK-KNKAEL- SQGAGQGGYGSGQGGSGKNKAELKG NKAELTGKTTEVTI- NKNKAELKGNGGLGGQGGYGGQGSQ GEAYVKGEGKVE GAGQGGYGSGQGGSGKNKAELKGNK NKAELKGNGGLGGQGGYGGQGSQGA GQGGYGSGQGGSGKNKAELKGGENK AELKGKGGLGGQGGYGGQGSQGAGQ GGYGSGQGGSGKNKAELKGEKNKAE LKGNGGLGGQGGYGGQGSQGAGQGG YGSGQGGSGKNKAELTGKTTEVTIKG GGEAYVKGEGKVEV

[0465] 23 Parallel pA56639 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAELKG- MKIKKT-AEVKA- solenoid KGKNKAELKGEKNKAELKGNGGYGP GNNKAELKGNKNKAELK- NNEIEVES-KNKAELK- GPQGPGGPGQQGPGSQGPGNNKAELK GNNKAELKGNKNKAEL- KNKAELKG-GNNKAELK- GNKNKAELKGNGGYGPGPQGPGGPG GDNKAELKGNKNKAEL- KNKAELK-GNNKAELK- QQGPGSQGPGNNKAELKGNKNKAEL GKNKAELKGNKNKAEL- KNKAEL-GDNKAELK- KGNGGYGPGPQGPGGPGQQGPGSQGP GKNKAELKGNKNKAEL- KNKAEL-GKNKAELK- GDNKAELKGNKNKAELKGNGGYGPG KNKAELKGNKNKAEL- KNKAEL-GKNKAELK- PQGPGGPGQQGPGSQGPGKNKAELKG KNKAELKGGENKAEL- KNKAEL- NKNKAELKGNGGYGPGPQGPGGPGQ KNKAELKGEKNKAEL KNKAELKGNKNKAEL- QGPGSQGPGKNKAELKGNKNKAELK KNKAELKGGENKAEL- GNGGYGPGPQGPGGPGQQGPGSQGPG KNKAELKGEKNKAEL- KNKAELKGNKNKAELKGNGGYGPGP KNKAELTGKTTEVTI- QGPGGPGQQGPGSQGPGKNKAELKGG GEAYVKGEGKVE ENKAELKGKGGYGPGPQGPGGPGQQG PGSQGPGKNKAELKGEKNKAELKGNG GYGPGPQGPGGPGQQGPGSQGPGKNK AELTGKTTEVTIKGGGEAYVKGEGKV

[0466]

[0467] EV

[0468] 74

[0469] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0470] 24 Parallel pA56640 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAEL- MKIKKTENAEVKA- solenoid KGKNKAELKGEKNKAELKGNSGPGG NNKAELKGNKNKAEL- NNEIEVESEKGK-KNKAELK- YGPGSQGPGSQGPSGPGSQGPGGNNK NNKAELKGNKNKAEL- KNKAEL-NNKAELK- AELKGNKNKAELKGNSGPGGYGPGSQ DNKAELKGNKNKAEL- KNKAEL-NNKAELK- GPGSQGPSGPGSQGPGGNNKAELKGN KNKAELKGNKNKAEL- KNKAEL-DNKAELK- KNKAELKGNSGPGGYGPGSQGPGSQG KNKAELKGNKNKAEL- KNKAEL-KNKAELK- PSGPGSQGPGGDNKAELKGNKNKAEL KNKAELKGNKNKAEL- KNKAEL-KNKAELK- KGNSGPGGYGPGSQGPGSQGPSGPGS KNKAELKGGENKAEL- KNKAEL-KNKAELK- QGPGGKNKAELKGNKNKAELKGNSG KNKAELKGEKNKAEL KNKAEL-KNKAELK- PGGYGPGSQGPGSQGPSGPGSQGPGG ENKAEL-KNKAELK- KNKAELKGNKNKAELKGNSGPGGYG KNKAEL- PGSQGPGSQGPSGPGSQGPGGKNKAEL KNKAELTGKTTEVTI- KGNKNKAELKGNSGPGGYGPGSQGPG GEAYVKGEGKVE SQGPSGPGSQGPGGKNKAELKGGENK AELKGKSGPGGYGPGSQGPGSQGPSGP GSQGPGGKNKAELKGEKNKAELKGNS GPGGYGPGSQGPGSQGPSGPGSQGPG GKNKAELTGKTTEVTIKGGGEAYVKG EGKVEV

[0471] 25 Parallel pA56641 MMKIKKTENAEVKATGENNEIEVESE GKNKAELKGEKNKAELKG- MKIKK-AEVKA-NNEIEVES- solenoid KGKNKAELKGEKNKAELKGNGPGQQ PGNNKAELKGNKNKAELK- GKNKAELK-KNKAELKG- GPGGYGPGQQGPSGPGNNKAELKGNK NKAELKGNKNKAELK- PGNNKAELK-KNKAELK- NKAELKGNGPGQQGPGGYGPGQQGPS NKAELKGNKNKAELK- NKAELK-KNKAELK- GPGNNKAELKGNKNKAELKGNGPGQ NKAELKGNKNKAELK- NKAELK-KNKAELK- QGPGGYGPGQQGPSGPGDNKAELKGN NKAELKGNKNKAELK- NKAELK-KNKAELK- KNKAELKGNGPGQQGPGGYGPGQQG NKAELKGNKNKAELK- NKAELK-KNKAELK- PSGPGKNKAELKGNKNKAELKGNGPG NKAELKGGENKAELK- NKAELK-KNKAELK- QQGPGGYGPGQQGPSGPGKNKAELKG NKAELKGEKNKAELK NKAELK-ENKAELK- NKNKAELKGNGPGQQGPGGYGPGQQ NKAELK-KNKAELK- GPSGPGKNKAELKGNKNKAELKGNGP NKAELTGKTTEVTIK- GQQGPGGYGPGQQGPSGPGKNKAELK GEAYVKGEGKVE GGENKAELKGKGPGQQGPGGYGPGQ QGPSGPGKNKAELKGEKNKAELKGNG PGQQGPGGYGPGQQGPSGPGKNKAEL

[0472]

[0473] TGKTTEVTIKGGGEAYVKGEGKVEV

[0474] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0475] 26 Parallel pA56642 MMKIKKTENAEVKATGENNEIEVESE NKAELKGEKNKAELKGN- MKIKK-AEVKA-NNEIEVES- solenoid KGKNKAELKGEKNKAELKGNGGPGG NKAELKGNKNKAELKGN- NKAELK-KNKAELKGN- SGPGQYGPGGAGPGQYGPGNNKAELK NKAELKGNKNKAELKGN- NKAELK-KNKAELKGN- GNKNKAELKGNGGPGGSGPGQYGPG NKAELKGNKNKAEL- NKAELK-KNKAELKGN- GAGPGQYGPGNNKAELKGNKNKAEL NKAELKGNKNKAEL- NKAELK-KNKAEL- KGNGGPGGSGPGQYGPGGAGPGQYGP KNKAELKGNKNKAEL- NKAELK-KNKAEL- GDNKAELKGNKNKAELKGNGGPGGS KNKAELKGNKNKAEL- KNKAELK-KNKAEL- GPGQYGPGGAGPGQYGPGKNKAELK KNKAELKGGENKAEL- KNKAELKGNKNKAEL- GNKNKAELKGNGGPGGSGPGQYGPG KNKAELKGEKNKAEL KNKAELKGGENKAEL- GAGPGQYGPGKNKAELKGNKNKAEL KNKAELKGEKNKAEL- KGNGGPGGSGPGQYGPGGAGPGQYGP KNKAELT-TTEVTI- GKNKAELKGNKNKAELKGNGGPGGS GEAYVK-GKVE GPGQYGPGGAGPGQYGPGKNKAELK GGENKAELKGKGGPGGSGPGQYGPGG AGPGQYGPGKNKAELKGEKNKAELK GNGGPGGSGPGQYGPGGAGPGQYGPG KNKAELTGKTTEVTIKGGGEAYVKGE GKVEV

[0476] 27 Parallel pA56653 AKIEIKNYENLEIESEGKTELKIELKGE KAKAKSEGKENELKIELK- KIEI-NLEIES-TELKIELK- solenoid GGAGQGGYGGLGGQGAGRGAGKNK KAKAKSEGEKNELKIELK- KAKAKS-ENELKIELK- AKAKSEGKENELKIELKGNGQGGYGG KAKAKSEGDKNELKIELK- KAKAKS-KNELKIELK- LGGQGAGRGAGKNKAKAKSEGEKNE KAKAKSEGEKTELKIELK- KAKAKS-KNELKIELK- LKIELKGNGGAGQGGYGGLGSQGAGR KAKAKSEADKTELKIELK- KAKAKS-KTELKIELK- GGYGGQGAGKNKAKAKSEGDKNELK KAKAKSEGDKTELKIELK- KAKAKS-KTELKIELK- IELKGNQGGYGDLGSQGAGKNKAKA KAKAKSEGKKTELKIELK- KAKAKS-KTELKIELK- KSEGEKTELKIELKGNGGAGQGGYGG KAKAKSEGDKTELKIELK- KAKAKS-KTELKIELK- LGGQGAGQGAGKNKAKAKSEADKTE KAKAKSEGDKTELKIELK- KAKAK-KTELKIELK-KAK- LKIELKGNGQGGYGGLGGQGAGRGA KAKAKSEGDKSELKIELK- KTELKIELK-KAK- GKNKAKAKSEGDKTELKIELKGNGGA KAKAKSEGDNSELKIELK- KSELKIELK-KAK- AQGGQGLGGQGKNKAKAKSEGKKTE KAKAKSEGKNSELKIELK- NSELKIELK-KAK- LKIELKGGGGAGQGGYGGLGGQGAG KAKAKSEADNSELKIELK NSELKIELK-KAK- KNKAKAKSEGDKTELKIELKGNGGAA NSELKIELK-KAK-VEAK QGGQGLGGQGKNKAKAKSEGDKTEL KIELKGNGGAGQGGYGGLGSQGAGR GGYGGQGAGKNKAKAKSEGDKSELKI ELKGNGGAGQGGYGGLGSQGAGRGA KNKAKAKSEGDNSELKIELKGNGGAG QGGYGGLGSQGAGRGGYGGQGAGKN

[0477]

[0478] KAKAKSEGKNSELKIELKGNGGAGQG

[0479] 76

[0480] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0481] GYGGLGGQGAGQGAGKNKAKAKSEA DNSELKIELKGDGGAGQGGYGGLGSQ GAGRGGYGGQGAGKNKAKAKSEKSG KIEKEATEGAKVEAKV

[0482] 28 Parallel pA56654 MEIETEGKVEVELEGEAEAKVKLEGK AEAKVKLEGKNNEAKVKLEGE- EIET-KVEVELE-AEAKVKL- solenoid NNEAKVKLEGEGQGGYGGLGGQGAG KGEAKVKLEGDKNEAKVKLEGK- NNEAKVKLEGE- RGAGKGEAKVKLEGDKNEAKVKLEG ENEAKVKLEGNKNEAKVKLEG- KGEAKVKLE- KGGAGQGGYGGLGGQGAGRGAGENE ENEAKVKLEGNKNEAKVKLEG- KNEAKVKLEGK- AKVKLEGNKNEAKVKLEGDGGAGQG KNEAKVKLEGNKNEAKVKLEA- ENEAKVKLE- GYGGLGSQGAGRGGYGGQGAENEAK ESEAKVKLEGNKNEAKVKLEG- KNEAKVKLEG- VKLEGNKNEAKVKLEGKGGAGQGGY KSEAKVKLEGNKNEAKVKLEGN- ENEAKVKLE- GGLGGQGAGRGAGKNEAKVKLEGNK ESEAKVKLEGGKNEAKVKLEGN- KNEAKVKLEG- NEAKVKLEADGGAGQGGYGGLGGQG EAEAKVKLEGNKNEAKVKLEG- KNEAKVKLE- AGESEAKVKLEGNKNEAKVKLEGNG GEAEAKVKLEGNKNEAKVKLEG- KNEAKVKLEA- GAGQGGYGGLGGQGAGQGAGKSEAK GEAEAKVKLEGNKNEAKVKLEG ESEAKVKLE- VKLEGNKNEAKVKLEGNGGAGQGGY N- KNEAKVKLEG- GGLGSQGAGRGAESEAKVKLEGGKNE EAEAKVKLEGNNNEAKVKLEGN- KSEAKVKLEGNKNEAKVKL AKVKLEGNQGGYGDLGSQGAGEAEA EAEAKVKLEGNNNEAKVKLEGN- EGN- KVKLEGNKNEAKVKLEGNGGAGQGG ENEAKVKLEGNNQEAKVKLEGK- ESEAKVKLEGGKNEAKVKL YGGLGGQGAGRGAGEAEAKVKLEGN ENEAKVKLEGNNQEAKVKLEGE- EGN- KNEAKVKLEGNGGAGQGGYGGLGGQ KNEAKVKLEGKNQEAKVKLEGK- EAEAKVKLEGNKNEAKVKL GAGRGAGEAEAKVKLEGNKNEAKVK ENEAKVKLEGENNEAKVKLEG EG-GEAEAKVKLE- LEGNGGAGQGGYGGLGSQGAGRGGY KNEAKVKLEG- GGQGAEAEAKVKLEGNNNEAKVKLE GEAEAKVKLE- GNGGAGQGGYGGLGGQGAGRGAGEA EAKVKLEGN-EAEAKVKLE- EAKVKLEGNNNEAKVKLEGNGGAGQ EAKVKLEGN-EAEAKVKLE- GGYGGLGSQGAGRGAENEAKVKLEG NEAKVKLEGN-

[0483]

[0484] NNQEAKVKLEGKGGAGQGGYGGLGS ENEAKVKLE- 77

[0485] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0486] QGAGRGAENEAKVKLEGNNQEAKVK QEAKVKLEGK- LEGEGGAGQGGYGGLGSQGAGRGGY ENEAKVKLE- GGQGAKNEAKVKLEGKNQEAKVKLE QEAKVKLEGE- GKGGAAQGGQGLGGQGENEAKVKLE KNEAKVKLE- GENNEAKVKLEGDGGAGQGGYGGLG QEAKVKLEGK- SQGAGRGGYGGQGAGKGEAKVKLEG ENEAKVKLEGE- KNQEAKVKLEKGKKAEAEKKNEKVE EAKVKLEG- LK KGEAKVKLEGK- EAKVKLEK

[0487] 29 Parallel pA56657 MEIETEGKVEVELEGEAEAKVKLEGK AEAKVKLEGKNNEAKVKLEGEG EIET-KVEVELE-AEAKVKL- solenoid NNEAKVKLEGEGGAGQGGYGGLGGQ GAG- NNEAKVKLEGEGGAG- GAGRGAGKGEAKVKLEGDKNEAKVK GKGEAKVKLEGDKNEAKVKLEG GKGEAKVKLEGDKNEAKVK LEGKGQGGYGGLGGQGAGRGAGENE KGQG- LEGKGQG- AKVKLEGNKNEAKVKLEGDQGGYGD GENEAKVKLEGNKNEAKVKLEG GENEAKVKLEGNKNEAKVK LGSQGAGENEAKVKLEGNKNEAKVK DQG- LEGDQG-GENEAKVKLEG- LEGKGGAGQGGYGGLGGQGAGRGAG GENEAKVKLEGNKNEAKVKLEG NEAKVKLEGKG- KNEAKVKLEGNKNEAKVKLEADGGA KG- GKNEAKVKLE- AQGGQGLGGQGESEAKVKLEGNKNE GKNEAKVKLEGNKNEAKVKLEA NEAKVKLEADG- AKVKLEGNGGAGQGGYGGLGSQGAG DG- GESEAKVKLE- RGGYGGQGAGKSEAKVKLEGNKNEA GESEAKVKLEGNKNEAKVKLEG NEAKVKLEGNG- KVKLEGNGGAGQGGYGGLGGQGAGQ NG- GKSEAKVKLEGNKNEAKVK GAGESEAKVKLEGGKNEAKVKLEGN GKSEAKVKLEGNKNEAKVKLEG LEGNG- GGAGQGGYGGLGSQGAGRGAEAEAK NG- GESEAKVKLEGGKNEAKVK VKLEGNKNEAKVKLEGNGGAGQGGY GESEAKVKLEGGKNEAKVKLEG LEGNG- GGLGSQGAGRGGYGGQGAGEAEAKV NG- AEAEAKVKLEGNKNEAKVK KLEGNKNEAKVKLEGNGGAGQGGYG AEAEAKVKLEGNKNEAKVKLEG LEGNG-GEAEAKVKLE- GLGSQGAGRGAEAEAKVKLEGNKNE NG- EAKVKLEGNG- AKVKLEGNGQGGYGGLGGQGAGRGA GEAEAKVKLEGNKNEAKVKLEG AEAEAKVKLE-

[0488]

[0489] GEAEAKVKLEGNNNEAKVKLEGNGG NG- EAKVKLEGN- 78

[0490] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0491] AGQGGYGGLGGQGAGEAEAKVKLEG AEAEAKVKLEGNKNEAKVKLEG GEAEAKVKLE- NNNEAKVKLEGNGGAAQGGQGLGGQ N- EAKVKLEGN- GENEAKVKLEGNNQEAKVKLEGKGG GEAEAKVKLEGNNNEAKVKLEG GEAEAKVKLE- AGQGGYGGLGSQGAGRGGYGGQGAG N- EAKVKLEGNG- ENEAKVKLEGNNQEAKVKLEGEGGA GEAEAKVKLEGNNNEAKVKLEG GENEAKVKLE- GQGGYGGLGSQGAGRGAKNEAKVKL NG- QEAKVKLEGKG- EGKNQEAKVKLEGKGGAGQGGYGGL GENEAKVKLEGNNQEAKVKLEG GENEAKVKLE- GSQGAGRGGYGGQGAENEAKVKLEG KG- QEAKVKLEGEG- ENNEAKVKLEGDQGGYGDLGSQGAG GENEAKVKLEGNNQEAKVKLEG KNEAKVKLE- KGEAKVKLEGKNQEAKVKLEKGKKA EG- QEAKVKLEGK- EAEKKNEKVELK KNEAKVKLEGKNQEAKVKLEGK- AENEAKVKLEGE- AENEAKVKLEGENNEAKVKLEG EAKVKLEG- GKGEAKVKLEGK- EAKVKLEK

[0492] 30 Parallel pA56660 MEIETEGKVEVELEGEAEAKVKLEGK NEAKVKLEGEKGEAKVKLEGD- EIET-KVEVELE- solenoid QGGYGDLGSQGAGNNEAKVKLEGEK NEAKVKLEGKENEAKVKLEGN- AEAKVKLEG-NEAKVKLE- GEAKVKLEGDGGAGQGGYGGLGSQG KNEAKVKLEGDENEAKVKLEGN- KGEAKVKLEGD- AGRGGYGGQGAGKNEAKVKLEGKEN KNEAKVKLEGKKNEAKVKLEGN- NEAKVKLE- EAKVKLEGNGGAGQGGFGGLGGQGA KNEAKVKLEADESEAKVKLEGN- ENEAKVKLEGN- GKNEAKVKLEGDENEAKVKLEGNGG KNEAKVKLEGNKSEAKVKLEGN- KNEAKVKLE- AAQGGQGLGGQGKNEAKVKLEGKKN KNEAKVKLEGNESEAKVKLEGG- ENEAKVKLEGN- EAKVKLEGNGGAGQGGFGGLGGQGA KNEAKVKLEGNEAEAKVKLEGN- KNEAKVKLEGKKNEAKVKL GKNEAKVKLEADESEAKVKLEGNGG KNEAKVKLEGNEAEAKVKLEGN- EGN- AGQGGYGGLGSQGAGRGAKNEAKVK KNEAKVKLEGNEAEAKVKLEGN- KNEAKVKLEADESEAKVKL LEGNKSEAKVKLEGNGGAGQGGYGG KNEAKVKLEGNEAEAKVKLEGN- EGN-KNEAKVKLE- LGSQGAGRGAKNEAKVKLEGNESEAK NNEAKVKLEGNEAEAKVKLEGN- KSEAKVKLEGN- VKLEGGGGAGQGGYGGLGGQGAGKN NEAKVKLEGNENEAKVKLEGN- KNEAKVKLE- EAKVKLEGNEAEAKVKLEGNGQGGY QEAKVKLEGKENEAKVKLEGNG- ESEAKVKLEGG- GGLGGQGAGRGAGKNEAKVKLEGNE QEAKVKLEGEKNEAKVKLEGKG- KNEAKVKLE- AEAKVKLEGNGGAGQGGYGGLGSQG QEAKVKLEGKENEAKVKLEGE- EAEAKVKLEGN- AGRGGYGGQGAKNEAKVKLEGNEAE NEAKVKLEGDKGEAKVKLEGK KNEAKVKLE- AKVKLEGNGGAGQGGYGGLGSQGAG EAEAKVKLEGN- RGGYGGQGAKNEAKVKLEGNEAEAK KNEAKVKLE-

[0493]

[0494] VKLEGNGGAGQGGYGGLGSQGAGRG EAEAKVKLEGN- 79

[0495] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0496] GYGGQGAGNNEAKVKLEGNEAEAKV KNEAKVKLE- KLEGNGGAAQGGQGLGGQGNNEAKV EAEAKVKLEGN- KLEGNENEAKVKLEGNQGGYGDLGS NNEAKVKLEGNEAEAKVKL QGAGNQEAKVKLEGKENEAKVKLEG EGN- NGGAGQGGYGGLGGQGAGRGAGNQE NEAKVKLEGNENEAKVKLE AKVKLEGEKNEAKVKLEGKGQGGYG GN-QEAKVKLE- GLGGQGAGRGAGNQEAKVKLEGKEN ENEAKVKLEGNG- EAKVKLEGEGGAAQGGQGLGGQGNN QEAKVKLEGEKNEAKVKLE EAKVKLEGDKGEAKVKLEGKGGAGQ GKG- GGYGGLGGQGAGNQEAKVKLEKGKK QEAKVKLEGKENEAKVKLE AEAEKKNEKVELK GENE AKVKLEGDKGEAKVKLE GK-QEAKVKLEK-EKVEL

[0497] 31 Parallel pA56662 MIKVEGEGVKVEAEGDAKLEAKAEGE ENKLEAKAEGDKNKLEAKAEGK- IKV-VKVEAE-KLEAKAE- solenoid GNKLEAKAEGKGGAGQGGYGGLGSQ KNKLEAKAEGDKNKLEAKAEGD KLEAKAEGK-QGA- GAGRGAENKLEAKAEGDKNKLEAKA G- ENKLEAKAE-KLEAKAEGK- EGKGGAAQGGQGLGGQGKNKLEAKA GNKLEAKAEGDKNKLEAKAEGK- KNKLEAKAE- EGDKNKLEAKAEGDGGAGQGGYGGL NKLEAKAEGDKNKLEAKAEGD KLEAKAEGDG- GGQGAGQGAGGNKLEAKAEGDKNKL GNKLEAKAE-KLEAKAEGK- EAKAEGKGGAGQGGYGGLGSQGAGR NKLEAKAE-KLEAKAEGD- GGYGGQGAGNKLEAKAEGDKNKLEA AKLEAKAE-KLEAKAE- KAEGDGGAGQGGYGGLGSQGAGRGG LKIEP-KVKRE YGGQGAGGAKLEAKAEGDKNKLEAK AEGSGNLKIEPGKTKKDSKGAKVKRE EKK

[0498] 32 Parallel pA56663 MIKVEGEGVKVEAEGDAKLEAKAEGE KLEAKAEGKENKLEAKAE- IKV-VKVEAE-KLEAKAE- solenoid GGAGQGGYGGLGSQGAGRGGYGGQG KLEAKAEGKKNKLEAKAE- QGAG-KLEAKAEGK- AGGNKLEAKAEGKENKLEAKAEGDG KLEAKAEGDGNKLEAKAE- KLEAKAE-KLEAKAEGK- GAGQGGYGGLGGQGAGQGAGKNKLE KLEAKAEGKGNKLEAKAE- KLEAKAE-KLEAKAE- AKAEGKKNKLEAKAEGDGGAGQGGY KLEAKAEGDGAKLEAKA KLEAKAE-KLEAKAE- GGLGGQGAGKNKLEAKAEGDGNKLE KLEAKAE-KLEAKAE- AKAEGDGGAGQGGYGGLGSQGAGRG KLEAKA-KLEAKAE-

[0499]

[0500] GYGGQGAGKNKLEAKAEGKGNKLEA NLKIEP-AKVKRE

[0501] 80

[0502] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0503] KAEGDGGAGQGGYGGLGGQGAGQG AGKNKLEAKAEGDGAKLEAKAEGDG GAAQGGQGLGGQGKNKLEAKAEGSG NLKIEPGKTKKDSKGAKVKREEKK

[0504] 33 Parallel pA56664 MIKVEGEGVKVEAEGDAKLEAKAEGE KLEAKAEGKENKLEAKAEG- VKVEAEG-KLEAKAEG- solenoid GGAAQGGQGLGGQGGNKLEAKAEGK KNKLEAKAEGKKNKLEAKAEG- KLEAKAEGK-KLEAKAEG- ENKLEAKAEGDGQGGYGGLGGQGAG AGKNKLEAKAEGDGNKLEAKAE KNKLEAKAEGK- RGAGKNKLEAKAEGKKNKLEAKAEG G- KLEAKAEG- DGGAGQGGYGGLGSQGAGRGGYGGQ GAGQGAGKNKLEAKAEGKGNKL AGKNKLEAKAE- GAGKNKLEAKAEGDGNKLEAKAEGD EAKAEG- KLEAKAEG- GGAGQGGYGGLGGQGAGQGAGKNK RGAGKNKLEAKAEGDGAKLEAK GAGQGAGKNKLEAKAE- LEAKAEGKGNKLEAKAEGDGQGGYG AEG KLEAKAEG- GLGGQGAGRGAGKNKLEAKAEGDGA RGAGKNKLEAKAE- KLEAKAEGDGGAGQGGYGGLGGQGA KLEAKAEG- GQGAGKNKLEAKAEGSGNLKIEPGKT AGKNKLEAKAE-NLKIE- KKDSKGAKVKREEKK KVKR

[0505] 34 Parallel pA56665 MIKVEGEGVKVEAEGDAKLEAKAEGE KLEAKAEGKENKLEAKAEGDG- GVKVEAEG-KLEAKAE- solenoid GGAGQGGYGGLGGQGAGRGAGGNKL KLEAKAEGKKNKLEAKAEGDG- KLEAKAEGK- EAKAEGKENKLEAKAEGDGQGGYGG KLEAKAEGDGNKLEAKAE- KLEAKAEGDG- LGGQGAGRGAGKNKLEAKAEGKKNK KLEAKAEGKGNKLEAKA- KLEAKAEGK- LEAKAEGDGGAGQGGYGGLGSQGAG KLEAKAEGDGAKLEAKA KLEAKAEGDG-KLEAKAE- RGGYGGQGAKNKLEAKAEGDGNKLE KLEAKAE-KLEAKAE- AKAEGDQGGYGDLGSQGAGKNKLEA KLEAKA-KLEAKAE- KAEGKGNKLEAKAEGDQGGYGDLGS KLEAKA-KLEAKAE- QGAGKNKLEAKAEGDGAKLEAKAEG NLKIEP-KVKREEK DQGGYGDLGSQGAGKNKLEAKAEGS GNLKIEPGKTKKD SKGAKVKREEKK

[0506] 35 Parallel pA56666 MVKVEGEGVKVKVEGEAKAEAKAEG KIEIKLEGKKNKAEAKAEGD- VKVE-VKVKVE- solenoid EGGAGQGGYGGLGGQGAGGNKIEIKL KIEIKLEGEENKAEAKAEGE- KAEAKAEGE-AGQ-KIEIKLE- EGKKNKAEAKAEGDGGAAQGGQGLG KIEIKLEGDKNKAEAKAEGE- KAEAKAEGD-KIEIKLE- GQGGNKIEIKLEGEENKAEAKAEGEG KIEIKLEGEKNKAEAKAEGE- KAEAKAEGE-KIEIKLE- GAGQGGYGGLGGQGAGRGAGGNKIEI KIEIKLEGDEAKAEAKAEGE KAEAKAEGE-KIEIKLE- KLEGDKNKAEAKAEGEGQGGYGGLG KAEAKAEGE-KIEIKLE- GQGAGRGAGGNKIEIKLEGEKNKAEA AKAEAKAEGE-AGQG-

[0507]

[0508] KAEGEGGAAQGGQGLGGQGGNKIEIK KIEIKLE-GKAKAL-TEV 81

[0509] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0510] LEGDEAKAEAKAEGEGGAGQGGFGG LGGQGAGGNKIEIKLEGSGKAKALPGE TEVDDEGAKVKVEEKK

[0511] 36 Parallel pA56667 MVKVEGEGVKVKVEGEAKAEAKAEG KIEIKLEGKKNKAEAKAEG- VKVE-VKVKVE- solenoid EQGGYGDLGSQGAGGNKIEIKLEGKK KIEIKLEGEENKAEAKAEG- KAEAKAEG-KIEIKLE- NKAEAKAEGDGGAGQGGYGGLGGQG KIEIKLEGDKNKAEAKAEG- KAEAKAEG-KIEIKLE- AGQGAGGNKIEIKLEGEENKAEAKAE KIEIKLEGEKNKAEAKAEG- KAEAKAEG-KIEIKLE- GEGGAGQGGYGGLGGQGAGQGAGG KIEIKLEGDEAKAEAKAEG KAEAKAEG-KIEIKLE- NKIEIKLEGDKNKAEAKAEGEGGAGQ KAEAKAEG-KIEIKLE- GGYGGLGGQGAGGNKIEIKLEGEKNK AKAEAKAEG-KIEIKLE- AEAKAEGEGGAGQGGYGGLGGQGAG GKAKAL-TEVDD RGAGGNKIEIKLEGDEAKAEAKAEGE GQGGYGGLGGQGAGRGAGGNKIEIKL EGSGKAKALPGETEVDDEGAKVKVEE KK

[0512] 37 Parallel pA56668 VKVEGEGVKVKVEGEAKAEAKAEGE GKNKAEAKAEGDGNKIEIKLEGE- VKVE-KVKVE-KAEAKAE- solenoid GNKIEIKLEGKGGAGQGGYGGLGGQG GENKAEAKAEGEGNKIEIKLEGD- KIEIKLEGK-GKNKAEAKAE- AGQGAGKNKAEAKAEGDGNKIEIKLE QGKNKAEAKAEGEGNKIEIKLEG KIEIKLEGE-GENKAEAKAE- GEGGAGQGGYGGLGGQGAGRGAGEN E- KIEIKLEGD- KAEAKAEGEGNKIEIKLEGDGGAAQG AGKNKAEAKAEGEGNKIEIKLEG QGKNKAEAKAE- GQGLGGQGKNKAEAKAEGEGNKIEIK DG KIEIKLEGE- LEGEGGAGQGGYGGLGGQGAGKNKA AGKNKAEAKAE- EAKAEGEGNKIEIKLEGDGGAGQGGY KIEIKLEGDG-EAKAEAKAE- GGLGGQGAGEAKAEAKAEGEGNKIEI KIEIKLE-KAKA-KVKVEEK KLEGSGKAKALPGETEVDDEGAKVKV EEKK

[0513] 38 Parallel pA56669 MVKVEGEGVKVKVEGEAKAEAKAEG KNKAEAKAEGDGNKIEIKLEG- VKVE-KVKVE-KAEAKAE- solenoid EGNKIEIKLEGKGQGGYGGLGGQGAG ENKAEAKAEGEGNKIEIKLEGD- KIEIKLEG-KNKAEAKAE- RGAGKNKAEAKAEGDGNKIEIKLEGE NKAEAKAEGEGNKIEIKLEGEG- KIEIKLEG-ENKAEAKAE- GGAGQGGYGGLGSQGAGRGAENKAE NKAEAKAEGEGNKIEIKLEGDG KIEIKLEGD-NKAEAKAE- AKAEGEGNKIEIKLEGDGGAGQGGYG KIEIKLEGEG-NKAEAKAE- GLGGQGAGQGAGKNKAEAKAEGEGN KIEIKLEGDG-AKAEAKAE- KIEIKLEGEGQGGYGGLGGQGAGRGA KIEIKLEG-KVKVEEK GKNKAEAKAEGEGNKIEIKLEGDGGA

[0514]

[0515] GQGGYGGLGSQGAGRGGYGGQGAGE

[0516] 82

[0517] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0518] AKAEAKAEGEGNKIEIKLEGSGKAKAL PGETEVDDEGAKVKVEEKK

[0519] 39 Parallel pA56673 MMKIKKTENAEVKATGEGGAGQGGY NNEIEVESEKGKNKAELKG- MKIK-EVKATG-NNEIEVE- solenoid GGLGSQGAGRGGYGGQGANNEIEVES KNKAELKGNNNKAELKG- KAELKG-KNKAELK- EKGKNKAELKGEGGAGQGGYGGLGS KNKAELKGNNNKAELKGN- KAELKG-KNKAELK- QGAGRGGYGGQGAKNKAELKGNNNK KNKAELKGNDNKAELKGN- KAELKGN-KNKAELK- AELKGNGGAGQGGYGGLGSQGAGRG KNKAELKGNKNKAELKGN- KAELKGN-KNKAELK- AKNKAELKGNNNKAELKGNGGAGQG KNKAELKGNKNKAELKGN- KAELKGN-KNKAELK- GYGGLGGQGAGKNKAELKGNDNKAE KNKAELKGNKNKAELKGN- KAELKGN-KNKAELK- LKGNGGAGQGGFGGLGGQGAGKNKA KNKAELKGNKNKAELKG- KAELKGN-KNKAELK- ELKGNKNKAELKGNGGAGQGGFGGL ENKAELKGKKNKAELKG- KAELKG-ENKAELK- GGQGAGKNKAELKGNKNKAELKGNG KNKAELKGNKNKAELTGK KAELKG-KNKAELK- QGGYGGLGGQGAGRGAGKNKAELKG NKAELTGK-TTEVTIK- NKNKAELKGNQGGYGDLGSQGAGKN GEAYVKGE KAELKGNKNKAELKGGGGAGQGGYG GLGSQGAGRGGYGGQGAENKAELKG KKNKAELKGEQGGYGDLGSQGAGKN KAELKGNKNKAELTGKGGAGQGGYG GLGSQGAGRGGYGGQGATTEVTIKGG GEAYVKGEGKVEV

[0520] 40 Parallel pA56674 MMKIKKTENAEVKATGEGGAGQGGF NNEIEVESEKGKNKAELKGEG- MKIK-EVKATGE-NNEIEVE- solenoid GGLGGQGAGNNEIEVESEKGKNKAEL KNKAELKGNNNKAELKGNG- KAELKGEG-KNKAELK- KGEGGAGQGGYGGLGSQGAGRGGYG KNKAELKGNNNKAELKG- KAELKGNG-KNKAELK- GQGAGKNKAELKGNNNKAELKGNGG KNKAELKGNDNKAELK- KAELKG-KNKAELK- AGQGGYGGLGSQGAGRGAKNKAELK KNKAELKGNKNKAELKGN- KAELK-KNKAELK- GNNNKAELKGNGGAGQGGYGGLGGQ KNKAELKGNKNKAELKGNG- KAELKGN-KNKAELK- GAGKNKAELKGNDNKAELKGNGGAG KNKAELKGNKNKAELKGNG- KAELKGNG-KNKAELK- QGGYGGLGGQGAGRGAGKNKAELKG KNKAELKGNKNKAELKGG- KAELKGNG-KNKAELK- NKNKAELKGNGQGGYGGLGGQGAGR ENKAELKGKKNKAELKGEG- KAELKGG-ENKAELK- GAGKNKAELKGNKNKAELKGNGGAA KNKAELKGNKNKAELTGKG KAELKGEG-KNKAELK- QGGQGLGGQGKNKAELKGNKNKAEL NKAELTGKG-TTEVTIK- KGNGGAGQGGYGGLGSQGAGRGGYG GEAYVK-EGKVE

[0521] GQGAKNKAELKGNKNKAELKGGGGA

[0522]

[0523] GQGGFGGLGGQGAGENKAELKGKKN

[0524] 83

[0525] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0526] KAELKGEGGAGQGGFGGLGGQGAGK NKAELKGNKNKAELTGKGGAGQGGY GGLGGQGAGTTEVTIKGGGEAYVKGE GKVEV

[0527] 41 Parallel pA56676 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAELKGN- MKIKKTENAEVKA- solenoid KGKNKAELKGEKNKAELKGNGGAGQ NNKAELKGNKNKAELKGN- NNEIEVESEKGK-KNKAELK- GGYGGLGSQGAGRGGYGGQGANNKA NNKAELKGNKNKAELK- KNKAELKGN-NNKAELK- ELKGNKNKAELKGNGGAGQGGYGGL DNKAELKGNKNKAELKGN- KNKAELKGN-NNKAELK- GSQGAGRGGYGGQGAGNNKAELKGN KNKAELKGNKNKAELKGN- KNKAELK-DNKAELK- KNKAELKGNGGAGQGGYGGLGGQGA KNKAELKGNKNKAELK- KNKAELKGN-KNKAELK- GDNKAELKGNKNKAELKGNGGAGQG KNKAELKGNKNKAELKGN- KNKAELKGN-KNKAELK- GYGGLGSQGAGRGGYGGQGAKNKAE KNKAELKGGENKAELKGK- KNKAELK-KNKAELK- LKGNKNKAELKGNGGAAQGGQGLGG KNKAELKGEKNKAELKG KNKAELKGN-KNKAELK- QGKNKAELKGNKNKAELKGNQGGYG ENKAELKGK-KNKAELK- DLGSQGAGKNKAELKGNKNKAELKG KNKAELKG- NGGAGQGGYGGLGSQGAGRGAKNKA KNKAELTGKTTEVTIK- ELKGGENKAELKGKGGAGQGGFGGL GEAYVKGEGKVE GGQGAGKNKAELKGEKNKAELKGNG GAGQGGYGGLGSQGAGRGGYGGQGA GKNKAELTGKTTEVTIKGGGEAYVKG EGKVEV

[0528] 42 Parallel pA56677 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAELKG- MKIKK-AEVKA-NNEIEVES- solenoid KGKNKAELKGEKNKAELKGNGGAGQ NNKAELKGNKNKAELK- KNKAELK-KNKAELKG- GGYGGLGSQGAGRGGYGGQGANNKA NKAELKGNKNKAELK- NNKAELK-KNKAELK- ELKGNKNKAELKGNGGAGQGGYGGL NKAELKGNKNKAELK- NKAELK-KNKAELK- GSQGAGRGGYGGQGAGNNKAELKGN KNKAELKGNKNKAELK- NKAELK-KNKAELK- KNKAELKGNGGAGQGGYGGLGSQGA KNKAELKGNKNKAELK- KNKAELK-KNKAELK- GRGADNKAELKGNKNKAELKGNGGA KNKAELKGNKNKAELK- KNKAELK-KNKAELK- GQGGYGGLGGQGAGRGAGKNKAELK KNKAELKGGENKAELK- KNKAELK-KNKAELK- GNKNKAELKGNGGAGQGGFGGLGGQ KNKAELKGEKNKAELK KNKAELK-ENKAELK- GAGKNKAELKGNKNKAELKGNGGAG KNKAELK-KNKAELK-

[0529]

[0530] QGGYGGLGSQGAGRGGYGGQGAKNK NKAELT-TTEVTIK-AYVK 84

[0531] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0532] AELKGNKNKAELKGNGGAGQGGYGG LGGQGAGKNKAELKGGENKAELKGK GGAGQGGYGGLGSQGAGRGAKNKAE LKGEKNKAELKGNGGAGQGGYGGLG SQGAGRGGYGGQGAGKNKAELTGKT TEVTIKGGGEAYVKGEGKVEV

[0533] 43 Parallel pA56678 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAELKG- MKIKKTENAEVKA- solenoid KGKNKAELKGEKNKAELKGNGGAGQ NNKAELKGNKNKAELKG- NNEIEVESEKGK-KNKAELK- GGYGGLGSQGAGRGANNKAELKGNK NNKAELKGNKNKAELKG- KNKAELKG-NNKAELK- NKAELKGNGGAGQGGYGGLGSQGAG NKAELKGNKNKAELKGN- KAELKG-NNKAELK- RGANNKAELKGNKNKAELKGNQGGY KNKAELKGNKNKAELKGN- KAELKG-NKAELK- GDLGSQGAGDNKAELKGNKNKAELK KNKAELKGNKNKAELKGN- KAELKGN-KNKAELK- GNGGAGQGGYGGLGGQGAGRGAGK KNKAELKGNKNKAELKG- KAELKGN-KNKAELK- NKAELKGNKNKAELKGNGQGGYGGL KNKAELKGGENKAELKG- KAELKGN-KNKAELK- GGQGAGRGAGKNKAELKGNKNKAEL KNKAELKGEKNKAELKG NKAELKG-KNKAELK- KGNGGAGQGGYGGLGSQGAGRGGYG ENKAELKG-KNKAELK- GQGAKNKAELKGNKNKAELKGNGGA KNKAELKG- GQGGYGGLGGQGAGKNKAELKGGEN KNKAELTGKTTEVTIK- KAELKGKQGGYGDLGSQGAGKNKAE GEAYVKGEGKV LKGEKNKAELKGNGGAGQGGYGGLG GQGAGQGAGKNKAELTGKTTEVTIKG GGEAYVKGEGKVEV

[0534] 44 Parallel pA56679 MMKIKKTENAEVKATGENNEIEVESE KNKAELKGEKNKAELKG- MKIKK-AEVKA-NNEIEVES- solenoid KGKNKAELKGEKNKAELKGNGGAGQ NNKAELKGNKNKAELKGNG- KNKAELK-KNKAELKG- GGYGGLGGQGAGRGAGNNKAELKGN NNKAELKGNKNKAELKGNG- NNKAELK-KNKAELKGNG- KNKAELKGNGGAGQGGYGGLGSQGA DNKAELKGNKNKAELKGNG- NNKAELK-KNKAELKGNG- GRGANNKAELKGNKNKAELKGNGGA KNKAELKGNKNKAELKGNG- DNKAELK-KNKAELKGNG- GQGGYGGLGGQGAGDNKAELKGNKN KNKAELKGNKNKAELKGNG- KNKAELK-KNKAELKGNG- KAELKGNGGAGQGGYGGLGGQGAGR KNKAELKGNKNKAELKGNG- KNKAELK-KNKAELKGNG- GAGKNKAELKGNKNKAELKGNGGAG KNKAELKGGENKAELKGKG- KNKAELK-KNKAELKGNG- QGGYGGLGGQGAGRGAGKNKAELKG KNKAELKGEKNKAELKG KNKAELK-ENKAELKGKG- NKNKAELKGNGGAAQGGQGLGGQGK KNKAELK-KNKAELKG- NKAELKGNKNKAELKGNGGAGQGGY AGQ-KNKAELTGKTTEVTIK-

[0535]

[0536] GGLGGQGAGKNKAELKGGENKAELK GEAYVKGEGKVE

[0537] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0538] GKGGAGQGGYGGLGSQGAGRGAKNK AELKGEKNKAELKGNGGAGQGGYGG LGSQGAGRGGYGGQGAGKNKAELTG KTTEVTIKGGGEAYVKGEGKVEV

[0539] 45 Parallel pA56680 MMKIKKTENAEVKATGENNEIEVESE GKNKAELKGEKNKAELKGN- MKIKKTENAEVKA- solenoid KGKNKAELKGEKNKAELKGNGGAGQ ANNKAELKGNKNKAELKGN- NNEIEVESEKG-GKNKAELK- GGYGGLGSQGAGRGANNKAELKGNK GNNKAELKGNKNKAELK- KNKAELKGN-ANNKAELK- NKAELKGNGGAGQGGFGGLGGQGAG GDNKAELKGNKNKAELK- KNKAELKGN-GNNKAELK- NNKAELKGNKNKAELKGNGGAGQGG AGKNKAELKGNKNKAELK- KNKAELK-GDNKAELK- YGGLGGQGAGQGAGDNKAELKGNKN KNKAELKGNKNKAELKGN- KNKAELK-AGKNKAELK- KAELKGNGGAGQGGYGGLGGQGAGK KNKAELKGNKNKAELKGN- KNKAELK-KNKAELK- NKAELKGNKNKAELKGNGGAAQGGQ KNKAELKGGENKAELKGK- KNKAELKGN-KNKAELK- GLGGQGKNKAELKGNKNKAELKGNG KNKAELKGEKNKAELKGN KNKAELKGN-KNKAELK- GAGQGGYGGLGSQGAGRGAKNKAEL ENKAELKGK-KNKAELK- KGNKNKAELKGNQGGYGDLGSQGAG KNKAELKGN-AKNKAELT- KNKAELKGGENKAELKGKGGAGQGG TTEVTIK-GEAYVK-GKVE YGGLGSQGAGRGGYGGQGAKNKAEL KGEKNKAELKGNGGAGQGGYGGLGS QGAGRGGYGGQGAKNKAELTGKTTE VTIKGGGEAYVKGEGKVEV

[0540] 55 Parallel pA56775 MNKIKKTKNSEVKSTGENNEIESKSKK ENKIESTGDNNKIESTG- KIKK-EVKSTG-EIESK- solenoid GKNKIESTGGAGQGGYGGLGSQGAGR KNKIESTGDKNKIESTGQ- KIESTG-ENKIEST-KIESTG- GGYGGQGAGGNENKIESTGDNNKIES ENKIESKGNENKIESTQG- KNKIEST-KIESTGQ- TGGAAQGGQGLGGQGGNKNKIESTGD KNKIESKGDKNKIESTGQ- ENKIESK-KIESTQG- KNKIESTGQGGYGGLGGQGAGRGAG ENKIESKGDKNKIESKG- KNKIESK-KIESTGQ- GNENKIESKGNENKIESTQGGYGDLGS KNKIESKGNKNKIESK- ENKIESK-KIESKG- QGAGGNKNKIESKGDKNKIESTGQGG KNKIESKGNKNKIESK- KNKIESK-KIESK-KNKIESK- YGGLGGQGAGRGAGGNENKIESKGD ENKIESKGNENKIESK KIESK-ENKIESK-KIESK- KNKIESKGGAGQGGYGGLGGQGAGQ KNKIEST-NKIESE-GEIKVS- GAGGNKNKIESKGNKNKIESKGGAGQ GEITSN-VNVTK

[0541] GGYGGLGSQGAGRGAGNKNKIESKGN

[0542]

[0543] KNKIESKGQGGYGGLGGQGAGRGAG

[0544] 86

[0545] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0546] GGENKIESKGNENKIESKGGAGQGGFG GLGGQGAGGNKNKIESTGNKNKIESE GSGEIKVSGSGEITSNGNVNVTKK

[0547] 56 Parallel pA56776 MNKIKKTKNSEVKSTGENNEIESKSKK ENKIESTGDNNKIESTGNG- NKIKK-EVKSTG-NNEIESK- solenoid GKNKIESTGNGQGGYGGLGGQGAGR KNKIESTGDKNKIESTGNG- KIESTGN-ENKIEST- GAGENKIESTGDNNKIESTGNGGAGQ ENKIESKGNENKIESTGNG- KIESTGNG-KNKIEST- GGYGGLGGQGAGRGAGKNKIESTGD KNKIESKGDKNKIESTGNQ- KIESTGNG-ENKIESK- KNKIESTGNGGAGQGGYGGLGGQGA ENKIESKGDKNKIESKGN- KIESTGNG-KNKIESK- GQGAGENKIESKGNENKIESTGNGGA KNKIESKGNKNKIESKGNGQG- KIESTGNQ-ENKIESK- GQGGYGGLGGQGAGRGAGKNKIESK KNKIESKGNKNKIESKGGG- KIESKGN-KNKIESK- GDKNKIESTGNQGGYGDLGSQGAGEN ENKIESKGNENKIESKGN KIESKGNGQG-KNKIESK- KIESKGDKNKIESKGNGGAGQGGYGG KIESKGGG-ENKIESK- LGSQGAGRGGYGGQGAGKNKIESKGN KIESKGN-KNKIEST-NKIESE- KNKIESKGNGQGGYGGLGGQGAGRG SGEIKVS-GEITSN-VNVTK AGKNKIESKGNKNKIESKGGGGAGQG GYGGLGGQGAGENKIESKGNENKIES KGNGGAGQGGYGGLGSQGAGRGGYG GQGAKNKIESTGNKNKIESEGSGEIKV SGSGEITSNGNVNVTKK

[0548] 57 Parallel pA56777 MNKIKKTKNSEVKSTGENNEIESKSKK KNKIESTGNENKIESTGD- NKIKK-EVKST-NEIESKS- solenoid GKNKIESTGNENKIESTGDGGAGQGG NNKIESTGNKNKIESTGD- KNKIEST-ENKIESTGD- YGGLGGQGAGRGAGNNKIESTGNKN KNKIESTGNENKIESKGNG- NNKIEST-KNKIESTGD- KIESTGDGGAGQGGYGGLGGQGAGR ENKIESTGNKNKIESKGD- KNKIESTGNENKIESKGNG- GAGKNKIESTGNENKIESKGNGGAGQ KNKIESTGNENKIESKGDG- ENKIESTGNKNKIESKGD- GGYGGLGSQGAGRGGYGGQGAGENK KNKIESKGNKNKIESKGN- KNKIEST-ENKIESKGDG- IESTGNKNKIESKGDGGAGQGGYGGL KNKIESKGNKNKIESKGN- KNKIESK-KNKIESKGN- GGQGAGQGAGKNKIESTGNENKIESK KNKIESKGGENKIESKGN- KNKIESK-KNKIESKGN- GDGGAAQGGQGLGGQGKNKIESKGN ENKIESKGNKNKIEST KNKIESK-ENKIESKGN- KNKIESKGNGGAGQGGFGGLGGQGA ENKIESKGNKNKIEST- GKNKIESKGNKNKIESKGNGGAGQGG KNKIESEGSGEIKVS- YGGLGSQGAGRGAKNKIESKGGENKI SGEITSN-VNVTK

[0549]

[0550] ESKGNGGAGQGGYGGLGGQGAGRGA

[0551] 87

[0552] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0553] GENKIESKGNKNKIESTGNGGAGQGG YGGLGSQGAGRGGYGGQGAKNKIESE GSGEIKVSGSGEITSNGNVNVTKK

[0554] 58 Parallel pA56778 MNKIKKTKNSEVKSTGENNEIESKSKK KIESTGNENKIEST- NKIKK-EVKST-NEIESK- solenoid GKNKIESTGNENKIESTGGAGQGGYG KIESTGNKNKIEST- KIEST-ENKIEST-KIEST- GLGSQGAGRGAGDNNKIESTGNKNKI KIESTGNENKIESK- KNKIEST-KIEST-ENKIESK- ESTGQGGYGGLGGQGAGRGAGGDKN KIESTGNKNKIESK- KIEST-KNKIESK-KIEST- KIESTGNENKIESKGGAGQGGYGGLGS KIESTGNENKIESK- ENKIESK-KIESK-KNKIESK- QGAGRGGYGGQGAGNENKIESTGNK KIESKGNKNKIESK- KIESK-KNKIESK-KNKIESK- NKIESKGGAGQGGYGGLGGQGAGRG KIESKGNKNKIESK- ENKIESK- AGGDKNKIESTGNENKIESKGGAGQG KNKIESKGGENKIESK- ENKIESKGNKNKIEST- GYGGLGGQGAGGDKNKIESKGNKNKI ENKIESKGNKNKIEST KNKIESEGSGEIKVS- ESKGGAGQGGYGGLGGQGAGRGAGG GEITSN-VNVTK NKNKIESKGNKNKIESKGGAGQGGYG GLGGQGAGRGAGGNKNKIESKGGEN KIESKGGAGQGGYGGLGGQGAGQGA GGNENKIESKGNKNKIESTGGAGQGG YGGLGSQGAGRGGYGGQGAGNKNKI ESEGSGEIKVSGSGEITSNGNVNVTKK

[0555] 59 Parallel pA56779 MNKIKKTKNSEVKSTGENNEIESKSKK ENKIESTGDNNKIEST- KIKK-EVKSTG-EIESKSKK- solenoid GKNKIESTGQGGYGGLGGQGAGRGA KNKIESTGDKNKIEST- KIESTG-ENKIESTGD-KIEST- GGNENKIESTGDNNKIESTGGAGQGG ENKIESKGNENKIEST- KNKIEST-KIEST-ENKIESK- YGGLGSQGAGRGGYGGQGAGNKNKI KNKIESKGDKNKIEST- KIEST-KNKIESK-KIEST- ESTGDKNKIESTGGAGQGGYGGLGSQ ENKIESKGDKNKIESK- ENKIESK-KIESK-KNKIESK- GAGRGGYGGQGAGNENKIESKGNEN KNKIESKGNKNKIESK- KIESK-KNKIESK-KIESKG- KIESTGGAAQGGQGLGGQGGNKNKIE KNKIESKGNKNKIESKG- ENKIESK-KIESKG-KNKIEST- SKGDKNKIESTGGAGQGGYGGLGSQG ENKIESKGNENKIESKG NKIESE-SGEIKVS-GEITSN- AGRGGYGGQGAGNENKIESKGDKNKI VNVTK ESKGGAGQGGYGGLGSQGAGRGGYG GQGAGNKNKIESKGNKNKIESKGQGG

[0556]

[0557] YGGLGGQGAGRGAGGNKNKIESKGN

[0558] 88

[0559] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0560] KNKIESKGGAGQGGYGGLGGQGAGG GENKIESKGNENKIESKGQGGYGGLG GQGAGRGGAGNKNKIESTGNKNKIES

[0561] EGSGEIKVSGSGEITSNGNVNVTKK

[0562] 60 Parallel pA56780 MNKIKKTKNSEVKSTGENNEIESKSKK ENKIESTGDNNKIESTGN- NKIKK-EVKSTG-NNEIESK- solenoid GKNKIESTGNGGAGQGGYGGLGSQGA KNKIESTGDKNKIESTGN- KIESTGN-ENKIEST- GRGAENKIESTGDNNKIESTGNGQGGY ENKIESKGNENKIESTGN- KIESTGN-KNKIEST- GGLGGQGAGRGAGKNKIESTGDKNKI KNKIESKGDKNKIESTGNG- KIESTGN-ENKIESK- ESTGNGGAGQGGYGGLGSQGAGRGG ENKIESKGDKNKIESKGN- KIESTGN-KNKIESK- YGGQGAENKIESKGNENKIESTGNGQ KNKIESKGNKNKIESKGN- KIESTGNG-ENKIESK- GGYGGLGGQGAGRGAGKNKIESKGD KNKIESKGNKNKIESKGG- KIESKGN-KNKIESK- KNKIESTGNGGAGQGGYGGLGSQGAG ENKIESKGNENKIESKGN KIESKGN-KNKIESK- RGGYGGQGAENKIESKGDKNKIESKG KIESKGG-ENKIESK- NGGAGQGGYGGLGSQGAGRGAKNKI KIESKGN-KNKIEST-NKIESE- ESKGNKNKIESKGNGGAGQGGYGGLG SGEIKVS-GEITSN-VNVTK SQGAGRGGYGGQGAKNKIESKGNKN KIESKGGGGAGQGGFGGLGGQGAGEN KIESKGNENKIESKGNGGAGQGGYGG LGGQGAGRGAGKNKIESTGNKNKIESE GSGEIKVSGSGEITSNGNVNVTKK

[0563] 61 Parallel pA56782 MVEVKGEGAEVKVEGEAEAKIELTGE NEAKIELTGEKNEAKIELTG- VEV-AEVKVE-EAKIELTG- solenoid GGAGQGGYGGLGGQGAGQGAGDNE NEAKIELTGKNNEAKIELTG- NEAKIELT-EAKIELTG- AKIELTGEKNEAKIELTGNGGAGQGG NNEAKIELTGDNNEAKIELTG- NEAKIELT-EAKIELTG- YGGLGGQGAGQGAGKNEAKIELTGK KNEAKIELTGKNNEAKIELTG NNEAKIELT-EAKIELTG- NNEAKIELTGNGGAGQGGYGGLGGQ KNEAKIELT-EAKIELTG- GAGQGAGNNEAKIELTGDNNEAKIEL NNEAKIELT-AEAKIELTG- TGNGGAGQGGYGGLGSQGAGRGAKN NEAKIELT-GKAEVKN- EAKIELTGKNNEAKIELTGNGGAGQG KSVDSE-AELK GYGGLGGQGAGNNEAKIELTGDNAEA KIELTGDNNEAKIELTGSGKAEVKNGE

[0564]

[0565] KSVDSEGATAELKEKK

[0566] 89

[0567] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0568] 62 Parallel pA57141 MVWTGEGAEVKVEGEANAVINLTGG EAKIELTGEKNVAVINLT- AVINLT-EAKIELT- solenoid GAGQGGYGGLGSQGTSGRGGLGGQG EAKIELTGKNNVAVINLT- VAVINLT-EAKIELT- AGEDNEAKIELTGEKNVAVINLTGGG EAKIELTGDNNVAVINLT- VAVINLT-EAKIELT- AGQGGYGGLGSQGTSGRGGLGGQGA EAKIELTGKNNVAVINLT VAVINLT-EAKIELT- GNKNEAKIELTGKNNVAVINLTGGGA VAVINLT-EAKIELT- GQGGYGGLGSQGTSGRGGLGGQGAG ANAVINL-EAKIELT- NNNEAKIELTGDNNVAVINLTGGGAG GKATVNN-KKVDSE-TAELK QGGYGGLGSQGTSGRGGLGGQGAGN KNEAKIELTGKNNVAVINLTGGGAGQ GGYGGLGSQGTSGRGGLGGQGAGNN NEAKIELTGDNANAVINLTGDNNEAKI ELTGSGKATVNNGEKKVDSEGATAEL KEKK

[0569] 63 Parallel pA57186 APTITTTGAATVTYTGANLTLTSTSAS TNTATSTGPTSTITLT- ATVTYT-NLTLTS-TSTITLT- solenoid GTSTITLTQGGYGDLGSQGAGGQTNT TNTATSTGATSTITLT- TNTATS-TSTITLT-TNTATS- ATSTGPTSTITLTGGAGQGGYGGLGSQ TNTATSTGNTSTITLT- TSTITLT-TNTATS-TSTITLT- GAGRGAGQTNTATSTGATSTITLTGGA TNKATSTGNNSTITLT- TNKATS-NSTITLT-TNTAKS- GQGGYGGLGSQGAGRGAGDTNTATS TNTAKSTGNNSTITLT- TITLT-TNKAES-TITLT- TGNTSTITLTGGAGQGGFGGLGGQGA TNKAESTGNNSTITLT- TNTAKS-TITLT-TNKAES- GGDTNKATSTGNNSTITLTGGAGQGG TNTAKSTGNNSTITLT- TITLT-TNTATS-TITLT- YGGLGSQGAGRGGYGGQGAGDTNTA TNKAESTGNNSTITLT GTGTA-STITTTS-TTTTTTT KSTGNNSTITLTGGAGQGGYGGLGSQ GAGRGAGDTNKAESTGNNSTITLTGG AAQGGQGLGGQGGDTNTAKSTGNNS TITLTGGAGQGGFGGLGGQGAGGATN KAESTGNNSTITLTGQGGYGGLGGQG AGRGAGGATNTATSTGSNSTITLTGTG TGTAAANSTITTTSGGTTTTTTTA

[0570] 64 Parallel pA57187 TTTTTTTGASTVTVSGANQTLTSTNAT TNTATSTGPTSTITLT- TTTTT-STVTVS-TLTST- solenoid GTSTITLTQGGYGDLGSQGAGGQTNT TNTATSTGTTSTITLT- TSTITLT-TNTATST- ATSTGPTSTITLTGGAGQGGYGGLGSQ TNTATSTGNTSTITLT- TSTITLT-TNTATS-TSTITLT- GAGRGAGQTNTATSTGTTSTITLTGGA TNTATSTGNNSTITLT- TNTATS-TSTITLT-TNTATS- GQGGYGGLGSQGAGRGAGNTNTATS TNTATSTGNNSTITLT- STITLT-TNTATS-TITLT- TGNTSTITLTGGAGQGGFGGLGGQGA TNTAKSTGNNSTIKLT- TNTAKS-TIKLT-TNKAES- GGNTNTATSTGNNSTITLTGGAGQGG TNKAESTGNNSTITLT- TITLT-TNTATS-TITLT- YGGLGSQGAGRGGYGGQGAGDTNTA TNTATSTGNNSTITLT TNTATS-TITLT-GTATA- TSTGNNSTITLTGGAGQGGYGGLGSQ STITTTS-TTTTTTV

[0571] GAGRGAGDTNTAKSTGNNSTIKLTGG

[0572]

[0573] AAQGGQGLGGQGGDTNKAESTGNNS

[0574] 90

[0575] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0576] TITLTGGAGQGGFGGLGGQGAGGATN TATSTGNNSTITLTGQGGYGGLGGQG AGRGAGGATNTATSTGSNSTITLTGTG TATAAANSTITTTSGGTTTTTTVA

[0577] 65 Parallel pA57193 MVTIKQAGIEIEAESDGPIKAEATVGG KINIELEGEKTEANATAT- GIEIEAE-IKAEA-GAG- solenoid AGQGGYGGLGSQGTSGRGGLGGQGA TININLEGEEAKANATAT- KINIELE-KTEANATAT- GQGNKINIELEGEKTEANATATGGGA TININLKGEKAEANATAT- TININLE-EAKANATAT- GQGGYGGLGSQGTSGRGGLGGQGAG TININLEGEEAKANATAI TININLK-KAEANATAT- QGATININLEGEEAKANATATGGGAG TININLE-EAKANATAI-AGQ- QGGYGGLGSQGTSGRGGLGGQGAGQ GAG- GNTININLKGEKAEANATATGGGAGQ TINITLKGEKAEANATA- GGYGGLGSQGTSGRGGLGGQGAGQG TITITLIGNGKATATA-TTELE NTININLEGEEAKANATAIGGGAGQGG YGGLGSQGTSGRGGLGGQGAGQGNTI NITLKGEKAEANATATGAGNTITITLIG NGKATATAGGEAATAGTATTTELELP

[0578] 66 Parallel pA57250 LKLEKTENAEVKVEGENNEVTAKAGK KAKIELKGEGNKAEAK- NAEVKVE-EVTAK- solenoid GKAKIELKGEGNKAEAKGDGGAGQG KIELKGEKNKAEAK- KAKIELK-KAEAK-KIELK- GYGGLGGQGAGNAKIELKGEKNKAE KIELKGEGNKAEAK- KAEAK-KIELK-KAEAK- AKGDGGAGQGGYGGLGSQGAGRGGY KIELKGEKNKAEAK- KIELK-KAEAK-KIELK- GGQGAGNAKIELKGEGNKAEAKGDG KIELKGEGNKAEAK- KAEAK-KIELK-KAEAK- GAGQGGFGGLGGQGAGNAKIELKGEK KIELKGKENKAEAK- KIELK-KAEAK-KIELK- NKAEAKGDGGAGQGGYGGLGGQGA KIELKGKKNKAEAK- KAEAK-KIELK-KAEAK- GRGAGDAKIELKGEGNKAEAKGDGG KIELKGGENKAEAK- KIELK-KAEAK-KIELK- AGQGGFGGLGGQGAGKAKIELKGKEN KIELKGKNNKAEAK- KAEAK-KIELK-KAEAK- KAEAKGDGGAGQGGYGGLGSQGAGR KIELKGNGNKAEAK- KIELK-KAEAK-KIELK- GAKAKIELKGKKNKAEAKGDGGAGQ KIELKGNNNKAEAK- KAEAK-KIELK-KAEAK- GGFGGLGGQGAGKAKIELKGGENKAE KIELKGNNNKAEAK- KIELK-KAEAK-KIELK- AKGDGGAGQGGYGGLGSQGAGRGGY KIELKGNDNKAEAK- KAEA-AKIELK-NKAEAD- GGQGAGKAKIELKGKNNKAEAKGDG KIELKGNKNKAEAK- VKIKVE-ATAKVK GAGQGGFGGLGGQGAGKAKIELKGN KIELKGNKNKAEAK- GNKAEAKGDGGAGQGGYGGLGGQG KIELKGEKNKAEAK-

[0579]

[0580] AGQGAGKAKIELKGNNNKAEAKGDG

[0581] 91

[0582] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0583] GAGQGGYGGLGGQGAGKAKIELKGN KIELKGEKNKAEA- NNKAEAKGDGGAGQGGYGGLGSQGA AKIELKGEKNKAEAD GRGGYGGQGANAKIELKGNDNKAEA KGDGGAGQGGYGGLGSQGAGRGANA KIELKGNKNKAEAKGNGGAGQGGYG GLGGQGAGRGAGNAKIELKGNKNKA EAKGDGGAGQGGYGGLGGQGAGRGA GNAKIELKGEKNKAEAKGNGQGGYG GLGGQGAGRGAGNAKIELKGEKNKAE AKTDDAKIELKGEKNKAEADLKEGQK VKIKVEGGATAKVKPDEKKEIEVEKKI

[0584] K

[0585] 67 Parallel pA57251 LKLEKTENAEVKVEGENNEVTAKAGK KAKIELKGEGNKAEAK- NAEVKVE-EVTAK- solenoid GKAKIELKGEGNKAEAKGDGGAGQG NAKIELKGEKNKAEAK- KAKIELK-KAEAK- GYGGLGGQGAGNAKIELKGEKNKAE AKIELKGEGNKAEAK- NAKIELK-KAEAK-AKIELK- AKGDGGAGQGGFGGLGGQGAGNAKI AKIELKGEKNKAEAK- KAEAK-AKIELK-KAEAK- ELKGEGNKAEAKGDGGAGQGGYGGL DAKIELKGEGNKAEAK- DAKIELK-KAEAK- GGQGAGRGAGNAKIELKGEKNKAEA KAKIELKGKENKAEAK- KAKIELK-ENKAEAK- KGDGGAGQGGYGGLGSQGAGRGGYG KAKIELKGKKNKAEAK- KAKIELKGKKNKAEAK- GQGAGDAKIELKGEGNKAEAKGDGG KAKIELKGGENKAEAK- KAKIELKGGENKAEAK- AGQGGYGGLGSQGAGRGGYGGQGAK KAKIELKGKNNKAEAK- KAKIELKGKNNKAEAK- AKIELKGKENKAEAKGDGGAAQGGQ KAKIELKGNGNKAEAK- KAKIELK-KAEAK- GLGGQGKAKIELKGKKNKAEAKGDQ KAKIELKGNNNKAEAK- KAKIELK-KAEAK- GGYGDLGSQGAGKAKIELKGGENKAE KAKIELKGNNNKAEAK- KAKIELK-KAEAK- AKGDGGAGQGGYGGLGSQGAGRGAK NAKIELKGNDNKAEAK- NAKIELK-KAEAK- AKIELKGKNNKAEAKGDGGAAQGGQ NAKIELKGNKNKAEAK- NAKIELK-KAEAK- GLGGQGKAKIELKGNGNKAEAKGDG NAKIELKGNKNKAEAK- NAKIELK-KAEAK- GAAQGGQGLGGQGKAKIELKGNNNK NAKIELKGEKNKAEAK- NAKIELK-KAEAK- AEAKGDGGAGQGGYGGLGSQGAGRG NAKIELKGEKNKAEAK NAKIELK-KAEAK- AKAKIELKGNNNKAEAKGDGGAGQG DAKIELK-KAEAD- GYGGLGSQGAGRGGYGGQGAGNAKI KVKIKVE-TAKVK-EVEK ELKGNDNKAEAKGDQGGYGDLGSQG AGNAKIELKGNKNKAEAKGNGGAGQ

[0586]

[0587] GGYGGLGSQGAGRGANAKIELKGNK

[0588] 92

[0589] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0590] NKAEAKGDGGAGQGGYGGLGSQGAG RGANAKIELKGEKNKAEAKGNGGAG QGGYGGLGSQGAGRGGYGGQGAGNA KIELKGEKNKAEAKTDDAKIELKGEK NKAEADLKEGQKVKIKVEGGATAKV KPDEKKEIEVEKKIK

[0591] 68 Parallel pA57252 LKLEKTENAEVKVEGENNEVTAKAGK AKIELKGEGNKAEAKG- KLEKT-AEVKVE-EVTAKA- solenoid GKAKIELKGEGNKAEAKGDGGAGQG AKIELKGEKNKAEAKG- AKIELK-KAEAKG-AKIELK- GYGGLGGQGAGNAKIELKGEKNKAE AKIELKGEGNKAEAKG- KAEAKG-AKIELK-KAEAKG- AKGDGGAGQGGYGGLGGQGAGNAKI AKIELKGEKNKAEAKG- AKIELK-KAEAKG- ELKGEGNKAEAKGDGGAGQGGYGGL DAKIELKGEGNKAEAKG- DAKIELK-NKAEAKG- GGQGAGQGAGNAKIELKGEKNKAEA KAKIELKGKENKAEAKGD- KAKIELK-ENKAEAKGD- KGDGGAGQGGYGGLGGQGAGDAKIE KAKIELKGKKNKAEAKGD- KAKIELKGKKNKAEAKGD- LKGEGNKAEAKGDGGAGQGGYGGLG KAKIELKGGENKAEAKG- KAKIELKGGENKAEAKG- GQGAGRGAGKAKIELKGKENKAEAK KAKIELKGKNNKAEAKG- KAKIELKGKNNKAEAKG- GDGGAGQGGYGGLGSQGAGRGAKAK KAKIELKGNGNKAEAKG- KAKIELK-KAEAKG- IELKGKKNKAEAKGDQGGYGDLGSQG KAKIELKGNNNKAEAKG- KAKIELK-KAEAKG- AGKAKIELKGGENKAEAKGDQGGYG KAKIELKGNNNKAEAKG- KAKIELK-KAEAKG- DLGSQGAGKAKIELKGKNNKAEAKGD NAKIELKGNDNKAEAKG- NAKIELK-KAEAKG- GGAGQGGYGGLGSQGAGRGGYGGQG NAKIELKGNKNKAEAKG- NAKIELK-KAEAKG- AKAKIELKGNGNKAEAKGDGGAGQG AKIELKGNKNKAEAKG- AKIELK-KAEAKG-AKIELK- GYGGLGGQGAGQGAGKAKIELKGNN AKIELKGEKNKAEAKG KAEAKG-AKIELK-KAEAK- NKAEAKGDQGGYGDLGSQGAGKAKI AKIELK-NKAEAD-VKIKVE- ELKGNNNKAEAKGDGGAAQGGQGLG ATAKVK GQGNAKIELKGNDNKAEAKGDGGAG QGGYGGLGSQGAGRGANAKIELKGN KNKAEAKGNGGAGQGGYGGLGGQG

[0592]

[0593] AGRGAGNAKIELKGNKNKAEAKGDG

[0594] 93

[0595] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0596] GAGQGGYGGLGGQGAGNAKIELKGE KNKAEAKGNGGAGQGGFGGLGGQGA GNAKIELKGEKNKAEAKTDDAKIELK GEKNKAEADLKEGQKVKIKVEGGATA KVKPDEKKEIEVEKKIK

[0597] 69 Parallel pA57253 LKLEKTENAEVKVEGENNEVTAKAGK KAKIELKGEGNKAEAKG- KLEKT-AEVKVE-EVTAKA- solenoid GKAKIELKGEGNKAEAKGDGGAGQG NAKIELKGEKNKAEAKG- KAKIELK-KAEAKG- GFGGLGGQGAGNAKIELKGEKNKAEA ANAKIELKGEGNKAEAKG- NAKIELK-KAEAKG- KGDGGAGQGGYGGLGSQGAGRGANA GNAKIELKGEKNKAEAKG- ANAKIELK-KAEAKG- KIELKGEGNKAEAKGDGGAGQGGYG GDAKIELKGEGNKAEAKG- GNAKIELK-KAEAKG- GLGSQGAGRGGYGGQGAGNAKIELK GKAKIELKGKENKAEAKG- GDAKIELK-KAEAKG- GEKNKAEAKGDGGAGQGGYGGLGGQ KAKIELKGKKNKAEAKGD- GKAKIELK-ENKAEAKG- GAGDAKIELKGEGNKAEAKGDGGAG KAKIELKGGENKAEAKGD- KAKIELKGKKNKAEAKGD- QGGFGGLGGQGAGKAKIELKGKENKA KAKIELKGKNNKAEAKGDG- KAKIELKGGENKAEAKGD- EAKGDGGAGQGGYGGLGGQGAGQG GKAKIELKGNGNKAEAKGDG- KAKIELKGKNNKAEAKGDG AGKAKIELKGKKNKAEAKGDGGAGQ GKAKIELKGNNNKAEAKGDG- -GKAKIELK-KAEAKGDG- GGYGGLGGQGAGKAKIELKGGENKA GKAKIELKGNNNKAEAKGDG- GKAKIELK-KAEAKGDG- EAKGDGGAGQGGYGGLGGQGAGRGA GNAKIELKGNDNKAEAKGDG- GKAKIELK-KAEAKGDG- GKAKIELKGKNNKAEAKGDGQGGYG GNAKIELKGNKNKAEAKGNG- GNAKIELK-KAEAKGDG- GLGGQGAGRGAGKAKIELKGNGNKA AGNAKIELKGNKNKAEAKGD- GNAKIELK-KAEAKGNG- EAKGDGGAGQGGFGGLGGQGAGKAK QGNAKIELKGEKNKAEAKGN AGNAKIELK-KAEAKGD- IELKGNNNKAEAKGDGGAGQGGYGG QGNAKIELK-KAEAKGN- LGGQGAGKAKIELKGNNNKAEAKGD NAKIELK-KAEAK- GGAGQGGYGGLGGQGAGNAKIELKG DAKIELK-KAEAD- NDNKAEAKGDGGAGQGGYGGLGGQ KVKIKVE-TAKVK GAGRGAGNAKIELKGNKNKAEAKGN GGAGQGGYGGLGGQGAGRGAGNAKI

[0598]

[0599] ELKGNKNKAEAKGDGGAAQGGQGLG

[0600] 94

[0601] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0602] GQGNAKIELKGEKNKAEAKGNGGAG QGGYGGLGSQGAGRGANAKIELKGEK NKAEAKTDDAKIELKGEKNKAEADLK EGQKVKIKVEGGATAKVKPDEKKEIE VEKKIK

[0603] 70 Parallel pA57254 LKLEKTENAEVKVEGENNEVTAKAGK AKIELKGEGNKAEAK- KLEKT-AEVKVE-EVTAKA- solenoid GKAKIELKGEGNKAEAKGDGGAGQG AKIELKGEKNKAEAK- AKIELK-KAEAK-AKIELK- GYGGLGSQGAGRGANAKIELKGEKNK KIELKGEGNKAEAK- KAEAK-KIELK-KAEAK- AEAKGDGGAGQGGFGGLGGQGAGNA KIELKGEKNKAEAK- KIELK-KAEAK-KIELK- KIELKGEGNKAEAKGDGGAGQGGYG KIELKGEGNKAEAK- KAEAK-KIELK-KAEAK- GLGGQGAGQGAGNAKIELKGEKNKA KIELKGKENKAEAK- KIELK-KAEAK-KIELK- EAKGDGGAGQGGYGGLGSQGAGRGG KIELKGKKNKAEAK- KAEAK-KIELK-KAEAK- YGGQGAGDAKIELKGEGNKAEAKGD KIELKGGENKAEAK- KIELK-KAEAK-KIELK- QGGYGDLGSQGAGKAKIELKGKENKA KIELKGKNNKAEAK- KAEAK-KIELK-KAEAK- EAKGDGGAGQGGYGGLGSQGAGRGA KIELKGNGNKAEAK- KIELK-KAEAK-KIELK- KAKIELKGKKNKAEAKGDGGAGQGG KIELKGNNNKAEAK- KAEAK-KIELK-KAEAK- YGGLGSQGAGRGGYGGQGAKAKIEL KIELKGNNNKAEAK- KIELK-KAEAK-KIELK- KGGENKAEAKGDGGAGQGGYGGLGG KIELKGNDNKAEAK- KAEAK-AKIELK-NKAEAD- QGAGKAKIELKGKNNKAEAKGDGGA KIELKGNKNKAEAK- VKIKVE-ATAKVK GQGGYGGLGGQGAGQGAGKAKIELK KIELKGNKNKAEAK- GNGNKAEAKGDGGAGQGGFGGLGGQ KIELKGEKNKAEAK- GAGKAKIELKGNNNKAEAKGDGGAG KIELKGEKNKAEAK QGGYGGLGGQGAGRGAGKAKIELKG NNNKAEAKGDGGAGQGGYGGLGSQG AGRGGYGGQGAGNAKIELKGNDNKA EAKGDGGAGQGGYGGLGSQGAGRGG YGGQGANAKIELKGNKNKAEAKGNG

[0604]

[0605] GAGQGGYGGLGGQGAGNAKIELKGN

[0606] 95

[0607] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0608] KNKAEAKGDGGAAQGGQGLGGQGN AKIELKGEKNKAEAKGNGGAGQGGY GGLGGQGAGNAKIELKGEKNKAEAKT DDAKIELKGEKNKAEADLKEGQKVKI KVEGGATAKVKPDEKKEIEVEKKIK

[0609] 71 Parallel pA57255 LKLEKTENAEVKVEGENNEVTAKAGK KAKIELKGEGNKAEAK- NAEVKVE-EVTAK- solenoid GKAKIELKGEGNKAEAKGDQGGYGD KIELKGEKNKAEAKG- KAKIELK-KAEAK-KIELK- LGSQGAGNAKIELKGEKNKAEAKGDG KIELKGEGNKAEAKG- KAEAKG-KIELK-KAEAKG- GAGQGGYGGLGGQGAGRGAGNAKIE KIELKGEKNKAEAKG- KIELK-KAEAKG-KIELK- LKGEGNKAEAKGDGGAGQGGFGGLG KIELKGEGNKAEAKG- KAEAKG-KIELK-KAEAKG- GQGAGNAKIELKGEKNKAEAKGDGG KIELKGKENKAEAKG- KIELK-KAEAKG-KIELK- AGQGGYGGLGGQGAGQGAGDAKIEL KIELKGKKNKAEAKG- KAEAKG-KIELK-KAEAKG- KGEGNKAEAKGDQGGYGDLGSQGAG KIELKGGENKAEAKG- KIELK-KAEAKG-KIELK- KAKIELKGKENKAEAKGDGGAGQGG KIELKGKNNKAEAKG- KAEAKG-KIELK-KAEAKG- YGGLGSQGAGRGGYGGQGAKAKIEL KIELKGNGNKAEAKG- KIELK-KAEAKG-KIELK- KGKKNKAEAKGDGGAGQGGYGGLG KIELKGNNNKAEAKG- KAEAKG-KIELK-KAEAKG- GQGAGRGAGKAKIELKGGENKAEAK KIELKGNNNKAEAKG- KIELK-KAEAKG-KIELK- GDGGAGQGGYGGLGSQGAGRGGYGG KIELKGNDNKAEAKG- KAEAK-AKIELK-KAEAD- QGAKAKIELKGKNNKAEAKGDGGAG KIELKGNKNKAEAKG- VKIKVE-TAKVK QGGYGGLGSQGAGRGGYGGQGAGKA KIELKGNKNKAEAKG- KIELKGNGNKAEAKGDGGAGQGGYG KIELKGEKNKAEAKG- GLGGQGAGQGAGKAKIELKGNNNKA KIELKGEKNKAEAK EAKGDGGAGQGGFGGLGGQGAGKAK IELKGNNNKAEAKGDGGAGQGGYGG LGSQGAGRGGYGGQGAGNAKIELKG NDNKAEAKGDGGAAQGGQGLGGQG

[0610]

[0611] NAKIELKGNKNKAEAKGNGGAGQGG

[0612] 96

[0613] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0614] YGGLGSQGAGRGGYGGQGAGNAKIE LKGNKNKAEAKGDGGAGQGGYGGLG SQGAGRGANAKIELKGEKNKAEAKGN GGAGQGGFGGLGGQGAGNAKIELKGE KNKAEAKTDDAKIELKGEKNKAEADL KEGQKVKIKVEGGATAKVKPDEKKEI EVEKKIK

[0615] 72 Parallel pA57256 LKLEKTENAEVKVEGENNEVTAKAGK KAKIELKGEGNKAEAKG- NAEVKVE-EVTA-KAKIELK- solenoid GKAKIELKGEGNKAEAKGDGGAGQG AKIELKGEKNKAEAKG- KAEAKG-AKIELK-KAEAKG- GFGGLGGQGAGNAKIELKGEKNKAEA AKIELKGEGNKAEAKG- AKIELK-KAEAKG-AKIELK- KGDGGAGQGGYGGLGSQGAGRGGYG AKIELKGEKNKAEAKG- KAEAKG-AKIELK-KAEAKG- GQGANAKIELKGEGNKAEAKGDQGG AKIELKGEGNKAEAKG- KAKIELK-ENKAEAKG- YGDLGSQGAGNAKIELKGEKNKAEAK KAKIELKGKENKAEAKG- KAKIELKGKKNKAEAKGD- GDGGAGQGGYGGLGGQGAGDAKIEL KAKIELKGKKNKAEAKGD- KAKIELKGGENKAEAKGD- KGEGNKAEAKGDGGAGQGGYGGLGS KAKIELKGGENKAEAKGD- KAKIELKGKNNKAEAKG- QGAGRGGYGGQGAGKAKIELKGKEN KAKIELKGKNNKAEAKG- KAKIELK-KAEAKG- KAEAKGDGGAGQGGFGGLGGQGAGK KAKIELKGNGNKAEAKG- KAKIELK-KAEAKG- AKIELKGKKNKAEAKGDGQGGYGGL KAKIELKGNNNKAEAKG- KAKIELK-KAEAKG- GGQGAGRGAGKAKIELKGGENKAEA KAKIELKGNNNKAEAKG- NAKIELK-KAEAKG- KGDGQGGYGGLGGQGAGRGAGKAKI NAKIELKGNDNKAEAKG- NAKIELK-KAEAKG- ELKGKNNKAEAKGDGGAGQGGYGGL NAKIELKGNKNKAEAKG- NAKIELK-KAEAKG- GGQGAGKAKIELKGNGNKAEAKGDG NAKIELKGNKNKAEAKG- NAKIELK-KAEAKG-YGGL- GAGQGGYGGLGGQGAGRGAGKAKIE NAKIELKGEKNKAEAKG NAKIELK-KAEAK-KIELK- LKGNNNKAEAKGDGGAAQGGQGLGG KAEAD-KIKVE-TAKVK QGKAKIELKGNNNKAEAKGDGQGGY GGLGGQGAGRGAGNAKIELKGNDNK AEAKGDQGGYGDLGSQGAGNAKIELK GNKNKAEAKGNGGAGQGGYGGLGG

[0616]

[0617] QGAGNAKIELKGNKNKAEAKGDGGA

[0618] 97

[0619] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0620] GQGGYGGLGGQGAGQGAGNAKIELK GEKNKAEAKGNGGAGQGGYGGLGGQ GAGRGAGNAKIELKGEKNKAEAKTDD AKIELKGEKNKAEADLKEGQKVKIKV EGGATAKVKPDEKKEIEVEKKIK

[0621] 73 Parallel pA57259 LKLEKTENAEVKVEGENNEVTAKAGK KAEAKGDNAKIELKGE- KLEKT-AEVKVE-EVTAKA- solenoid GKAKIELKGEGGAAQGGQGLGGQGG KNKAEAKGDNAKIELKGE- KAKIELKG-KAEAK- NKAEAKGDNAKIELKGEGGAGQGGY GNKAEAKGDNAKIELKG- NAKIELKGE-KNKAEAK- GGLGSQGAGRGAKNKAEAKGDNAKI KNKAEAKGDDAKIELKG- KIELKGE-GNKAEAK- ELKGEQGGYGDLGSQGAGGNKAEAK GNKAEAKGDKAKIELKG- KIELKG-KNKAEAK- GDNAKIELKGEGGAGQGGYGGLGGQ ENKAEAKGDKAKIELKG- KIELKG-GNKAEAK- GAGQGAGKNKAEAKGDDAKIELKGE KNKAEAKGDKAKIELKG- KIELKG-ENKAEAK-KIELKG- GGAGQGGYGGLGSQGAGRGGYGGQG ENKAEAKGDKAKIELKG- KNKAEAK-KIELKG- AGGNKAEAKGDKAKIELKGKGQGGY NNKAEAKGDKAKIELKG- ENKAEAK-KIELKG- GGLGGQGAGRGAGENKAEAKGDKAK NKAEAKGDKAKIELKGN- NNKAEAK-KIELKG- IELKGKGGAGQGGYGGLGSQGAGRG NNKAEAKGDKAKIELKGN- NKAEAK-KIELKGN- GYGGQGAGKNKAEAKGDKAKIELKG NNKAEAKGDNAKIELK- NNKAEAK-KIELKGN- GGGAGQGGYGGLGGQGAGENKAEAK DNKAEAKGDNAKIELKG- NNKAEAK-KIELK- GDKAKIELKGKGGAGQGGYGGLGGQ KNKAEAKGNNAKIELKG- DNKAEAK-KIELKG- GAGRGAGNNKAEAKGDKAKIELKGN KNKAEAKGDNAKIELKG- KNKAEAK-KIELKG- GGAAQGGQGLGGQGGNKAEAKGDK KNKAEAKGNNAKIELKG- KNKAEAK-KIELKG- AKIELKGNGGAGQGGYGGLGSQGAG KNKAEAKTDDAKIELKG KNKAEAK-KIELKG- RGGYGGQGANNKAEAKGDKAKIELK KNKAEA-AKIELKG- GNQGGYGDLGSQGAGNNKAEAKGDN KNKAEAD-VKIKV-ATAKVK AKIELKGNQGGYGDLGSQGAGDNKAE AKGDNAKIELKGNGGAGQGGYGGLG GQGAGKNKAEAKGNNAKIELKGNGG

[0622]

[0623] AGQGGYGGLGGQGAGKNKAEAKGD

[0624] 98

[0625] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0626] NAKIELKGEGGAGQGGFGGLGGQGAG KNKAEAKGNNAKIELKGEGGAAQGG QGLGGQGKNKAEAKTDDAKIELKGEG GAGQGGYGGLGSQGAGRGGYGGQGA GKNKAEADLKEGQKVKIKVEGGATA KVKPDEKKEIEVEKKIK

[0627] 74 Parallel pA57260 LKLEKTENAEVKVEGENNEVTAKAGK KAEAKGDNAKIELKG- KLEKT-AEVKVE-EVTAKA- solenoid GKAKIELKGEGQGGYGGLGGQGAGR KNKAEAKGDNAKIELKG- KAKIELKG-KAEAK- GAGGNKAEAKGDNAKIELKGEGGAG GNKAEAKGDNAKIELKG- NAKIELKG-KNKAEAK- QGGYGGLGSQGAGRGAKNKAEAKGD KNKAEAKGDDAKIELKG- KIELKG-GNKAEAK- NAKIELKGEGQGGYGGLGGQGAGRG GNKAEAKGDKAKIELKG- KIELKG-KNKAEAK- AGGNKAEAKGDNAKIELKGEGGAGQ ENKAEAKGDKAKIELKGK- KIELKG-GNKAEAK- GGYGGLGSQGAGRGGYGGQGAKNKA KNKAEAKGDKAKIELKGG- KIELKG-ENKAEAK- EAKGDDAKIELKGEGGAGQGGYGGL ENKAEAKGDKAKIELKGK- KIELKGK-KNKAEAK- GGQGAGRGAGGNKAEAKGDKAKIEL NNKAEAKGDKAKIELKGN- KIELKGG-ENKAEAK- KGKGGAGQGGYGGLGGQGAGQGAG GNKAEAKGDKAKIELKGN- KIELKGK-NNKAEAK- ENKAEAKGDKAKIELKGKGGAGQGG GNNKAEAKGDKAKIELKG- KIELKGN-GNKAEAK- YGGLGGQGAGKNKAEAKGDKAKIEL GNNKAEAKGDNAKIELKG- KIELKGN-GNNKAEAK- KGGGQGGYGGLGGQGAGRGAGENKA DNKAEAKGDNAKIELKG- KIELKG-GNNKAEAK- EAKGDKAKIELKGKGGAGQGGYGGL KNKAEAKGNNAKIELKGN- KIELKG-DNKAEAK- GGQGAGRGAGNNKAEAKGDKAKIEL KNKAEAKGDNAKIELKGE- KIELKG-KNKAEAK- KGNGQGGYGGLGGQGAGRGAGGNK KNKAEAKGNNAKIELKG- KIELKGN-KNKAEAK- AEAKGDKAKIELKGNGGAGQGGYGG KNKAEAKTDDAKIELKG KIELKGE-KNKAEAK- LGGQGAGQGAGNNKAEAKGDKAKIE KIELKG-KNKAEA- LKGNGGAGQGGFGGLGGQGAGNNKA AKIELKG-KNKAEAD- EAKGDNAKIELKGNGGAGQGGYGGL VKIKVE-EVEKK

[0628] GGQGAGRGAGDNKAEAKGDNAKIEL

[0629]

[0630] KGNGGAGQGGYGGLGGQGAGKNKA

[0631] 99

[0632] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0633] EAKGNNAKIELKGNGGAGQGGYGGL GGQGAGKNKAEAKGDNAKIELKGEG QGGYGGLGGQGAGRGAGKNKAEAKG NNAKIELKGEGGAGQGGFGGLGGQGA GKNKAEAKTDDAKIELKGEGQGGYG GLGGQGAGRGAGKNKAEADLKEGQK VKIKVEGGATAKVKPDEKKEIEVEKKI

[0634] K

[0635] 75 Parallel pA57474 EVEITGKANVTVKAEGDVDLNINLTGE ENTATAESEGEEVTLNINLTG- ANVTVKA-VDLNINLTG- solenoid GQGGYGGLGGQGAGRGAGENTATAE KNTATAKSKGEENTLNINLTG- YGG-ENTATAES- SEGEEVTLNINLTGEQGGYGDLGSQGA KNTATAESEGEENELNINLTG- EVTLNINLTG-KNTATAKS- GKNTATAKSKGEENTLNINLTGEGGA KNEANAKSKGETSTLNINLKG- ENTLNINLTG-KNTATAES- GQGGFGGLGGQGAGKNTATAESEGEE KGKAKAESESEEQELNINLEG- ELNINLTG-KNEANAKS- NELNINLTGEGGAGQGGYGGLGSQGA EAEAKSKGEEQKLNIKLKG- TSTLNINLKG-KGKAKAES- GRGGYGGQGAGKNEANAKSKGETST KAKAKSEGEEQELKIDLEG- EQELNINLEG-EAEAKS- LNINLKGEGGAGQGGYGGLGGQGAG EAKAKSKGEEQKLTIKLKG- EQKLNIKLKG-KAKAK- RGAGKGKAKAESESEEQELNINLEGEQ KNKAKAESEGEEQELKIDLTG- EQELKIDLEG-EAK- GGYGDLGSQGAGKNEAEAKSKGEEQ ENKAEAKSKGEESELTITLTG- EQKLTIKLKG-KNKAK- KLNIKLKGEGGAGQGGYGGLGSQGAG GKNTAKAESEGEESELKIDLTG- EQELKIDLTG-ENKAE- RGGYGGQGAGKNKAKAKSEGEEQEL QGKNTANAKSKGEESELTITLTG- ELTITLTG-GKNTAKAE- KIDLEGEGGAGQGGYGGLGGQGAGR GDNTANAVSEADESELNINLTG ESELKIDLTG- GAGKNEAKAKSKGEEQKLTIKLKGEQ QGKNTANAKS- GGYGDLGSQGAGKNKAKAESEGEEQE ESELTITLTG-GDNTANAVS- LKIDLTGEGGAGQGGYGGLGGQGAGR ESELNINLTG-NTANAV- GAGENKAEAKSKGEESELTITLTGEGG EGEINVDAD AGQGGFGGLGGQGAGKNTAKAESEG EESELKIDLTGEGGAAQGGQGLGGQG KNTANAKSKGEESELTITLTGAGGAGQ GGFGGLGGQGAGDNTANAVSEADESE

[0636]

[0637] LNINLTGEGGAGQGGYGGLGGQGAG

[0638] 100

[0639] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0640] NNTANAVSEKEGEINVDADAGADVEV EE

[0641] 76 Parallel pA57501 MIKKEGKDIEIEAEGDQEAEAKAEGDG KIKIKLKGEKQKAKAKA- IKKE-IEIEAE-QEAEAKA- solenoid GAGQGGYGGLGSQGAGRGAGNKIKIK TIEIKLKGEEQEAEAEA- KIKIKLK-KQKAKAKA- LKGEKQKAKAKAEGNGGAGQGGFGG KIKIELEGEKQKAKAKA- TIEIKLK-EQEAEAEA- LGGQGAGGNTIEIKLKGEEQEAEAEAE TIEIKLKGEEQEAKAEA- KIKIELE-KQKAKAKA- GNGGAGQGGYGGLGSQGAGRGGYGG KIKIELEGEKQKAEAVAT TIEIKLK-EQEAKAEA- QGAGGNKIKIELEGEKQKAKAKATGN KIKIELE-KQKAEAVAT- QGGYGDLGSQGAGGNTIEIKLKGEEQE NTITINLK-DNEATI- AKAEAEGNQGGYGDLGSQGAGGNKI TTVTVNET KIELEGEKQKAEAVATGNGGAGQGGF GGLGGQGAGGNTITINLKGDGDNEATI GEQKDEGGGTTVTVNETK

[0642] 77 Parallel pA57503 MIKKEGKDIEIEAEGDQEAEAKAEGDG KIKIKLKGEKQKAKAKAE- IKKE-DIEIEAE-QEAEAKAE- solenoid GAGQGGYGGLGGQGAGGNKIKIKLKG TIEIKLKGEEQEAEAEAE- KIKIKLK-KQKAKAKAE- EKQKAKAKAEGNGQGGYGGLGGQGA KIKIELEGEKQKAKAKAT- TIEIKLK-EQEAEAEAE- GRGAGGNTIEIKLKGEEQEAEAEAEGN TIEIKLKGEEQEAKAEAE- KIKIELE-KQKAKAKAT- GGAGQGGYGGLGGQGAGQGAGGNKI KIKIELEGEKQKAEAVAT TIEIKLK-EQEAKAEAE- KIELEGEKQKAKAKATGNGGAGQGG KIKIELE-KQKAEAVAT- YGGLGGQGAGGNTIEIKLKGEEQEAK TITINLK-DNEATI-TVTVNET AEAEGNGGAGQGGFGGLGGQGAGGN KIKIELEGEKQKAEAVATGNGGAAQG GQGLGGQGGNTITINLKGDGDNEATIG

[0643]

[0644] EQKDEGGGTTVTVNETK

[0645] 101

[0646] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0647] 78 Parallel pA57506 IKKEGKDIEIEAEGDQEAEAKAEGDGQ KIKIKLKGEKQKAKAKAE- KKEG-DIEIEAE-QEAEAKAE- solenoid GGYGGLGGQGAGRGAGGNKIKIKLKG TIEIKLKGEEQEAEAEAE- KIKIKLK-KQKAKAKAE- EKQKAKAKAEGNGGAGQGGFGGLGG KIKIELEGEKQKAKAKAT- TIEIKLK-EQEAEAEAE- QGAGGNTIEIKLKGEEQEAEAEAEGNG TIEIKLKGEEQEAKAEAE- KIKIELE-KQKAKAKAT- GAGQGGYGGLGGQGAGQGAGGNKIK KIKIELEGEKQKAEAVAT TIEIKLK-EQEAKAEAE- IELEGEKQKAKAKATGNGGAGQGGY KIKIELE-KQKAEAVAT- GGLGGQGAGRGAGGNTIEIKLKGEEQ GAG-QGA-NTITINLK- EAKAEAEGNQGGYGDLGSQGAGGNKI DNEATI-TTVTVNET KIELEGEKQKAEAVATGNGGAGQGGY GGLGGQGAGGNTITINLKGDGDNEATI GEQKDEGGGTTVTVNETK

[0648] 79 Parallel pA57536 ATTVTDVTDATVTASGSNNSATVTSSS GKAENSGTNNSAELKGE- TTVTDV-ATVTASGS- solenoid GSVSITLSGTNNSAEATGEGGAAQGGQ QTAELSGENNEAELEGE- SATVTS-VSITLSGT- GLGGQGNGKAENSGTNNSAELKGEGG GKAENEGTNNKAKLKGD- SAEATGE-AQGGQG- AGQGGFGGLGGQGAGNQTAELSGEN GEQKAKLKGEKNEAKLEGE- GKAEN-SAELKGE-QTAEL- NEAELEGEGGAGQGGYGGLGSQGAG GEGKAELEGEENKAKLEGE- EAELEGE-GKAEN- RGGYGGQGANGKAENEGTNNKAKLK GEQKAKLKGEKNEAKLEGE- KAKLKGD-GEQKAKLK- GDGGAGQGGYGGLGGQGAGQGAGE GENKAELEGEENKAKLEGE- KNEAKLEGE-GEGKAELE- QKAKLKGEKNEAKLEGEGGAGQGGY ENKAKLEGEENEAELEGEG ENKAKLEGE-GEQKAKLK- GGLGGQGAGQGAGEGKAELEGEENK KNEAKLEGE-GENKAELE- AKLEGEGGAGQGGYGGLGGQGAGRG ENKAKLEGE-ENKAKLE- AGEQKAKLKGEKNEAKLEGEGGAGQ ENEAELEGEG-AENKAELE- GGYGGLGGQGAGRGAGENKAELEGE ENKVKIEG-GDAEIE-EIE ENKAKLEGEGGAGQGGYGGLGSQGA GRGAENKAKLEGEENEAELEGEGGAG QGGYGGLGSQGAGRGGYGGQGAENK

[0649]

[0650] AELEGEENKVKIEGEGDAEIEGEGEIEL

[0651] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0652]

[0186] In embodiments, BSRPs of the beta solenoid kind disclosed herein comprise two sheets that, together, encompass 70% or more of all residues as part of a beta strand, wherein the beta strand is identified by DSSP4 secondary structure analysis of the model. In embodiments, said sheets are in contact with each other, forming the folded core of the solenoid.

[0653]

[0187] In embodiments, BSRPs of the beta solenoid kind disclosed herein comprise one sheet that encompasses 70% or more of all residues as part of a beta strand, wherein the beta strand is identified by DSSP4 secondary structure analysis of the model. In embodiments, said sheet is folded upon itself, forming two planes (FIGs.53A-D). In embodiments, the strands that bridge the two planes of said sheet have negative bend residues at the turning point between the two planes (FIG. 53B) As used herein, a “negative bend residue” refers to an amino acid residue characterized by DSSP4 as being bent (“S” under the “ dssp struct summary.bend” column in the DSSP4 mmCIF-formatted output) and having with negative chirality

[0654]

[0655] under the “ dssp struct summary. chirality” column in the DSSP4 mmCIF-formatted output).

[0656]

[0188] In embodiments, BSRPs of the beta solenoid kind disclosed herein comprise at least one solenoid spine of 4 or more residues. As used herein, a “solenoid spine” refers to a network of residues that can be found in a given beta solenoid model such that it has the following characteristics (FIGs. 53B and 53C):

[0657] i. The network edges are pairwise alpha carbon distances of 5.8 A or less

[0658] ii. The network has at least 3 residues

[0659] iii. All residues are negative bend residues

[0660] iv. None of the residues are the first one, second one or third one in the sequence v. If N is the length of the sequence, none of the residues are in the N-4, N-3, N-2, N- 1 or N sequence position

[0661] vi. For each residue, whose position in the sequence is i, the C alpha carbon of residue i-3 and the C alpha carbon of i+5 is between 7.2 A and 10 A.

[0662] vii. For each residue, whose position in the sequence is i, the angle between the vector from the C alpha carbon of said spine residue, to the C alpha carbon of the i-3

[0663] 103

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[0665] residue and the vector from the C alpha carbon of said spine residue, to the C alpha carbon of the i+5 residue, is between 30° and 60°.

[0666] viii. None of the residues are consecutive in sequence

[0667] ix. 60% of all residues within a -7 and +8 residue window of spine residues are in beta strand conformation (by DSSP4 secondary structure analysis of the structure model).

[0668]

[0189] In embodiments, BSRPs of the beta solenoid kind disclosed herein comprise a structural N-terminal structural cap and a C-terminal structural cap (FIG. 53D). As used herein, given a solenoid that has one or more solenoid spines or glycine solenoid spines, a “structural N-terminal cap” refers to a sequence segment that precedes the first solenoid spine residue in the protein sequence, as long as this segment also has the following characteristics:

[0669] i. the segment is 4 or more amino acids in length

[0670] ii. the segment is at most 85% identical to the segment of the same length preceding the second solenoid spine residue in the protein sequence that belongs to the same spine.

[0671]

[0190] As used herein, given a solenoid that has one or more solenoid spines or glycine solenoid spines, a “structural C-terminal cap” refers to a sequence segment that is after the last solenoid spine residue in the protein sequence, as long as this segment also has the following characteristics:

[0672] i. the segment is 4 or more amino acids in length

[0673] ii. the segment is at most 85% identical to the segment of the same length that is after the second to last solenoid spine residue in the protein sequence that belongs to the same spine.

[0674]

[0191] Without being bound by theory, it is thought that the structural arrangement described above, with one sheet folded over, or two closely-packed sheets, as well as a solenoid spine, emulates the local structure of a beta sheet crystalline phase, and therefore any sequence predicted to form said structure will also have a high chance of being able to form beta sheet crystalline phase once processed.

[0675] 104

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[0677]

[0192] Without being bound by theory, it is thought that the presence of N- and C-terminal structural caps may prevent or reduce the end-to-end aggregation of BSRPs of the beta solenoid kind after expression and before processing.

[0678]

[0193] In embodiments, the BSRP is of the antiparallel beta solenoid class, comprising two sheets that, together, encompass 70% or more of all residues as part of a beta strand, wherein the beta strand is identified by DSSP4 secondary structure analysis of the model, and said sheets are composed of antiparallel hairpins (FIGs. 54A and 54B). As used herein, an antiparallel hairpin refers to a polypeptide segment in a mode composed of the following DSSP4 secondary structure elements: strand, followed by a loop, followed by a strand, where said strands are paired to each other, in other words, the elements of strand followed by a loop, and followed by a strands are on the same plane.

[0679] III. Linkers

[0680]

[0194] In embodiments, the BSRPs disclosed herein comprise a linker having an amino acid sequence having at least 80% identity to any one of SEQ ID NOs: 122-158, 175-194, 1143, and 1775-2063. In embodiments, the BSRPs disclosed herein comprise a linker having an amino acid sequence having at least 80% identity to any one of SEQ ID NOs: 122-125, 128-139, 146 and 1775-2063.

[0681]

[0195] As illustrated in FIG. 1A, linkers connect beta strands of the beta strand reservoir polypeptides disclosed herein. Without being bound by theory, it is thought that by separating individual beta strands, linkers allow the flexibility for the beta strands to fold into tertiary structures. Illustrative tertiary structure types include: (i) a structure with dihedral pseudo symmetry, (ii) a beta solenoid structure comprising beta solenoid motifs, and (iii) an anti-parallel beta sheet structure. When the beta strand reservoir polypeptides transition into a beta sheet crystalline phase, as shown in FIG. ID, the presence of linkers, in particular linkers at least 6 amino acids in length and possessing predominantly non-strand and non-helix secondary structure as determined by in silico analyses, impart an elastic character to the beta sheet crystalline phase.

[0682]

[0196] The disclosure provides beta strand reservoir polypeptides disclosed herein which comprise an alternating arrangement of beta strands and linkers. In embodiments, the beta strand

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[0685] reservoir polypeptides disclosed herein comprise “n” number of beta strands and “n-1” number of linkers, wherein each of the linkers is placed between two of the “n” beta strands.

[0686]

[0197] In embodiments, the beta strand reservoir polypeptides that are determined to exhibit a tertiary structure with a dihedral symmetry comprise four or more beta domains and a linker between each of the beta domains. As described above, in embodiments of the beta strand reservoir polypeptides disclosed herein that are determined to exhibit a tertiary structure with internal dihedral pseudo-symmetry, one or more domains comprises a configuration of Lx-(S-L)y-Sz, wherein S is a beta strand and L is a linker, wherein x is 0 or 1, y is any one of 4 through 12, and z is 0 or 1. In embodiments, L is a loop.

[0687]

[0198] In embodiments, the beta strand reservoir polypeptides that are determined to exhibit a tertiary beta solenoid structure comprise at least four beta strands and a linker between each of the beta strands. In embodiments, the linker is a loop.

[0688]

[0199] In embodiments, the linker comprises a region of a naturally occurring protein, such as, a silk protein. In embodiments, the linker comprises a region of a silk fibroin protein. In embodiments, the linker comprises a region of a spider silk protein, such as, a spidroin. In embodiments, the linker comprises a region of a major ampullate spidroin protein. In embodiments, the major ampullate spidroin protein is a major ampullate spidroin (MaSp) 1 or MaSp 2.

[0689]

[0200] In embodiments, the linker comprises about 7 amino acid residues to about 75 amino acid residues, for instance, about 7 amino acid residues, about 10 amino acid residues, about 15 amino acid residues, about 20 amino acid residues, about 25 amino acid residues, about 30 amino acid residues, about 35 amino acid residues, about 40 amino acid residues, about 45 amino acid residues, about 50 amino acid residues, about 55 amino acid residues, about 60 amino acid residues, about 65 amino acid residues, about 70 amino acid residues, or 75 amino acid residues, inclusive of all values and subranges therebetween.

[0690]

[0201] In embodiments, the linker comprises at least 20% glycine (G) residues, for instance about 25% glycine residues, about 30% glycine residues, about 35% glycine residues, about 40% glycine residues, about 45% glycine residues, about 50% glycine residues, about 55% glycine residues, at least 60% glycine residues, at least 65% glycine residues, at least 70% glycine residues, at least 106

[0691] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0692] 75% glycine residues, about 80% glycine residues, at least 85% glycine residues, at least 90% glycine residues, at least 95% glycine residues, or 100% glycine residues. In embodiments, the linker comprises at least 50% glycine (G) residues. In embodiments, the linker does not comprise one or more of the following: (i) an amino acid sequence of (GA)nX, wherein X is any amino acid, and n>5, (ii) an amino acid sequence of An, wherein n >5, and (iii) an amino acid sequence of (AG)n, wherein n >5.

[0693] Compositions and Methods of Preparation thereof

[0694]

[0202] The disclosure provides compositions, comprising any one of the non-naturally occurring, beta strand reservoir polypeptides disclosed herein. The disclosure further provides polynucleotides encoding any one of the beta strand reservoir polypeptides disclosed herein. In embodiments, the polynucleotide is at least 50% identical (for instance, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100%, inclusive of all values and subranges that lie therebetween) to a polynucleotide of any one of SEQ ID NOs: 1061-1142 and 2848. The disclosure further provides expression cassettes comprising any one of the polynucleotides disclosed herein.

[0695]

[0203] The disclosure provides host cells comprising any one of the beta strand reservoir polypeptides disclosed herein, any one of the polynucleotides disclosed herein, or any one of the expression cassettes disclosed herein. The type of host cell is not limited and may be any cell or cell line suitable for protein expression. In embodiments, the host cell is a bacterial cell, such as, for example, E. coli. In embodiments, the host cell is Bacillus sublilis. Corynebacterium glutamicum, Pseudomonas fluorescens. or a species of Streptomyces. In embodiments, the host cell is a eukaryotic cell or cell line. Eukaryotic cell lines include mammalian cell lines, such as human and animal cell lines, insect, plant, or fungal cell lines. Non-limiting examples of such cells or cell lines generated from such cells include Be HROC277, COS, CHO (e.g., CHO-S, CHO-K1, CHO-DG44, CHO-DUXB11, CHO-DUKX, CHOK1SV), VERO, MDCK, WI38, V79, B14AF28-G3, BHK, HaK, NSO, 5P2 / 0-Agl4, HeLa, HEK293 (e.g., HEK293-F, HEK293-H, HEK293-T), and perC6 cells as well as insect cells such as Spodoptera fugiperda (Sf, e.g., Sf9), or fungal cells such as Saccharomyces. Pichia, Aspergillus, Trichoderma and Schizosaccharomyces.

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[0698]

[0204] The disclosure provides methods of preparing any one of the beta strand reservoir polypeptides disclosed herein, the method comprising expressing the polypeptide in a host cell. The disclosure provides a powder, e.g., a lyophilized powder, comprising any one of the beta strand reservoir polypeptides disclosed herein, or any one of the polynucleotides disclosed herein. Without being bound by theory, it is thought that the beta strand reservoir polypeptides disclosed herein are amenable to being lyophilized and resuspended thereafter. The beta strand reservoir polypeptides disclosed herein may therefore be provided in a powder, e.g., a lyophilized powder that can be resuspended into solution and further processed.

[0699]

[0205] In embodiments, the powder comprising any one of the beta strand reservoir polypeptides disclosed herein, or any one of the polynucleotides disclosed herein is generated by drying processes other than lyophilization, such as, for example, via the use of a convection oven.

[0700]

[0206] In embodiments, the disclosure provides a liquid, semi-solid, gel-like or slurry composition or a composition comprising inclusion bodies, comprising any one of the beta strand reservoir polypeptides disclosed herein, or any one of the polynucleotides disclosed herein.

[0701]

[0207] In embodiments, the disclosure provides methods of preparing any one of the beta strand reservoir polypeptides disclosed herein, the method comprising expressing the polypeptide in a host cell, such that the polypeptide is present in one or more inclusion bodies. In embodiments, the method comprises, (i) providing a host cell culture, wherein the host cells express any one of the beta strand reservoir polypeptides disclosed herein, and wherein the BSRPs are present in inclusion bodies in the host cell, and (ii) centrifuging the cell culture to obtain a cell pellet comprising inclusion bodies. The cell pellet may be directly dissolved in formic acid to advantageously facilitate purification of the BSRP and formation of the fiber (e.g. by spinning) at the same time.

[0702]

[0208] In embodiments, the disclosure provides methods of preparing any one of the beta strand reservoir polypeptides disclosed herein, the method comprising expressing the polypeptide in a host cell, such that the polypeptide is present in the cytoplasm of the host cell. In embodiments, the method comprises, (i) providing a host cell culture, wherein the host cells express any one of the beta strand reservoir polypeptides disclosed herein and wherein the BSRPs are present in the

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[0705] cytoplasm of the host cell, (ii) lysing the cells, and (iii) centrifuging the cell culture to obtain a supernatant comprising the expressed BSRP.

[0706]

[0209] The disclosure provides methods of preparing any one of the beta strand reservoir polypeptides disclosed herein, the method comprising expressing the polypeptide using a cell-free expression system.

[0707]

[0210] Without being bound by theory, it is thought that when resuspended into formic acid, beta strand reservoir polypeptides described herein unfold, thus putting them in a state where they are ready to be drawn or spun into a fiber. The relative ease with which the beta strand reservoir polypeptides are able to be resuspended in environmentally friendly solvents, like formic acid and the lack of gelling or other premature intermolecular association of the polypeptides within the (e.g. formic acid) dope prior to spinning are unexpected and superior properties of the beta strand reservoir polypeptides disclosed herein.

[0708] Beta Sheet Crystalline Phase

[0709]

[0211] The disclosure provides, for instance, a beta sheet crystalline phase produced from a plurality of the non-naturally occurring, beta strand reservoir polypeptides disclosed herein. In embodiments, the phase comprises a 3-dimensional stack of a plurality of 2-dimensional beta sheets, as shown in FIGs. 2A-2B. In embodiments, the beta sheet crystalline phase disclosed herein comprises a 3-dimensional stack of at least 2 (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10) 2-dimensional beta sheets.

[0710]

[0212] The presence of intermolecular interactions between beta sheets may be detected using infrared spectroscopy. For instance, the presence of intermolecular interactions between beta sheets, which may indicate the presence of stacked beta sheets in a beta sheet crystalline phase, may be assessed using infrared spectroscopy. In embodiments, the presence of an amide I band between 1622 cm'1and 1627 cm'1indicates the presence of intermolecular interactions between beta sheets. In embodiments, the amide I band is centered at or around 1624 cm'1. Further details are provided in Hu et al.. Macromolecules, Vol. 39, No. 18, 2006, the contents of which are incorporated herein by reference in its entirety.

[0711]

[0213] The 2-dimensional beta sheets disclosed herein are formed by a plurality of beta strands. In embodiments, at least 2 beta strands of each beta sheet are derived from separate beta strand 109

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[0713] reservoir polypeptides. Without being bound by theory, it is thought that upon processing via the methods disclosed herein, the tertiary structure of the beta strand reservoir polypeptide is denatured. In embodiments, the denaturation results in the extension of the beta strand reservoir polypeptide (see red line in FIG. 2A) such that there is unfolding of the tertiary structure as assessed by X-ray diffraction or other suitable methods. Upon drawing and / or spinning, the extension of the beta strand reservoir polypeptides facilitates the formation of 2-dimensional beta sheets comprising one or more beta strands from different polypeptides. In embodiments, one or more beta strands in a 2-dimensional beta sheet belongs to a different polypeptide chain. In embodiments, every beta strand in a 2-dimensional beta sheet belongs to a different polypeptide chain.

[0714]

[0214] In embodiments, the plurality of beta strands of the beta sheet point in the same direction, forming a parallel beta sheet. In embodiments, the plurality of beta strands of the beta sheet point in opposite directions in an alternating manner, forming an anti-parallel beta sheet. In embodiments, the plurality of beta strands of the beta sheet comprises a mixture of strands that point in the same direction and strands that point in the opposite directions. In embodiments, the beta sheet crystalline phase comprises 2-dimensional parallel beta sheets. In embodiments, the beta sheet crystalline phase comprises 2-dimensional anti-parallel beta sheets. In embodiments, the beta sheet crystalline phase comprises 2-dimensional beta sheets with a mixture of parallel and anti-parallel strand pairings. In embodiments, the beta sheet crystalline phase comprises a mixture of 2-dimensional parallel beta sheets, 2-dimensional anti-parallel beta sheets, and / or 2-dimensional beta sheets with a mixture of parallel and anti-parallel strand pairings. In embodiments, one or more beta strands that form a beta sheet have the same sequence. In embodiments, one or more beta strands that form a beta sheet have different sequences. Without being bound by theory, it is thought that the presence of linkers, in particular linkers at least 6 amino acids in length and possessing predominantly non-strand and non-helix secondary structure as determined by in silico analyses, in the beta strand reservoir polypeptides disclosed herein imparts an elastic character to the multi-chain assembly.

[0715]

[0215] The disclosure provides any article or composition that comprises the beta sheet crystalline phase disclosed herein.

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[0718] Articles Comprising a Beta Sheet Crystalline Phase

[0719]

[0216] The present disclosure provides, for example, compositions and articles comprising the beta sheet crystalline phase described herein. In embodiments, a beta sheet crystalline phase described herein comprises a polypeptide having a sequence according to any one of SEQ ID NOs 1-84, or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity thereto. In embodiments, a beta sheet crystalline phase described herein comprises a polypeptide having a linker sequence according to any one of SEQ ID NOs 122-125, 128-139, 146, or 1775-2063, or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity thereto. Exemplary compositions and articles include fabrics (e.g., consumer fabrics and performance fabrics), fibers, filters and filtration devices, medical devices, packaging and inert materials used with pharmaceuticals and biopharmaceutical products, sealants, and vents.

[0720]

[0217] In embodiments, a composition or article described herein is substantially free of (e.g., is free of) silk or silk analogs. In embodiments, a composition or article described herein comprises less than 1%, 0.1%, or 0.01% silk or silk analogs. In embodiments, a composition or article described herein is substantially free of (e.g., is free of) naturally occurring silk protein. In embodiments, a composition or article described herein is substantially free of (e.g., is free of) polypeptides having at least about 30% (for instance, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, or about 70%) identity to a naturally occurring silk protein. In embodiments, a composition or article described herein is substantially free of (e.g., is free of) polypeptides having at least about 70% identity to a naturally occurring silk protein. In embodiments, a composition or article described herein is substantially free of (e.g., is free of) polypeptides having at least about 80% identity to a naturally occurring silk protein. In embodiments, a composition or article described herein is substantially free of (e.g., is free of) polypeptides having at least about 90% identity to a naturally occurring silk protein.

[0721]

[0218] In embodiments, a composition or article described herein comprises a fiber. In embodiments, the length of the fiber is greater than 10 times the width of the fiber. The fiber may comprise a beta sheet crystalline phase described herein. The fiber may have a circular crosssection, an oval cross-section, a square cross-section, a rectangular cross-section, or any desired cross-sectional shape. The cross section of a fiber may be described by its width and thickness. For example, in a perfectly circular cross section, the width and thickness have the same value,

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[0724] and in an oval cross section, the width is greater than the thickness. In embodiments, the fiber has a diameter of 10-50 um, 50-500 um, or 500-1000 um. In embodiments, the fiber has a width of 600-3000 um and / or a thickness of 100-200 um. In embodiments, a composition or article described herein comprises a nanofiber. In embodiments, the nanofiber has a diameter of 1-100 nm, 1-10 nm, or 10-100 nm.

[0725]

[0219] In embodiments, a composition or article described herein comprises a fibrous article. The fibrous article comprises fibers. The fibrous article may further comprise a non-fibrous material. In embodiments, the fibrous article has a fibrillar organization (e.g., mesoscale microfibrillar organization) in its solid state. In embodiments, the fibrous article is a formic acid-cast film.

[0726]

[0220] In embodiments, a composition or article described herein comprises a filament which is a continuous fiber. In embodiments, the filament has a length that is generally measured in meters and can be tens, hundreds or even thousands of meters long. To render the filament into a staple fiber, the filament may be cut or sheared, for example, using a guillotine or rotatory blade mechanism such as available from the DM & E Company located in Shelby, N. C. The desired cut length is determined by which staple process e.g., long or short staple process) and the type of spinning process to be used. In embodiments, when blending different types of staple fiber together to form a spun yarn, the length of the staple fiber are similar, for example the average staple lengths can be within 10% or less of each other. In other embodiments, when blending different types of staple fiber together to form a spun yam, the length of the staple fiber can be greater than 10% of each other.

[0727]

[0221] In embodiments, a composition or article described herein comprises a yarn which comprises a plurality of filament fibers combined into a linear member.

[0728] I. Consumer fabrics and performance fabrics

[0729]

[0222] In embodiments, the composition or article comprising a beta sheet crystalline phase is a consumer fabric or a performance fabric. For instance, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use in fabric or textile apparel for consumer fabrics or performance fabrics applications having suitable durability, breathability, hand, thermal performance, projectile penetration resistance and elasticity for applications such as protective outerwear, garments, footwear, gloves, and accessories (see for example, e.g.,

[0730] 112

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[0732] US9513088B2, US444351 l, US8607476, and US9944044B2, US11105039B2, US10286234B2, US10286234B2, US11105039B2, US7816289B2, and US8769722B2).

[0733]

[0223] Articles, for example, textiles, can be produced from a yam described herein using any known method for producing textiles from yarns. The textile can be produced via known knitting, weaving or nonwoven methods to produce knit textiles, woven textiles, nonwoven textiles or fleece textiles. Suitable examples of textile making methods can include, for example, warp knitting, weft knitting, circular knitting, flatbed knitting, seamless knitting, broad loom, narrow width weaving, belting, rapier weaving, shuttle weaving, air jet weaving, water jet weaving, projectile loom weaving, and jacquard weaving. Suitable examples of nonwoven methods can include, for example, needle punching, hydroentanglement, wet-laid and meltblown. Textiles can be produced using one or more yarns of the present disclosure or the textiles can comprise the disclosed yams and one or more other yams.

[0734]

[0224] In embodiments, the composition or article (e.g., protective garment) comprising a beta sheet crystalline phase is a laminate of a nanofiber layer or a microporous membrane attached to a woven or knit textile of a garment. The beta sheet crystalline phase fibers may be part of the woven or knit textile itself where the elements of the textile may be the fibers themselves, or polyfilament yams comprised in whole or in part by the fibers. Alternatively, the nanofiber layer or membranes may comprise beta sheet crystalline phase fibers, and may be attached to textiles that do not comprise the beta sheet crystalline phase. Further still the beta sheet crystalline phase fibers-based textile may be attached to a membrane or nanofiber layer which does not comprise beta sheet crystalline phase fibers. Yet further still the means of attachment may include an adhesive which in turn may also be comprised of the beta sheet crystalline phase. Thus, in embodiments, one or more of the nanofiber layer or microporous membrane, the textile, or the means of attachment e.g., an adhesive) may be comprised in whole or in part of a beta sheet crystalline phase, a fiber, a nanofiber web, a film, or an article of manufacture described herein.

[0735] II. Cables and Cable Assemblies

[0736]

[0225] In embodiments, the composition or article comprising a beta sheet crystalline phase is a cable or cable assembly. For instance, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use in the reinforcement, load bearing, or electromagnetic 113

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[0738] interference isolation in cables, tubular assemblies and / or cable assemblies. The cables may be suitable for high-speed transmission of data. The cable assembly may comprise a cable and a reinforcing layer around the cable. In embodiments, the fibers may form a braided mechanical reinforcement. In embodiments, the web or film may be applied via a spiral tape wrap as a mechanical reinforcement, e.g., for use in signal transduction cables, test cables for aerospace, semiconductor, land system or precision testing, musical instruments or land system applications, or medical applications, (see for example, e.g., US7700872B2, US11501893B2, and US10240994B1). In embodiments, the cable or cable assembly comprising the fiber, the nanofiber web, the film, or the article of manufacture described herein may include other dispersed elements such as metal ligands, particles, or other inorganics. The other dispersed elements may change the dielectric or electromagnetic character of the fiber, the nanofiber web, the film, or the article of manufacture described herein such that they can provide suitable characteristics of signal shielding or enhanced mechanical performance.

[0739] III. Electronic Components and Seals

[0740]

[0226] In embodiments, the composition or article comprising a beta sheet crystalline phase is an electronic component or a seal. For instance, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use in electronic component applications having thermal insulative, chemical resistance, or dielectric properties suitable for use in electrochemical devices such as fuel cells, sensors, membrane electrodes, thermal insulation for electronics, or electronics (see for example, e.g., US11251453B2, US11380927B2, US7491453B2, US7998624B2, US20240258613A1, and US20230087216A1).

[0741]

[0227] As an example, a wet (liquid phase) ionomer mixture or composite wet mixture may be deposited on a side of a nanofiber web described herein to render an interior volume. The deposition of the wet ionomer mixture or composite wet mixture forms a wet ionomer layer or composite layer with the web or within and on top of the web. The ionomer may be a proton or anion-conducting polymer (e.g., an unreinforced ionomer mixture). In embodiments, the ionomer substantially impregnates the nanofiber web to render an interior volume of the of the microporous membrane layer substantially occlusive (see for example, e.g., U. S. Pat. No. RE 37,307) thereby forming the composite wet layer. The wet ionomer layer or composite wet layer may be conveyed

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[0744] via a roll feed and / or roll winder to a dryer and substantially dried to form a fiber reinforced composite film. The composites may include a protective outer top coat layer which may be another polymer such as silicone or may be a film described herein, which provides for protection and transport.

[0745]

[0228] In embodiments, the composites may find particular use in fuel cells, gas exchange membranes, and biochemical power sources for environmental or biomedical applications. In these cases, the membrane and electrode assembly may be positioned between an anode and a cathode, separating two media to enable the cell's function. In this configuration, the cell can operate such that the flow of ionic species across the semipermeable membrane generates an electrical current in a circuit connecting the anode and cathode. This current can be utilized for power generation, detection or sensing purposes, as in microbial electrochemical fuel cells or implanted glucose sensors or electrochemical fuel cells. Alternatively, an external current can be applied to drive ions across the membrane to initiate or sustain a reaction, as seen in electrochemical cells that find use in flow batteries or energy storage devices. Such devices may be configured as suited to enable function in the art with additional optionally to include, but not limited to, spacers which may often be permeable, porous, microporous, and / or fibrous or webs. These may also include redox polymers and / or enzymes. In embodiments, the redox polymer and / or more than one enzyme can be used in each electrolysis layer. In embodiments, this layer may be covered by a non-fouling coating. This coating or film layer may be in the form of a film described herein or a composite thereof.

[0746]

[0229] In embodiments, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be created with other mineral or silicate fillers including those that take the form of an aerogel. The thus created blend compositions may then be formed into low density matrices with said filler particles such that they form a conformable low dusting, low thermal, or electrical conductivity insulating sheet for use in a variety of applications in the field of electronics. Further said sheets can be combined with a conductive layer (e.g., a graphite or a copper film) such that they have value in dissipating localized heat or electrical charge spatial as may valuable in a variety of electronics applications.

[0747] IV Fibers and Constructs thereof

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[0750]

[0230] In embodiments, the composition or article comprising a beta sheet crystalline phase is a fiber or a construct comprising a fiber. For instance, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use with the required tensile properties, liquid permeability, and conformability in a fiber for industrial applications where fibers and / or high performance fibers and constructs thereof find typical application such as ropes, architectural fabrics, composite reinforcements, sports equipment, or musical instrument strings, or seals (see, for example, e.g., US7296394B2, US7501356B, US6737158B1, US7217876B2, and US12025226B2).

[0751]

[0231] The fiber may be a monofilament fiber and multifilament fiber. A plurality of fibers may be combined to form a “bundle”. When different types of fibers are combined to form a bundle, it may be referred to herein as a “composite bundle.” A plurality of bundles may be combined to form a “bundle group”. A plurality of bundle groups may be combined to form a “rope”.

[0752]

[0232] In embodiments, the bundles may be wrapped by the protein nanofiber web or membrane, or coated by the protein coating.

[0753]

[0233] In embodiments, a composition described herein (e.g., a nanofiber web) is subjected to a compression step. In embodiments, the compression step is used to alter the porosity of the composition. In further embodiments, the fiber bundles, or woven constructions thereof or the nanofiber web or membrane may be selectively compressed and disposed into a composite with a secondary material. In embodiments, the secondary material may be an elastomer such as a natural or synthetic rubber, a thermoset polymer, and / or a thermoplastic polymer. The secondary material may be combined with the fiber bundles, or woven constructions thereof or the nanofiber web or membrane by any known methods in the art such as lamination, compression molding, vacuum impregnation, liquid coating or prepreg.

[0754] V. Filters and Filtration Devices

[0755]

[0234] In embodiments, the composition or article comprising a beta sheet crystalline phase is a filter or a filter device. For example, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use in filtration applications having mesh or sieving openings of macro or micro scale dimensions with the mechanical strength, conformability, stability in fluids, and robustness suitable to fabricate devices such as filter bags, filter cartridges,

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[0758] or flat filter discs or panels, or pleated filter panels which find use, for example, in removing particles and other contaminants from a variety of fluids including liquids, gasses, or other fluid streams therefrom (see for example, e.g., US10350529B2, US10994237B2, US9480953B2, and US9205359B2). In embodiments, a fiber described herein may be knit or woven to form the mesh or sieving opening of the filter or filter device.

[0759]

[0235] In embodiments, a filter media sheet composite comprises a support layer and a layer comprising the beta sheet crystalline phase. In embodiments, a filter sheet media composite comprises a nanofiber web described herein in a composite with a woven or non-woven support. In embodiments, a filtration device comprises a cartridge that comprises the beta sheet crystalline phase. The cartridge may further comprise suitable plastic or metal materials.

[0760]

[0236] Yet another filter embodiment may be a filter bag. In some emobdiments, a filter bag comprises a porous membrane adjacent to a filter media.

[0761] VI. Medical Devices

[0762]

[0237] In embodiments, the composition or article comprising a beta sheet crystalline phase is a medical device. For example, a fiber, a nanofiber web, a film, or an article of manufacture may be configured for use in medical device applications such as use in the form of a suture, deployment aid, coating, or tube of membrane woven or nonwoven fiber for use in a stent or graft having suitable mechanical characteristics, biocompatibility, biodegradation fiber size and pore or opening sizes to provide the desired clinical benefit (see for example, e.g., US9545300B2, US9808605B2, US11147698B2, and US 10111741B2). In embodiments, the beta sheet crystalline phase may be bioabsorbable or inert.

[0763]

[0238] In embodiments, the medical device may include a beta sheet crystalline phase in the form of a fiber, a nanofiber web, a film, or an article of manufacture combined with other materials in a multi-layer composite. The multilayer composite may include a fluoropolymer, such as an expanded polytetrafluoroethylene (ePTFE) or a polyester, a silicone, a urethane, another biocompatible polymer, DACRON® (polyester), bioadsorbable systems, copolymers, or combinations and subcombinations thereof. In embodiments, the beta sheet crystalline phase may be in the form of a coating on a hydrophobic substrate or a hydrophilic substrate, such as a fluoropolymer or glass. In embodiments, the water contact angle of the coated substrate may be 117

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[0765] at or above 90°. In embodiments, the water contact angle of the coated substrate may be at or below 90°.

[0766]

[0239] As an example, a stent may comprise a stent frame that is attached to a covering material comprising the beta sheet crystalline phase in the form of a fiber or plurality of fibers constructed into a knit or woven textile, a non-woven mat or collection of fibers in a web or nanofiber web, or in the form of a film, or in an article thereof which may include a multilayer composite. In embodiments, the covering material may be manufactured using techniques such as, but not limited to, extrusion, expansion, heat-treating, sintering, knitting, weaving, chemically treating, laminating, spinning and the like.

[0767] VII. Combination medical devices that may have therapeutic effect

[0768]

[0240] In embodiments, the composition or article comprising a beta sheet crystalline phase is a combination medical device that may have a therapeutic effect. For example, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use in a combination medical device that may have anti-thrombogenic, anticoagulatory, or accelerated healing therapeutic effects having suitable porosity, compound composition, and placement in a combination medical device to have effect (see, e.g., US9320890B2, EP2680894B1, or US9415193B2, US10736999B2, or US11529441B2, US20220233299A1, US20220008049A1, and US20210315682A1).

[0769]

[0241] An embodiment can be a film material permeable to a therapeutic compound, or a fiber, nanofiber web or membrane that may be impregnated or coated with the beta sheet crystalline phase, into which may be admixed a therapeutic compound (e.g., a small molecule or macromolecule). Said resulting coated film, fiber or membrane may become less permeable and / or impermeable to the therapeutic compound. In some instances, the permeability of the article may not change.

[0770]

[0242] In an example, the beta sheet crystalline phase may be modified with covalently attached heparin or impregnated with one or more therapeutic compounds that may be released in situ to promote wound healing or reduce tissue inflammation. In one or more embodiments, the therapeutic compound may be a corticosteroid, a human growth factor, an anti-mitotic agent, an antithrombotic agent, or dexamethasone sodium phosphate.

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[0773]

[0243] In other embodiments, the therapeutic compounds may include small molecule drugs, large molecule drugs, medicaments, cardiovascular agents, chemotherapeutics, antimicrobials, antibiotics, anesthetics, hemostatics, antihistamines, antitumors, antilipids, antifungals, antimycotics, antipyretics, vasodilators, hypertensive agents, oxygen free radical scavengers, vitamins, antivirals, analgesics, antiproliferatives, anti-inflammatories, diagnostic agents, visualization agents, angiographic contrast agents, phase contrast agents, and radiopaque agents, or thrombolytics intended to facilitate the breakup of thrombus, anticoagulants such as heparin, intended to prevent thrombosis and combinations thereof. The therapeutic composition may be an anti-inflammatory steroid such as dexamethasone sodium phosphate, dexamethasone acetate, dexamethasone, and / or beclomethasone dipropionate.

[0774]

[0244] Yet other therapeutic compounds include, but are not limited to, antirestenotic drugs including, but not limited, to pimecrolimus, cytochalasin, dicumarol, cyclosporine, latrunculin A, methotrexate, tacrolimus, halofuginone, mycophenolic acid, genistein, batimistat, dexamethasone, cudraflavone, simvastatin, prednisolone, doxorubicin, bromopyruvic acid, carvedilol, mitoxantrone, tranilast, etoposide, hirudin, trapidil, mitomycin C, abciximab, cilostazol, irinotecan, estradiol, diaziquone, dipyridamole, melatonin, colchicine, nifedipine, vitamin E, paclitaxol, diltiazem, vinblastine, verapamil, vincristine, rapamycin, angiopeptin, everolimus, heat shock proteins, zotarolimus, nitroglycerin, and prednisone.

[0775]

[0245] An embodiment can be a material impermeable to a therapeutic compound placed on at least one surface of the therapeutic-containing, coated film material as a “capping layer” to prevent movement of the therapeutic compound through or out of the impermeable material. The material for the “capping layer” can be formed of a polymer such as a silicone, a material containing the beta sheet crystalline phase, or another material. Depending on the embodiment, the capping layer material may be applied either to a portion of the coated film material or all of the film material. The portion of the coated film material which may be not covered by the capping layer material preferentially elutes the therapeutic compound when exposed to fluids. The capping layer material may be applied over the coated film material after the film material may be applied to a substrate.

[0776]

[0246] In embodiments of a combination medical device or apparatus, the device may comprise the beta sheet crystalline phase in some form in combination with woven / non woven and porous

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[0779] membrane constructs may be configured such that the device is used to encapsulate directly or indirectly with cells, for instance therapeutic cells of human or other origin to accomplish its function.

[0780] VIII. Packaging and / or inert materials used with pharmaceutical and biopharmaceutical products and in their production

[0781]

[0247] In embodiments, the composition or article comprising a beta sheet crystalline phase is a packaging and / or inert material used with pharmaceutical and biopharmaceutical products, and in their production. For instance, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use as an inert material used with pharmaceutical and biopharmaceutical products wherein such materials can serve, e.g., to reinforce elastomers, provide films or reinforced films for sealing and or provide particulate ingress barriers that maintain sterility (see, e.g., US10478563B2, US6451396B1, US6673455B2 EP1680332B1, US20030211264A1, US9078858B2, US7213336B2, US6793746B2, US8435019B2, US10526367B2, US20240042348A1, WO2024163661A1, and US11224853B2).

[0782]

[0248] For example, a membrane media of a cap for a vial may include a membrane composite wherein the layer or layers may in whole or part be comprised by a fiber, nanofiber web or membranes, woven, or composite thereof, as described herein.

[0783]

[0249] In further articles used with pharmaceutical and biopharmaceutical products the protein nanofiber web or membrane, fibrous article, film or coating described herein may be used as a contact surface on the interior or exterior of a tube, seal or plunger. It may also serve as a matrix or support for a filler particle such as a mesoporous sorbent which may be functionalized to bind or unbind proteins or impurities as in affinity chromatography as used to manufacture, alter or purify drug products as for example in downstream processing of cellular slurries in the production of biopharmaceuticals.

[0784] IX. Vents and devices used in the field and application of venting of containers and enclosures

[0785]

[0250] In embodiments, the composition or article comprising a beta sheet crystalline phase is a vent. For example, a fiber, a nanofiber web, a film, or an article of manufacture described herein may be configured for use in devices for venting such that the devices provide the suitable 120

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[0787] characteristics and configuration to allow for pressure equalization and or sound transmission while inhibiting ingress of liquid or particulate contamination (see, e.g., US8858681B2, US9038773B2, US20200329289A1, US8011539B2, and US9,622,331 B2).

[0788]

[0251] For example, an acoustic cover assembly may comprise a cover material and an acoustic gasket. The cover material and / or the acoustic gasket may comprise the beta sheet crystalline phase described herein.

[0789]

[0252] In embodiments, membranes or films may be porous or non-porous so as to provide for gas exchange and pressure relief so as to have suitable configuration for use in venting packaged volatile liquids, or to provide ventilation to gear boxes as seen across a variety of automotive and industrial applications, or to provide water proofing to electronic enclosures in base stations or mobile electronics.

[0790]

[0253] In embodiments, the films, webs, or coatings may be deployed on their own or after coating with an oleophobic or hydrophobic polymer coating suitable for the application they are slated for.

[0791] X. Cosmetic Compositions

[0792]

[0254] In embodiments, the composition or article comprising a beta sheet crystalline phase is a cosmetic composition. In embodiments, the cosmetic compound may include whitening agents, hair dyes or pigments, anti-aging peptides, skin conditioners, vitamins, fragrances, UV-protective compounds, and antioxidants. The cosmetic composition may be in a cosmetically acceptable medium for the enhancement of keratin-containing materials, mucosa, or teeth. In embodiments, the cosmetic composition includes the beta crystalline phase and at least one cosmetically acceptable auxiliary or excipient. Additionally or alternatively, the cosmetic composition may include conditioning agents, moisturizers, surfactants, colorants, anti-aging compounds, whitening agents, and vitamins. In embodiments, the beta sheet crystalline phase is capable of stabilizing or delivering the cosmetic compound to a keratin surface.

[0793]

[0255] In embodiments, the cosmetic compound is produced by mixing a BSRP with one or more cosmetically active compounds or effect substances; inducing self-assembly or aggregation of the BSRP to form stabilized structures or particles containing the one or more cosmetically active compounds; and combining the resulting mixture or particles with a cosmetically acceptable carrier or auxiliary to form the cosmetic composition. Further, a protein-rich phase may be phase 121

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[0795] separated from a protein-poor phase to produce microbeads or nanoparticles, followed by incorporating those beads or nanoparticles into a cosmetically acceptable base. In embodiments, the microbeads or nanoparticles entrap or encapsulate a hydrophobic or hydrophilic cosmetically active compound, providing controlled or sustained release upon application (e.g. to hair, skin, mucosa, or teeth). Further examples of forms and commercial fields of use are shown and described on the W. L. Gore website at gore.com / or goremedical.com / .

[0796] XL Microporous Membranes

[0797]

[0256] In embodiments, microporous membranes comprising a beta sheet repeat polymer may be produced by any suitable phase separation or pore-forming technique, including, without limitation: non-solvent induced phase separation (NIPS), vapor induced phase separation (VIPS), evaporation induced phase separation (EIPS), temperature induced phase separation (TIPS), combinations thereof, and related casting / coagulation processes. For example, in VIPS, a cast film may be exposed to a controlled humidity atmosphere (optionally followed by immersion in a nonsolvent bath) to induce phase separation. In EIPS, partial evaporation of solvent from a cast film may increase polymer concentration prior to or during phase separation. In TIPS, the polymer may be dissolved in a solvent at elevated temperature and phase separation may be induced by controlled cooling, optionally in the presence of a diluent or non-solvent.

[0798]

[0257] In embodiments, the polymer may be dissolved or dispersed in one or more solvents selected from acids, polar protic solvents, polar aprotic solvents, halogenated solvents, ionic liquids, and mixtures thereof, provided the solvent system is capable of forming a castable composition. Non-limiting examples of solvents include formic acid, acetic acid, trifluoroacetic acid, hexafluoroisopropanol (HFIP), dimethyl sulfoxide (DMSO), N, N-dimethylformamide (DMF), N, N-dimethylacetamide (DMAc), N-methyl-2-pyrrolidone (NMP), and mixtures thereof. In various embodiments, the non-solvent (coagulant) may be water, alcohols (e.g., methanol, ethanol, isopropanol), ketones, esters, or other liquids that are miscible with the solvent and that induce phase separation of the beta sheet repeat polymer, including mixtures of non-solvents. The composition and temperature of the coagulation bath, the residence time in the bath, and the use of multi-stage coagulation baths (e.g., sequential baths of differing composition) may be selected to tailor pore size, porosity, and membrane asymmetry.

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[0801]

[0258] In embodiments, the castable composition may further comprise one or more additives to promote or control pore formation, membrane morphology, or mechanical properties. Such additives can include, for example, pore formers / porogens (e.g., polyethylene glycol, polyvinylpyrrolidone, salts, sugars, soluble particulates), salts, non-solvent additives, viscosity modifiers, surfactants, nanoparticles, fibers, or plasticizers. In embodiments, pores may additionally or alternatively be introduced by leaching of a soluble additive after membrane formation, by gas foaming, by particulate templating, by stretching / expansion, or by combinations of these techniques.

[0802]

[0259] Process variables that may be adjusted to tailor the membrane include, without limitation: polymer concentration, solvent composition, additive content, casting thickness, casting temperature, evaporation time prior to coagulation, humidity during VIPS, coagulation bath composition and temperature, quench rate (e.g., for TIPS), post-coagulation washing conditions, drying conditions (air drying, vacuum drying, freeze-drying, supercritical drying), and posttreatments (e.g., annealing, solvent exchange, crosslinking, crystallization-promoting treatments, surface modification, coating, or lamination). In embodiments, the membrane may be formed as a free-standing film, supported on a substrate, or integrated into a composite (e.g., laminated to a textile, nonwoven, mesh, or porous support layer) to provide breathable barrier structures, filtration media, separators, medical barrier materials, and other articles.

[0803] Methods of Making Articles Comprising a Beta Sheet Crystalline Phase

[0804]

[0260] Articles comprising the beta sheet crystalline phase disclosed herein may be made by any suitable method.

[0805]

[0261] Suitable methods of making fiber include methods wherein a fiber is produced from a solution or suspension in liquid individually as a single filament or continuously as a beam including wet spinning, dry spinning, air gap wet spinning, or pultrusion, such as the process used by insects. Furthermore, fibers or yams can also be spun from solutions to form webs comprised of a plurality of random or partially aligned fibers in spinning processes such as electrospinning, blow spinning, or jet spinning. Further, fibers and polyfilament constructs can be formed from the solid state via processes of lubricated extrusion or thermal extrusion in processes such as capillary 123

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[0807] fiber or yam extrusion, meltspinning, spun bonding, meltblowing, or via roll compaction or wet laying of shear fibrillated materials. In embodiments, fiber may be produced by extrusion. In embodiments, the fiber is produced by extrusion through the syringe needle, producing a beta sheet crystalline phase.

[0808]

[0262] For example, a nanofiber web may be made as follows. A droplet comprising beta strand reservoir polypeptides may be positioned at the tip of a syringe needle. An electric potential may be applied to the droplet, thereby producing electrospun nanofibers. The electrospun nanofibers may be collected, e.g., on an aluminum sheet positioned a suitable distance from the syringe tip. In embodiments, a constant volume flow rate of (e.g., about 0.2 ml / h) may be maintained, e.g., using a syringe pump. The electric potential may be adjusted so that a stable jet is obtained through the Taylor cone.

[0809]

[0263] Wet spinning of fibers may be performed as follows. A solution comprising beta strand reservoir polypeptides may be produced, for example by stirring the proteins into formic acid (e.g., to a concentration of about 40wt% of protein in acid). A syringe may be used to draw up the solution. The syringe (e.g., a 1 mL syringe) may comprise a dispensing tip of 22-25 gauge (e.g., 22 gauge) and / or a length of about 0.5-1.5 inch. The syringe may be placed onto a syringe pump. The syringe pump may be placed above a coagulation bath, so that the syringe tip has an air gap between 0.5 and 5mm, e.g., 1-3 mm. The coagulation bath may be chosen from, e.g., methanol, water, or a mixture of alcohol and water. The solution may be expelled from the syringe, e.g., at a rate of 1.4 pL / hr to 3.2 mL / min, e.g., 0.5-3.2 mL / min, thereby producing spun fibers in the coagulation bath. The fibers may be removed from the coagulation bath and allowed to dry, e.g., in a laboratory hood after hanging with tension.

[0810]

[0264] Wet-spun fibers may be stretched. For instance, a sample may be hand stretched or drawn, e.g., to about 2x or about 3x its original length. The drawn fiber (or an unstretched fiber) may be dried in a vacuum oven, e.g., at about 180 mbar, at about 150°C, for about 30 minutes. The same drying process may also be used for an unstretched fiber.

[0811]

[0265] In embodiments, a beta sheet crystalline phase described herein may be readily recycled. For instance, the beta sheet crystalline phase may be subjected to denaturation in a solvent, and then restructured. In embodiments, the beta sheet crystalline phase is subjected to a first round of 124

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[0813] denaturing and restructuring, a second round of denaturing and restructuring, and optionally a third or more rounds of denaturing and restructuring. In embodiments, a beta sheet crystalline phase described herein does not experience a loss of mechanical properties during a denaturation and restructuring procedure, or experiences a loss of mechanical properties that is superior to a silk undergoing the same process.

[0814] Methods of Testing Fibers Comprising a Beta Sheet Crystalline Phase

[0815]

[0266] The present disclosure provides, for instance, methods for assessing fiber formation for sub-milligram quantities of protein or polymer in the presence of volatile and / or corrosive organic solvents. The disclosure also provides, for instance, methods for automation of testing to enable screening large numbers or protein sample candidates.

[0816]

[0267] In a droplet drawing test, a protein solution may be evaluated for fiber forming or spinnability as follows. A solution of protein in a volatile organic solvent may be provided, e.g., by dissolving the protein in the solvent. A droplet (e.g., of 5 ul or less) of the solution may be placed on a flat surface, e.g., a fluorinated film surface. The droplet may be in a convective air stream, e.g., allowing the solution concentration to increase over time. A rod may be brought into contact with the surface of the droplet. The rod may be withdrawn from the droplet. Fiber formation between the rod and the droplet may be observed, e.g., optically or via a force transducer. This process may be repeated, e.g., for about 15 minutes or until the liquid has evaporated or an observed fiber is formed. The presence or absence of a fiber may be noted.

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[0819] EXAMPLES

[0820]

[0268] The following examples, which are included herein for illustration purposes only, are not intended to be limiting.

[0821] Example 1: De Novo Design and Engineering of BSRPs Determined to Exhibit a Tertiary Structure Having Internal Dihedral Pseudo-Symmetry

[0822]

[0269] Beta domains were designed using ProteinGenerator, which can be directed to create protein backbones with specific secondary structure content (for further details, see Lisanza, etal., “Multistate and functional protein design using RoseTTAFold sequence space diffusion”, Nat Biotechnol 1-11 (2024), the contents of which are incorporated by reference in its entirety). ProteinGenerator was set up to produce protein homodimers with a high percentage of beta-strand content. Thousands of homodimers were generated and filtered using various filter metrics available within a customized software package built on top of the Rosetta macromolecular modeling suite (for further details, see Leman, et al. “Macromolecular modeling and design in Rosetta: recent methods and frameworks”, Nat Methods 17, 665-680 (2020), the contents of which are incorporated by reference in its entirety), ^diffusion was used to create C2 symmetric dimers, which were used as input for RPXdock to obtain dihedral multimers (for further details, see Watson, et al. “De novo design of protein structure and function with ^diffusion''. Nature 620, 1089-1100 (2023) and Sheffler, et al. “Fast and versatile sequence-independent protein docking for nanomaterials design using RPXDock”, PLOS Computational Biology 19, el010680 (2023), the contents of which are incorporated by reference in their entirety).

[0823]

[0270] Sequences for the dihedral multimer backbones were generated using ProteinMPNN (for further details, see Dauparas, et al. “Robust deep learning-based protein sequence design using ProteinMPNN”, Science eadd2187 (2022), the contents of which are incorporated by reference in its entirety). ProteinMPNN takes as input the 3D structure of only the backbone, and outputs a user-specified number of sequences of equal length which the model believes have high likelihood of folding into the input structure. In addition to the backbone, input was provided to ProteinMPNN specifying various parameters, including “temperature”, which controls how sequences are sampled.

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[0826]

[0271] Resulting sequences were validated in silico using Colabfold for structure prediction using default parameters, with the following changes: A) the sequence prediction was carried out with the input sequence only, B) no template was searched for or used, C) only the top three of the five prediction models was used, D) only the top model as ranked by average pLDDT was minimized. Once a Colabfold-based model is produced for each beta domain sequence, the per-position metric pLDDT was used to rank designs. Designs with high average pLDDT (>70) were manually inspected to ensure that the high-confidence structure model had the intended dihedral symmetry (e.g. D4 or De).

[0827]

[0272] Next, the beta domains in a multimer model (e.g. eight or twelve beta domains in the case of D4 or De multimers, respectively) were linked using a polypeptide linker selected from a prespecified set of linker sequences, for example, any one or more of SEQ ID NOs: 122-125, 128-139 and 146, to obtain a single polypeptide sequence, that is: a BSRP with a tertiary structure having internal dihedral pseudo-symmetry. In some cases, further in silico validation was carried out for the resulting BSRPs as discussed above. BSRPs that showed high pLDDT (average value >70) for beta domains were selected for cellular expression. In some instances, the resulting BSRPs were again used as input for ProteinMPNN as discussed above. In some instances, the linkers of the BSRPs were changed while retaining the same beta domains to generate new BSRPs.

[0828]

[0273] The amino acid sequences of the BSRPs determined to exhibit a tertiary structure having internal dihedral pseudo-symmetry are set forth in Table 1. The DSSP (Define Secondary Structure of Proteins) program was used to identify the beta strands within each of these BSRPs.

[0829] Example 2: De Novo Design and Engineering of BSRPs Determined to Exhibit a Tertiary Beta Solenoid Structure

[0830]

[0274] RF diffusion was used to generate BSRPs with beta solenoid tertiary structures.

[0831]

[0275] Capping domains were generated using RF diffusion or a combination of RF diffusion and ProteinGenerator to add residues to the N- and C-termini of the beta solenoid structures described above. The capping domains were designed to prevent the ends of the BSRP from forming intermolecular beta strand-beta strand interactions.

[0832]

[0276] To generate sequences that fold into the beta solenoid structure, ProteinMPNN was used as described in Example 1 above. In most cases the repetitive pattern of beta strands was enforced 127

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[0834] by providing ProteinMPNN with specifications to make certain positions always have the same amino acid identity. Resulting sequences were validated in silico using Colabfold for structure prediction as described above.

[0835]

[0277] Once a Colabfold-based model was produced for each candidate sequence, the per-position metric pLDDT was used to rank designs. Designs with high average pLDDT (>70) were manually inspected to ensure that the high-confidence model had a beta solenoid structure and were identified as BSRPs with a tertiary beta solenoid structure. In some instances, linkers from a prespecified set of linker sequences, for example, any one or more of SEQ ID NOs: 122-125, 128-139, and 146, were inserted in the one or more loops connecting the beta strands of the beta solenoid polypeptides.

[0836]

[0278] The resulting sequences were validated in silico using pLDDT values as described above. In a subset of cases, the resulting BSRPs were used as backbone input to ProteinMPNN. In some instances, the linkers of the BSRPs were changed while retaining the same beta strands to generate new BSRPs. The amino acid sequences of the beta strand reservoir polypeptides (BSRPs) exhibiting a beta solenoid structure based on in silico analysis are set forth in Table 2. The DSSP (Define Secondary Structure of Proteins) program was used to identify the beta strands within each of these BSRPs.

[0837]

[0279] Additionally, an N-terminal methionine was added for protein expression if not already present and, in some cases - (i) one or two ‘GW’ or ‘W’ peptides were added to the C-terminus to improve the ability to quantify the protein via 280nm light absorbance; (ii) a ‘ SWTWENGKWTWK’ peptide (SEQ ID NO: 164) was added to the C-terminus to improve the ability to quantify the protein via 280nm light absorbance; (iii) a tag was added to the N- or C-terminus, together with a short linker, and / or (iv) a tag was added to the C-terminus, together with flanking short linkers (GSGS (SEQ ID NO: 167) or GGGSGGGS (SEQ ID NO: 1145)).

[0838] Example 3: Expression and Purification of Beta strand reservoir polypeptides

[0839]

[0280] The 82 BSRPs listed in Tables 1 and 2 and SEQ ID NO 15 and 84 were encoded in nucleic acid sequences SEQ ID NOs: 1061-1142, 2848 and 2849. Host cells were engineered to express each of the 83 BSRPs fused to an N-terminal ‘MHHHHHHGSGS (SEQ ID NO: 1144), C-terminal ‘GSGSHHHHHH’ (His-tag; SEQ ID NO: 161), and / or other tags, including those in 128

[0840] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0841] SEQ ID NO: 159-173 as detailed in Table 11 A. Due to the repetitive nature of the target DNA sequences, modular cloning strategies, such as Golden Gate assembly, were used. The polypeptides were purified using metal affinity chromatography (IMAC) purification. In some cases, the protein purification process involved the use of acids such as formic acid and acetic acid to prevent aggregation of the target proteins.

[0842]

[0281] Table 11 lists a subset of amino acid sequences disclosed herein:

[0843] Table 11:

[0844] SEQ ID Description Sequence

[0845] NO:

[0846] 172 N-terminal tag M[XorG]

[0847] 568 Dihedral beta strand TET

[0848] 573 Dihedral beta strand TKA

[0849] 621 Solenoid beta strand ENE

[0850] 627 Solenoid beta strand VEL

[0851] 635 Solenoid beta strand IEI

[0852] 643 Solenoid beta strand KNK

[0853] 667 Solenoid beta strand AKS

[0854] 794 Solenoid beta strand KAK

[0855] 817 Solenoid beta strand IKV

[0856] 840 Solenoid beta strand TEV

[0857] 882 Solenoid beta strand VEV

[0858] 999 Solenoid beta strand EAK

[0859] 1053 Solenoid beta strand EIE

[0860] 2072 Solenoid beta strand AGQ

[0861] 2084 Solenoid beta strand GAG

[0862] 2103 Solenoid beta strand GGY

[0863] 2121 Solenoid beta strand GQY

[0864] 2122 Solenoid beta strand GSQ

[0865] 2151 Solenoid beta strand QGA

[0866] 2172 Solenoid beta strand YGG

[0867] 2184 strands detected by DSSP4 KTL

[0868] secondary structure analysis

[0869] on the C6 flower model

[0870] 2193 strands detected by DSSP4 VKI

[0871] secondary structure analysis

[0872] on the C6 flower model

[0873] 2353 connectors between hairpins GEA

[0874]

[0875] 129

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[0877] 2558 connectors between hairpins TD

[0878] 2660 strand solenoid type 2 IKK

[0879] 2686 Strand solenoid type 4 KVE

[0880] 2725 Strand solenoid type 5 KVT

[0881] 2759 strand solenoid type 6 TVL

[0882]

[0883] Table 11 A

[0884]

[0282] Table 11A lists the DNA sequences encoding the BSRPs.

[0885] In-frame fusions of each of the BSRPs with one or DNA SEQ Design pA

[0886] more tags as represented by their SEQ ID NOs.

[0887] ID NO: ID

[0888] encoded by the respective DNA sequence

[0889] 1061 pA55724 < SEQ ID NO: 1>< SEQ ID NO: 161>

[0890] < SEQ ID NO: 172>< SEQ ID NO: 2>< SEQ ID NO:

[0891] 1062 pA55737

[0892] 161>

[0893] < SEQ ID NO: 163>< SEQ ID NO: 3>< SEQ ID NO:

[0894] 1063 pA55743

[0895] 159>

[0896] < SEQ ID NO: 4>< SEQ ID NO: 169>< SEQ ID NO:

[0897] 1064 pA56608

[0898] 161>

[0899] < SEQ ID NO: 172>< SEQ ID NO: 5>< SEQ ID NO:

[0900] 1065 pA56609

[0901] 167>< SEQ ID NO: 165>

[0902] < SEQ ID NO: 172XSEQ ID NO: 6XSEQ ID NO:

[0903] 1066 pA56610

[0904] 161>

[0905] < SEQ ID NO: 172XSEQ ID NO: 7XSEQ ID NO:

[0906] 1067 pA56612

[0907] 161>

[0908] < SEQ ID NO: 172XSEQ ID NO: 8XSEQ ID NO:

[0909] 1068 pA56613

[0910] 161>

[0911]

[0912] 130

[0913] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0914] < SEQ ID NO: 172>< SEQ ID NO: 9>< SEQ ID NO:

[0915] 1069 pA56614

[0916] 161>

[0917] 1070 pA56617 < SEQ IDNO: 10>< SEQ IDNO: 161>

[0918] 1071 pA56621 < SEQ IDNO: 11>< SEQ IDNO: 161>

[0919] 1072 pA56622 < SEQ IDNO: 12XSEQ IDNO: 161>

[0920] 1073 pA56624 < SEQ IDNO: 13XSEQ IDNO: 161>

[0921] 1074 pA56628 < SEQ IDNO: 14XSEQ IDNO: 161>

[0922] < SEQ ID NO: 15XSEQ ID NO: 169XSEQ ID NO:

[0923] 1075 pA56629

[0924] 161>

[0925] < SEQ ID NO: 16XSEQ ID NO: 169XSEQ ID NO:

[0926] 1076 pA56631

[0927] 161>

[0928] 1077 pA56633 < SEQ IDNO: 17XSEQ IDNO: 161>

[0929] 1078 pA56634 < SEQ IDNO: 18XSEQ IDNO: 161>

[0930] < SEQ ID NO: 19XSEQ ID NO: 169XSEQ ID NO:

[0931] 1079 pA56635

[0932] 161>

[0933] 1080 pA56636 < SEQ ID NO: 20XSEQ ID NO: 161>

[0934] 1081 pA56637 < SEQ ID NO: 21XSEQ ID NO: 161>

[0935] 1082 pA56638 < SEQ ID NO: 22XSEQ ID NO: 161>

[0936] 1083 pA56639 < SEQ ID NO: 23XSEQ ID NO: 161>

[0937] 1084 pA56640 < SEQ ID NO: 24XSEQ ID NO: 161>

[0938] < SEQ ID NO: 25XSEQ ID NO: 169XSEQ ID NO:

[0939] 1085 pA56641

[0940] 161>

[0941] 1086 pA56642 < SEQ ID NO: 26XSEQ ID NO: 161>

[0942]

[0943] 131

[0944] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0945] < SEQ ID NO: 172>< SEQ ID NO: 27>< SEQ ID NO:

[0946] 1087 pA56653

[0947] 161>

[0948] 1088 pA56654 < SEQ ID NO: 28>< SEQ ID NO: 161>

[0949] 1089 pA56657 < SEQ ID NO: 29>< SEQ ID NO: 161>

[0950] 1090 pA56660 < SEQ ID NO: 30>< SEQ ID NO: 161>

[0951] 1091 pA56662 < SEQ IDNO: 31>< SEQ IDNO: 161>

[0952] 1092 pA56663 < SEQ ID NO: 32>< SEQ ID NO: 161>

[0953] 1093 pA56664 < SEQ ID NO: 33>< SEQ ID NO: 161>

[0954] 1094 pA56665 < SEQ ID NO: 34>< SEQ ID NO: 161>

[0955] < SEQ ID NO: 35>< SEQ ID NO: 167>< SEQ ID NO:

[0956] 1095 pA56666

[0957] 165>

[0958] 1096 pA56667 < SEQ ID NO: 36>< SEQ ID NO: 161>

[0959] < SEQ ID NO: 37>< SEQ ID NO: 167>< SEQ ID NO:

[0960] 1097 pA56668

[0961] 165>

[0962] 1098 pA56669 < SEQ ID NO: 38>< SEQ ID NO: 161>

[0963] 1099 pA56673 < SEQ ID NO: 39>< SEQ ID NO: 161>

[0964] 1100 pA56674 < SEQ ID NO: 40>< SEQ ID NO: 161>

[0965] 1101 pA56676 < SEQ ID NO: 41>< SEQ ID NO: 161>

[0966] 1102 pA56677 < SEQ ID NO: 42>< SEQ ID NO: 161>

[0967] 1103 pA56678 < SEQ ID NO: 43>< SEQ ID NO: 161>

[0968] 1104 pA56679 < SEQ ID NO: 44>< SEQ ID NO: 161>

[0969] 1105 pA56680 < SEQ ID NO: 45>< SEQ ID NO: 161>

[0970] < SEQ ID NO: 172>< SEQ ID NO: 46>< SEQ ID NO:

[0971] 1106 pA56733

[0972] 161>

[0973]

[0974] 132

[0975] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[0976] < SEQ ID NO: 172>< SEQ ID NO: 47>< SEQ ID NO:

[0977] 1107 pA56735

[0978] 161>

[0979] < SEQ ID NO: 172>< SEQ ID NO: 48>< SEQ ID NO:

[0980] 1108 pA56736

[0981] 161>

[0982] < SEQ ID NO: 172>< SEQ ID NO: 49>< SEQ ID NO:

[0983] 1109 pA56737

[0984] 161>

[0985] < SEQ ID NO: 172>< SEQ ID NO: 50>< SEQ ID NO:

[0986] 1110 pA56738

[0987] 161>

[0988] < SEQ ID NO: 172>< SEQ ID NO: 51>< SEQ ID NO: mi pA56739

[0989] 161>

[0990] < SEQ ID NO: 172>< SEQ ID NO: 52>< SEQ ID NO:

[0991] 1112 pA56741

[0992] 161>

[0993] < SEQ ID NO: 172>< SEQ ID NO: 53>< SEQ ID NO:

[0994] 1113 pA56742

[0995] 161>

[0996] 1114 pA56756 < SEQ ID NO: 54>< SEQ ID NO: 161>

[0997] 1115 pA56775 < SEQ IDNO: 55>< SEQ IDNO: 161>

[0998] 1116 pA56776 < SEQ IDNO: 56>< SEQ IDNO: 161>

[0999] 1117 pA56777 < SEQ IDNO: 57>< SEQ IDNO: 161>

[1000] 1118 pA56778 < SEQ IDNO: 58>< SEQ IDNO: 161>

[1001] 1119 pA56779 < SEQ IDNO: 59>< SEQ IDNO: 161>

[1002] 1120 pA56780 < SEQ ID NO: 60>< SEQ ID NO: 161>

[1003] < SEQ ID NO: 61>< SEQ ID NO: 169>< SEQ ID NO:

[1004] 1121 pA56782

[1005] 161>

[1006] < SEQ ID NO: 62>< SEQ ID NO: 169>< SEQ ID NO:

[1007] 1122 pA57141

[1008] 160>

[1009]

[1010] 133

[1011] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1012] < SEQ ID NO: 162>< SEQ ID NO: 63>< SEQ ID NO:

[1013] 1123 pA57186

[1014] 161>

[1015] < SEQ ID NO: 172>< SEQ ID NO: 64>< SEQ ID NO:

[1016] 1124 pA57187

[1017] 169>< SEQ ID NO: 160>

[1018] < SEQ ID NO: 65>< SEQ ID NO: 169>< SEQ ID NO:

[1019] 1125 pA57193

[1020] 160>

[1021] < SEQ ID NO: 162>< SEQ ID NO: 66>< SEQ ID NO:

[1022] 1126 pA57250

[1023] 161>

[1024] < SEQ ID NO: 162>< SEQ ID NO: 67>< SEQ ID NO:

[1025] 1127 pA57251

[1026] 161>

[1027] < SEQ ID NO: 162>< SEQ ID NO: 68>< SEQ ID NO:

[1028] 1128 pA57252

[1029] 161>

[1030] < SEQ ID NO: 162>< SEQ ID NO: 69>< SEQ ID NO:

[1031] 1129 pA57253

[1032] 161>

[1033] < SEQ ID NO: 162>< SEQ ID NO: 70>< SEQ ID NO:

[1034] 1130 pA57254

[1035] 161>

[1036] < SEQ ID NO: 162>< SEQ ID NO: 71>< SEQ ID NO:

[1037] 1131 pA57255

[1038] 161>

[1039] < SEQ ID NO: 162>< SEQ ID NO: 72>< SEQ ID NO:

[1040] 1132 pA57256

[1041] 161>

[1042] < SEQ ID NO: 162>< SEQ ID NO: 73>< SEQ ID NO:

[1043] 1133 pA57259

[1044] 161>

[1045] < SEQ ID NO: 162>< SEQ ID NO: 74>< SEQ ID NO:

[1046] 1134 pA57260

[1047] 161>

[1048] < SEQ ID NO: 162>< SEQ ID NO: 75>< SEQ ID NO:

[1049] 1135 pA57474

[1050] 161>

[1051]

[1052] 134

[1053] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1054] < SEQ ID NO: 76>< SEQ ID NO: 169>< SEQ ID NO:

[1055] 1136 pA57501

[1056] 160>

[1057] < SEQ ID NO: 77>< SEQ ID NO: 169>< SEQ ID NO:

[1058] 1137 pA57503

[1059] 160>

[1060] < SEQ ID NO: 162>< SEQ ID NO: 78>< SEQ ID NO:

[1061] 1138 pA57506

[1062] 169>< SEQ ID NO: 161>

[1063] < SEQ ID NO: 172>< SEQ ID NO: 79>< SEQ ID NO:

[1064] 1139 pA57536

[1065] 160>

[1066] < SEQ ID NO: 172>< SEQ ID NO: 80>< SEQ ID NO:

[1067] 1140 pA60317

[1068] 161>

[1069] < SEQ ID NO: 172>< SEQ ID NO: 81>< SEQ ID NO:

[1070] 1141 pA60319

[1071] 161>

[1072] < SEQ ID NO: 172>< SEQ ID NO: 82>< SEQ ID NO:

[1073] 1142 pA60321

[1074] 161>

[1075] < SEQ ID NO: 172>< SEQ ID NO: 83>< SEQ ID NO:

[1076] 2848 pA60320

[1077] 161>

[1078] < SEQ ID NO: 172>< SEQ ID NO: 84>< SEQ ID NO:

[1079] 2849 pA61868

[1080] 161>

[1081]

[1082] Example 3A: Variations of the BSRPs

[1083]

[0283] The BSRPs listed in Table 1 and Table 2 (except for SEQ ID NO 83) and SEQ ID NO: 15 comprise one or more of the linkers of the amino acid sequence of any one or more of SEQ ID NOs: 122-125, 128-139, 146 and 1775-2063. Each of the following sequences of one or more of SEQ ID NOs: 122-158 when present in the BSRP was replaced with one of the linkers of the amino acid sequence of any one of SEQ ID NOs: 175-194, generating 1476 additional potential BSRP sequences (82 BSRPs times 18 sequences of SEQ ID NOs: 175-194). The structure of each of the 1476 potential BSRP sequences was predicted using Colabfold. The sequences that yielded 135

[1084] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1085] Colabfold structures with an average pLDDT of >70 and those that had a structure similar to the original BSRPs comprising any one of SEQ ID NOs: 122-125, 128-139, 146 and 1775-2063 were selected and are listed in Table 4. Exemplary structures of the BSRPs listed in Table 4 below are shown in FIGs. 9A-9D. In some sequences, an N-terminal methionine, or a C-terminal GSGS linker was included in the sequence. The inclusion of the methionine or GSGS linker does not alter the general conclusions of this analysis.

[1086] 136

[1087] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1088] Table 4:

[1089] BSRP SEQID Sequence Average name NO: pLDDT pA56775_l 195 MNKIKKTKNSEVKSTGENNEIESKSKKGKNKIESTVPAFGGNENKIESTGDNNKIESTIPGVGGNKNKIESTGDKNKIESTVPAVGGN 902 ENKIESKGNENKIESTIPAWGGNKNKIESKGDKNKIESTIPGLGGNENKIESKGDKNKIESKVPGIGGNKNKIESKGNKNKIESKVPGIG GNKNKIESKGNKNKIESKIPAYGGGENKIESKGNENKIESKWAVGGNKNKIESTGNKNKIESEGSGEIKVSGSGEITSNGNVNVTKK

[1090] pA55743_l 196 ALKYTIE VTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKAS AS AASRSQSASQSQALKYTIEVTESGGKAEYW 742 VKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASAASRSQSASQSQ ALKYTIE VTESGGKAEYVWKDNTGAILKAQKISNGDE VTVTTASGLKITVKASASAASRSQSASQSQALKYTIEVTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASA ASRSQSASQSQALKYTIEVTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASAASRSQSASQSQALKYTIEVT ESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASAASRSQSASQSQALKYTIEVTESGGKAEYVWKDNTGAIL KAQKISNGDEVTVTTASGLKITVKASASAASRSQSASQSQALKYTIEVTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGL KITVKASA

[1091] pA55743_2 197 ALKYTIE VTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASYPSSQYQTPSYPSSQYQTPSYPSSQYQTPALK 785 YTIEVTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASYPSSQYQTPSYPSSQYQTPSYPSSQYQTP ALKYTIE VTESGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVK AS ASYPSSQYQTPSYPSSQYQTPSYPSSQYQTP ALKYTIE VTE SGGKAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASYPSSQYQTPSYPSSQYQTPSYPSSQYQTPALKYTIEVTESGG KAEYVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASYPSSQYQTPSYPSSQYQTPSYPSSQYQTPALKYTIEVTESGGKAE YVWKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASYPSSQYQTPSYPSSQYQTPSYPSSQYQTPALKYTIEVTESGGKAEYW VKDNTGAILKAQKISNGDEVTVTTASGLKITVKASASYPSSQYQTPSYPSSQYQTPSYPSSQYQTPALKYTIEVTESGGKAEYVWKD NTGAILKAQKISNGDEVTVTTASGLKITVKASA

[1092] pA56608_l 198 MEIETEGKVEVELEGEAEAKVKLEGKNNEAKVKLEGEVPAVGKGEAKVKLEGDKNEAKVKLEGKVPAAGENEAKVKLEGNKNEA 907 KVKLEGDIPALGENEAKVKLEGNKNEAKVKLEGKIPGMGKNEAKVKLEGNKNEAKVKLEADVPGVGESEAKVKLEGNKNEAKVK LEGNVPAVGKSEAKVKLEGNKNEAKVKLEGNIPGVGESEAKVKLEGGKNEAKVKLEGNIPGAGEAEAKVKLEGNKNEAKVKLEG WPAVGEAEAKVKLEGNKNEAKVKLEGWPAAGEAEAKVKLEGNKNEAKVKLEGNIPAMGEAEAKVKLEGNNNEAKVKLEGNV PAYGEAEAKVKLEGNNNEAKVKLEGNVPAVGENEAKVKLEGNNQEAKVKLEGKIPGAGENEAKVKLEGNNQEAKVKLEGEVPA MGKNEAKVKLEGKNQEAKVKLEGKVPAGGENEAKVKLEGENNEAKVKLEGDVPGVGKGEAKVKLEGKNQEAKVKLEKGKKAE AEKKNEKVELKW

[1093]

[1094] 137

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[1096] pA56608_2 199 MEIETEGKVEVELEGEAEAKVKLEGKNNEAKVKLEGEIPGNGKGEAKVKLEGDKNEAKVKLEGKIPGRGENEAKVKLEGNKNEAK 910

[1097] VKLEGDVPGKGENEAKVKLEGNKNEAKVKLEGKVPGEGKNEAKVKLEGNKNEAKVKLEADIPATGESEAKVKLEGNKNEAKVKL EGNVPASGKSEAKVKLEGNKNEAKVKLEGNIPGNGESEAKVKLEGGKNEAKVKLEGNIPGEGEAEAKVKLEGNKNEAKVKLEGN VPAQGEAEAKVKLEGNKNEAKVKLEGNVPARGEAEAKVKLEGNKNEAKVKLEGNIPGDGEAEAKVKLEGNNNEAKVKLEGNVP ATGEAEAKVKLEGNNNEAKVKLEGNVPGEGENEAKVKLEGNNQEAKVKLEGKIPAKGENEAKVKLEGNNQEAKVKLEGEVPGEG KNEAKVKLEGKNQEAKVKLEGKIPGTGENEAKVKLEGENNEAKVKLEGDIPATGKGEAKVKLEGKNQEAKVKLEKGKKAEAEKK NEKVELKW

[1098] pA56609_l 200 MAEIKVKNSENLKIKSEGNTKEELESKGENNKEELESKGNIPAVGNNKLELESKGENNKLELESKGNVPAMGNNKLELESKGENNK 894

[1099] LELESKGNIPGVGDNKEELESKGENNKLELESKGNVPAVGENKLELESKGENNKLELESKANIPAVGENKLELESKGENNKLELESK GNVPAWGENKLELESKGEENKLELESKGDVPGGGETKLELESKGEKNKLELESKGNIPGVGETKLELESKGEENKLELESKGDVPG YGKTKLELESKGEKNKLELESKGDVPAVGESKLELESKGEKNKLELESKGKIPGGGNTKLELESKGEENKLELESKGDVPGVGNSKL ELESKGEKNKLELESKADVPGVGNSKLELESKGDENKLELESKADGTIKTKATEGGKGKAEVGSGS

[1100] pA56609_2 201 MAEIKVKNSENLKIKSEGNTKLELESKGENNKLELESKGNWGNGNNKLELESKGENNKLELESKGNIPAEGNNKLELESKGENNKL 907

[1101] ELESKGNWANGDNKLELESKGENNKLELESKGNWANGENKLELESKGENNKLELESKANIPAKGENKLELESKGENNKLELESK GNIPADGENKLELESKGEENKLELESKGDVPAEGETKLELESKGEKNKLELESKGNIPAHGETKLELESKGEENKLELESKGDIPGNG KTKLELESKGEKNKLELESKGDVPAEGESKLELESKGEKNKLELESKGKVPADGNTKLELESKGEENKLELESKGDVPGDGNSKLEL ESKGEKNKLELESKADIPGHGNSKLELESKGDENKLELESKADGTIKTKATEGGKGKAEVGSGS

[1102] pA56610_l 202 MAKIEIKNYENLEIESEGKTELKIELKGEKNKAKAKSEGKAAKDVKFGADARALENELKIELKGNKNKAKAKSEGEAAKDVKFGAD 88 1

[1103] ARALKNELKIELKGNKNKAKAKSEGDAAKDVKFGADARALKNELKIELKGNKNKAKAKSEGEAAKDVKFGADARALKTELKIEL KGNKNKAKAKSEADAAKDVKFGADARALKTELKIELKGNKNKAKAKSEGDAAKDVKFGADARALKTELKIELKGNKNKAKAKS EGKAAKDVKFGADARALKTELKIELKGGKNKAKAKSEGDAAKDVKFGADARALKTELKIELKGNKNKAKAKSEGDAAKDVKFG ADARALKTELKIELKGNKNKAKAKSEGDAAKDVKFGADARALKSELKIELKGNKNKAKAKSEGDAAKDVKFGADARALNSELKI ELKGNKNKAKAKSEGKAAKDVKFGADARALNSELKIELKGNKNKAKAKSEADAAKDVKFGADARALNSELKIELKGDKNKAKA KSEKSGKIEKEATEGAKVEAKV

[1104] pA56610_2 203 MAKIEIKNYENLEIESEGKTELKIELKGEKNKAKAKSEGKVPGVGENELKIELKGNKNKAKAKSEGEVPAGGKNELKIELKGNKNK 908

[1105] AKAKSEGDVPGLGKNELKIELKGNKNKAKAKSEGEIPAVGKTELKIELKGNKNKAKAKSEADVPAVGKTELKIELKGNKNKAKAK SEGDIPAGGKTELKIELKGNKNKAKAKSEGKVPAVGKTELKIELKGGKNKAKAKSEGDIPALGKTELKIELKGNKNKAKAKSEGDV PGVGKTELKIELKGNKNKAKAKSEGDVPAAGKSELKIELKGNKNKAKAKSEGDIPGMGNSELKIELKGNKNKAKAKSEGKVPAGG NSELKIELKGNKNKAKAKSEADVPGWGNSELKIELKGDKNKAKAKSEKSGKIEKEATEGAKVEAKV

[1106]

[1107] 138

[1108] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1109] pA56610_3 204 MAKIEIKNYENLEIESEGKTELKIELKGEKNKAKAKSEGKIPARGENELKIELKGNKNKAKAKSEGEIPGNGKNELKIELKGNKNKAK 917

[1110] AKSEGDVPAQGKNELKIELKGNKNKAKAKSEGEIPGSGKTELKIELKGNKNKAKAKSEADIPGTGKTELKIELKGNKNKAKAKSEG DVPGTGKTELKIELKGNKNKAKAKSEGKIPGHGKTELKIELKGGKNKAKAKSEGDVPGQGKTELKIELKGNKNKAKAKSEGDIPAQ GKTELKIELKGNKNKAKAKSEGDIPAHGKSELKIELKGNKNKAKAKSEGDIPGDGNSELKIELKGNKNKAKAKSEGKIPATGNSELK IELKGNKNKAKAKSEADIPAEGNSELKIELKGDKNKAKAKSEKSGKIEKEATEGAKVEAKV

[1111] pA56612_l 205 MAKIEIKNYENLEIESEGKTELKIELKGEIPARGKNKAKAKSEGKENELKIELKGNIPGNGKNKAKAKSEGEKNELKIELKGNIPASGK 91 8

[1112] NKAKAKSEGDKNELKIELKGNVPGTGKNKAKAKSEGEKTELKIELKGNIPGEGKNKAKAKSEADKTELKIELKGNIPGRGKNKAKA KSEGDKTELKIELKGNIPATGKNKAKAKSEGKKTELKIELKGGVPAEGKNKAKAKSEGDKTELKIELKGNIPGHGKNKAKAKSEGD KTELKIELKGNVPAKGKNKAKAKSEGDKSELKIELKGNIPGDGKNKAKAKSEGDNSELKIELKGNIPAHGKNKAKAKSEGKNSELKI ELKGNVPANGKNKAKAKSEADNSELKIELKGDIPGQGKNKAKAKSEKSGKIEKEATEGAKVEAKV

[1113] pA56621_l 206 MEIETEGKVEVELEGEAEAKVKLEGKGRGDSPYSGRGDSPYSNNEAKVKLEGEKGEAKVKLEGDGRGDSPYSGRGDSPYSKNEAK 85 1

[1114] VKLEGKENEAKVKLEGNGRGDSPYSGRGDSPYSKNEAKVKLEGDENEAKVKLEGNGRGDSPYSGRGDSPYSKNEAKVKLEGKKN EAKVKLEGNGRGDSPYSGRGDSPYSKNEAKVKLEADESEAKVKLEGNGRGDSPYSGRGDSPYSKNEAKVKLEGNKSEAKVKLEGN GRGDSPYSGRGDSPYSKNEAKVKLEGNESEAKVKLEGGGRGDSPYSGRGDSPYSKNEAKVKLEGNEAEAKVKLEGNGRGDSPYSG RGDSPYSKNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSKNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSKN EAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSNNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSNNEAKVKLEG NENEAKVKLEGNGRGDSPYSGRGDSPYSNQEAKVKLEGKENEAKVKLEGNGRGDSPYSGRGDSPYSNQEAKVKLEGEKNEAKVK LEGKGRGDSPYSGRGDSPYSNQEAKVKLEGKENEAKVKLEGEGRGDSPYSGRGDSPYSNNEAKVKLEGDKGEAKVKLEGKGRGD SPYSGRGDSPYSNQEAKVKLEKGKKAEAEKKNEKVELK

[1115] pA56621_2 207 MEIETEGKVEVELEGEAEAKVKLEGKGRGDSPYSGRGDSPYSGRGDSPYSNNEAKVKLEGEKGEAKVKLEGDGRGDSPYSGRGDS 73 8

[1116] PYSGRGDSPYSKNEAKVKLEGKENEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVKLEGDENEAKVKLEGNGRGDSP YSGRGDSPYSGRGDSPYSKNEAKVKLEGKKNEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVKLEADESEAKVKLEG NGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVKLEGNKSEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVKLEGNESE AKVKLEGGGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVK LEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSKNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYS KNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSNNEAKVKLEGNEAEAKVKLEGNGRGDSPYSGRGDSPYSG RGDSPYSNNEAKVKLEGNENEAKVKLEGNGRGDSPYSGRGDSPYSGRGDSPYSNQEAKVKLEGKENEAKVKLEGNGRGDSPYSGR GDSPYSGRGDSPYSNQEAKVKLEGEKNEAKVKLEGKGRGDSPYSGRGDSPYSGRGDSPYSNQEAKVKLEGKENEAKVKLEGEGRG DSPYSGRGDSPYSGRGDSPYSNNEAKVKLEGDKGEAKVKLEGKGRGDSPYSGRGDSPYSGRGDSPYSNQEAKVKLEKGKKAEAEK KNEKVELK

[1117]

[1118] 139

[1119] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1120] pA56621_3 MEIETEGKVEVELEGEAEAKVKLEGKSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPNNEAKVKLEGEKGEAKVKLEGDSYPSSQYQT 845

[1121] PSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGKENEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGDENE AKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGKKNEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQY QTPKNEAKVKLEADESEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGNKSEAKVKLEGNSYPSSQYQTP SYPSSQYQTPSYPSSQYQTPKNEAKVKLEGNESEAKVKLEGGSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGNEAEA KVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGNEAEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQT PKNEAKVKLEGNEAEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNEAKVKLEGNEAEAKVKLEGNSYPSSQYQTPS YPSSQYQTPSYPSSQYQTPNNEAKVKLEGNEAEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPNNEAKVKLEGNENEAK VKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPNQEAKVKLEGKENEAKVKLEGNSYPSSQYQTPSYPSSQYQTPSYPSSQYQTP NQEAKVKLEGEKNEAKVKLEGKSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPNQEAKVKLEGKENEAKVKLEGESYPSSQYQTPSYP SSQYQTPSYPSSQYQTPNNEAKVKLEGDKGEAKVKLEGKSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPNQEAKVKLEKGKKAEAE KKNEKVELK

[1122] pA56628_l 209 MVKVEGEGVKVKVEGEAKAEAKAEGEVPGVGGNKIEIKLEGKKNKAEAKAEGDVPGVGGNKIEIKLEGEENKAEAKAEGEVPAM 860

[1123] GGNKIEIKLEGDKNKAEAKAEGEIPGIGGNKIEIKLEGEKNKAEAKAEGEIPAAGGNKIEIKLEGDEAKAEAKAEGEIPAGGGNKIEIK LEGSGKAKALPGETEVDDEGAKVKVEEKK

[1124] pA56666 1 210 MVKVEGEGVKVKVEGEAKAEAKAEGEGRGDSPYSGRGDSPYSGNKIEIKLEGKKNKAEAKAEGDGRGDSPYSGRGDSPYSGNKIEI 867

[1125] KLEGEENKAEAKAEGEGRGDSPYSGRGDSPYSGNKIEIKLEGDKNKAEAKAEGEGRGDSPYSGRGDSPYSGNKIEIKLEGEKNKAEA KAEGEGRGDSPYSGRGDSPYSGNKIEIKLEGDEAKAEAKAEGEGRGDSPYSGRGDSPYSGNKIEIKLEGSGKAKALPGETEVDDEGA KVKVEEKKGSGS

[1126] pA56629_l 211 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKIPATGKNKAEAKAEGDGNKIEIKLEGEVPGHGENKAEAKAEGEGNKIEI 85 5

[1127] KLEGDVPAKGKNKAEAKAEGEGNKIEIKLEGEVPGHGKNKAEAKAEGEGNKIEIKLEGDIPGNGEAKAEAKAEGEGNKIEIKLEGSG KAKALPGETEVDDEGAKVKVEEKKW

[1128] pA56631_l 86 1

[1129] pA56631_2 214 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKIPGNGKNKAEAKAEGDGNKIEIKLEGEIPAKGENKAEAKAEGEGNKIEI 842

[1130] KLEGDVPANGKNKAEAKAEGEGNKIEIKLEGEVPADGKNKAEAKAEGEGNKIEIKLEGDIPAKGEAKAEAKAEGEGNKIEIKLEGSG KAKALPGETEVDDEGAKVKVEEKKW

[1131]

[1132] 140

[1133] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246 pA56631_3 215 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKGRGDSPYSGRGDSPYSKNKAEAKAEGDGNKIEIKLEGEGRGDSPYSGR 867

[1134] GDSPYSENKAEAKAEGEGNKIEIKLEGDGRGDSPYSGRGDSPYSKNKAEAKAEGEGNKIEIKLEGEGRGDSPYSGRGDSPYSKNKAE AKAEGEGNKIEIKLEGDGRGDSPYSGRGDSPYSEAKAEAKAEGEGNKIEIKLEGSGKAKALPGETEVDDEGAKVKVEEKKW

[1135] pA56633_l 216 MMKIKKTENAEVKATGEIPALGNNEIEVESEKGKNKAELKGEIPALGKNKAELKGNNNKAELKGNVPAFGKNKAELKGNNNKAEL 905

[1136] KGNVPALGKNKAELKGNDNKAELKGNIPAVGKNKAELKGNKNKAELKGNVPGYGKNKAELKGNKNKAELKGNIPGMGKNKAEL KGNKNKAELKGNVPGVGKNKAELKGNKNKAELKGGVPAVGENKAELKGKKNKAELKGEIPAMGKNKAELKGNKNKAELTGKV PGMGTTEVTIKGGGEAYVKGEGKVEV

[1137] pA56639_l 217 MMKIKKTENAEVKATGENNEIEVESEKGKNKAELKGEKNKAELKGNIPAEGNNKAELKGNKNKAELKGNVPATGNNKAELKGNK 925

[1138] NKAELKGNVPGEGDNKAELKGNKNKAELKGNVPGEGKNKAELKGNKNKAELKGNVPAEGKNKAELKGNKNKAELKGNIPGEGK NKAELKGNKNKAELKGNIPGSGKNKAELKGGENKAELKGKVPGDGKNKAELKGEKNKAELKGNVPGNGKNKAELTGKTTEVTIK GGGEAYVKGEGKVEV

[1139] pA56680_l 218 MMKIKKTENAEVKATGENNEIEVESEKGKNKAELKGEKNKAELKGNGRGDSPYSGRGDSPYSNNKAELKGNKNKAELKGNGRGD 924

[1140] SPYSGRGDSPYSNNKAELKGNKNKAELKGNGRGDSPYSGRGDSPYSDNKAELKGNKNKAELKGNGRGDSPYSGRGDSPYSKNKAE LKGNKNKAELKGNGRGDSPYSGRGDSPYSKNKAELKGNKNKAELKGNGRGDSPYSGRGDSPYSKNKAELKGNKNKAELKGNGR GDSPYSGRGDSPYSKNKAELKGGENKAELKGKGRGDSPYSGRGDSPYSKNKAELKGEKNKAELKGNGRGDSPYSGRGDSPYSKNK AELTGKTTEVTIKGGGEAYVKGEGKVEV

[1141] pA56669 1 219 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKIPGFGKNKAEAKAEGDGNKIEIKLEGEIPALGENKAEAKAEGEGNKIEIK 882

[1142] LEGDVPGFGKNKAEAKAEGEGNKIEIKLEGEVPAWGKNKAEAKAEGEGNKIEIKLEGDVPGVGEAKAEAKAEGEGNKIEIKLEGSG KAKALPGETEVDDEGAKVKVEEKK

[1143] pA56669_2 220 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKIPAKGKNKAEAKAEGDGNKIEIKLEGEVPAQGENKAEAKAEGEGNKIEI 817

[1144] KLEGDVPADGKNKAEAKAEGEGNKIEIKLEGEVPGRGKNKAEAKAEGEGNKIEIKLEGDIPAEGEAKAEAKAEGEGNKIEIKLEGSG KAKALPGETEVDDEGAKVKVEEKK

[1145] pA56669_3 221 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKMDYPERYMDMSGWQMDKNKAEAKAEGDGNKIEIKLEGEMDYPEWF 71 1

[1146] MDMSGYQMDNQGRYMDENKAEAKAEGEGNKIEIKLEGDMDMSGWSMDTQGRYMDKNKAEAKAEGEGNKIEIKLEGEMDYPEW WMDYSNYSMDMHGRYMDKNKAEAKAEGEGNKIEIKLEGDMDYPEWFMDYSGYSMDEAKAEAKAEGEGNKIEIKLEGSGKAKAL PGETEVDDEGAKVKVEEKK

[1147] pA56669_4 222 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKSYPSSQYQTPSYPSSQYQTPKNKAEAKAEGDGNKIEIKLEGESYPSSQYQ 776

[1148] TPSYPSSQYQTPENKAEAKAEGEGNKIEIKLEGDSYPSSQYQTPSYPSSQYQTPKNKAEAKAEGEGNKIEIKLEGESYPSSQYQTPSYP

[1149]

[1150] 141

[1151] 331999525 v1Attorney Docket No.: ARZE-046 / 02WO 311023-2246

[1152] SSQYQTPKNKAEAKA. EGEGNKIEIKLEGDSYPSSQYQTPSYPSSQYQTPEAKAEAKAEGEGNKIEIKLEGSGKAKALPGETEVDDEG AKVKVEEKK

[1153] pA56669_5 223 MVKVEGEGVKVKVEGEAKAEAKAEGEGNKIEIKLEGKSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNKAEAKAEGDGNKIEIKLE 794

[1154] GESYPSSQYQTPSYPSSQYQTPSYPSSQYQTPENKAEAKAEGEGNKIEIKLEGDSYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNKAE AKAEGEGNKIEIKLEGESYPSSQYQTPSYPSSQYQTPSYPSSQYQTPKNKAEAKAEGEGNKIEIKLEGDSYPSSQYQTPSYPSSQYQTP SYPSSQYQTPEAKAEAKAEGEGNKIEIKLEGSGKAKALPGETEVDDEGAKVKVEEKK

[1155] pA56756_l 224 MFKVKAIGNTRAVIIGEGLLELVEPG...

Claims

Attorney Docket No.: ARZE-046 / 02WO 311023-2246CLAIMS1. A beta strand reservoir polypeptide, comprising two to twelve beta domains and a linker between each of the beta domains, wherein each of the beta domains comprises an amino acid sequence having at least about 75% identity to any one of SEQ ID NOs: 91-96, wherein the beta strand reservoir polypeptide optionally comprises eight or twelve beta domains.

2. The beta strand reservoir polypeptide of claim 1, wherein the beta domains associate with each other in silico to form an octameric structure having D4 dihedral symmetry or a dodecameric structure having D6 dihedral symmetry.

3. The beta strand reservoir polypeptide of claim 2, wherein each of the beta domains exhibits an average pLDDT of over 70.

4. A beta strand reservoir polypeptide, comprising eight or twelve beta domains and a linker between each of the beta domains, wherein:(i) the beta domains associate with each other in silico to form an octameric structure having D4 dihedral symmetry or a dodecameric structure having D6 dihedral symmetry, and(ii) each of the beta domains comprises about 40 amino acid residues to about 100 amino acid residues and exhibits an average pLDDT of over 70.

5. The beta strand reservoir polypeptide of claim 4, wherein each of the beta domains comprises an amino acid sequence having at least about 75% identity to any one of SEQ ID NOs: 91-96.265Attorney Docket No.: ARZE-046 / 02WO 311023-22466. The beta strand reservoir polypeptide of any one of claims 1-5, wherein each of the beta domains comprises about 40 amino acid residues to about 100 amino acid residues.

7. The beta strand reservoir polypeptide of any one of claims 1-6, wherein each of the beta domains comprises Lx-(S-L)y-Sz, wherein S is a beta strand and L is a linker, wherein x is 0 or 1, y is any one of 4 through 12, and z is 0 or 1, and wherein at least 30% of the amino acid residues of each of the beta domains are part of a beta strand.

8. The beta strand reservoir polypeptide of claim 7, wherein the beta strand comprises about 3 to about 10 amino acid residues.

9. The beta strand reservoir polypeptide of claim 7 or claim 8, wherein the beta strand comprises less than 50% glycine residues and / or less than 50% alanine residues.

10. The beta strand reservoir polypeptide of any one of claims 7-9, wherein the beta strand comprises an amino acid sequence of any one of SEQ ID NOs: 568-596.

11. The beta strand reservoir polypeptide of any one of claims 1-10, wherein the linker comprises about 5 to about 50 amino acid residues.

12. The beta strand reservoir polypeptide of any one of claims 1-11, wherein the linker comprises at least 20% glycine (G) residues.

13. The beta strand reservoir polypeptide of any one of claims 1-12, wherein the linker comprises at least 50% glycine (G) residues.

14. The beta strand reservoir polypeptide of any one of claims 1-13, wherein the linker and / or the beta domains do not comprise one or more of the following:(i) an amino acid sequence of (GA)nX, wherein X is any amino acid, and n>5,266Attorney Docket No.: ARZE-046 / 02WO 311023-2246(ii) an amino acid sequence of An, wherein n >5, and(iii) an amino acid sequence of (AG)n, wherein n >5.

15. The beta strand reservoir polypeptide of any one of claims 1-14, wherein the linker comprises an amino acid sequence having at least about 80% identity to any one of the amino acid sequences of SEQ IDNOs: 122-158, 175-194, 1143 and 1775-2063.

16. The beta strand reservoir polypeptide of any one of claims 1-15, wherein the beta strand reservoir polypeptide comprises an amino acid sequence having at least about 75% identity to an amino acid sequence of any one of SEQ ID NOs: 2, 3, 46-54, and 80-83.

17. A beta strand reservoir polypeptide, comprising at least four beta strands and a linker between each of the beta strands wherein the beta strand reservoir polypeptide comprises a beta solenoid motif, the beta solenoid motif comprising a consensus amino acid sequence of any one of SEQ ID NOs: 1590-1600, and 1602-1655, and optionally, wherein the at least four beta strands associate to form two parallel beta sheets (Bl and B2) in silico, and optionally, wherein the beta strand reservoir polypeptide comprises at least ten beta strands.

18. A beta strand reservoir polypeptide, comprising at least ten beta strands and a linker between each of the beta strands, wherein the at least ten beta strands associate to form two parallel beta sheets (Bl and B2), and wherein the polypeptide exhibits a beta solenoid structure in silico.

19. The beta strand reservoir polypeptide of any one of claims 17-18, wherein one or more Ca carbons of beta sheet Bl are separated by a distance in the range of about 6A to about 12A from one or more Ca carbons of beta sheet B2, and wherein beta strands in beta sheet Bl point in an opposite direction to those in beta sheet B2.267Attorney Docket No.: ARZE-046 / 02WO 311023-2246 20. The beta strand reservoir polypeptide of any one of claims 17-19, comprising:(L - S - L - S)x- L, wherein L is a loop, S is a beta strand, x > 2, and the L - S - L - S is about 17 to about 70 (e.g., about 18 to about 70) amino acids in length.

21. The beta strand reservoir polypeptide of any one of claims 17-20, wherein the beta strand reservoir polypeptide comprises one or more beta solenoid motifs having an amino acid sequence of any one of SEQ ID NOs: 1146-1259, 1261, and 1264-1589.

22. The beta strand reservoir polypeptide of any one of claims 17-21, wherein beta sheet Bl and / or beta sheet B2 comprises one or more beta strands having an amino acid sequence of any one of SEQ ID NOs: 597-787, 789-1053, 2064-2174 and 2850-2855.

23. The beta strand reservoir polypeptide of any one of claims 17-22, wherein the beta strand reservoir polypeptide comprises an amino acid sequence having at least about 75% identity to of any one of SEQ ID NOs: 1, 4-14, 16-45, and 55-79.

24. A beta strand reservoir polypeptide, comprising at least ten beta strands and a linker between each of the beta strands, wherein the at least ten beta strands associate to form two anti-parallel beta sheets (Bl and B2) in silico.

25. The beta strand reservoir polypeptide of claim 24, wherein the polypeptide comprises an amino acid sequence that is at least 75% identical the amino acid sequence of SEQ ID NO: 15.

26. A beta strand reservoir polypeptide comprising an amino acid sequence having at least about 75% identity to an amino acid sequence of any one of SEQ ID NOs: 1-84.268Attorney Docket No.: ARZE-046 / 02WO 311023-224627. A fusion protein, comprising a first amino acid sequence having at least about 75% identity to an amino acid sequence of any one of SEQ ID NOs: 1-84, and a second amino acid sequence.

28. The fusion protein of claim 27, wherein the second amino acid sequence comprises a protein tag.

29. The fusion protein of claim 27, wherein the second amino acid sequence comprises any one or more of SEQ ID NOs: 159-173, 1144 and 1145.

30. A non-naturally occurring beta stand reservoir protein (BSRP) containing an even number of at least 4 polypeptide stretches within its sequence wherein said polypeptide stretches have the following characteristics: (i) are all of the same length, (ii) between 40 and 100 amino acids in length, (iii) have a sequence identity of >75% for all possible pairwise comparisons, (iv) are connected by glycine-rich sequence segments (>30% glycine) that are 10 or more residues long, (vi) are less than 15% glycine, (vii) and wherein the expressed BSRP tests positive in the droplet drawing test.

31. A non-naturally occurring beta stand reservoir protein (BSRP) comprising a sequence of 200 or more amino acids, and having the following structural features (a) and (b) according to a beta solenoid structural model generated using a solenoid modeling protocol, wherein the BSRP comprises one or more beta strands identified by DSSP4 secondary structure analysis on the structural model: (a) the average pLDDT of all amino acid residues within a beta strand, is 60 or more, and (b) the majority (e.g. at least 70%, or at least, 75%, or at least 80%, or at least 82%, or at least 85%) of the amino acid residues that are part of the one or more beta strands are part of at most two beta sheets, and wherein the beta solenoid BSRP is positive in a droplet drawing test.269Attorney Docket No.: ARZE-046 / 02WO 311023-224632. An antiparallel beta solenoid BSRP, wherein the BSRP comprises one or more beta strands identified by DSSP4 secondary structure analysis on the structural model of the BSRP, wherein a majority (>50%) of amino acid residues are part of the one or more beta strands, and are included in two beta sheets that are both composed of antiparallel hairpins wherein the antiparallel beta solenoid BSRP passes a droplet drawing test.

33. A composition, comprising the beta strand reservoir polypeptide of any one of claims 1- 32.

34. A polynucleotide encoding the beta strand reservoir polypeptide of any one of claims 1- 32.

35. An expression cassette comprising the polynucleotide of claim 34.

36. A host cell comprising the beta strand reservoir polypeptide of any one of claims 1-32, the polynucleotide of claim 34, or the expression cassette of claim 35.

37. A method of preparing a beta strand reservoir polypeptide, the method comprising expressing the beta strand reservoir polypeptide of any one of claims 1-32 in a host cell or in a cell free expression system.

38. A lyophilized powder, comprising the beta strand reservoir polypeptide of any one of claims 1-32, or the polynucleotide of claim 34.

39. An in silico method of generating a beta strand reservoir polypeptide, comprising eight or twelve beta domains and a linker between each of the beta domains, wherein the beta domains associate with each other in silico to form an octameric structure having D4 dihedral symmetry or a dodecameric structure having D6 dihedral symmetry, the method comprising:270Attorney Docket No.: ARZE-046 / 02WO 311023-2246 (i) generating one or more amino acid sequences of a protein homodimer, wherein the protein homodimer comprises two protein monomers, each of which comprises greater than 25% beta strand secondary structure content,(ii) identifying one or more structural models of the protein homodimer, wherein each of the two protein monomers has an odd number of beta strands, wherein each of the beta strands comprises at least 3 amino acid residues, and wherein the first and last strands in primary sequence within each monomer each exhibit base pairing with two other strands in the homodimer.(iii) identifying one or more structural models of step (ii), wherein the protein homodimer exhibits C2 symmetry or C2 pseudo-symmetry,(iv) generating one or more of each of contact maps and secondary structure strings of the one or more structural models of step (iii),(v) generating one or more structural models of a C2 symmetric homodimer using the contact maps and / or the secondary structure strings from step (iv) as input, (vi) generating one or more structural models of a D4 multimer or a D6 multimer, wherein each D4 multimer comprises 8 protein monomers and each D6 multimer comprises 12 protein monomers using the models generated in step (v) as inputs, (vii) generating a sequence based on the backbone coordinates of the model output in step (vi), optionally using Protein MPNN,(viii) identifying one or more structural models having pLDDT > 70, and(ix) computationally linking the 8 or 12 protein monomers to generate the beta strand reservoir polypeptide.271Attorney Docket No.: ARZE-046 / 02WO 311023-224640. An in silico method of generating a beta strand reservoir polypeptide, comprising at least four beta strands and a linker between each of the beta strands, wherein the beta strand reservoir polypeptide comprises a beta solenoid motif, the method comprising:(i) generating a structural model of a protein backbone with beta solenoid structure, wherein the beta solenoid structure has at least four beta solenoid hairpins, (ii) generating one or more N-terminal and C-terminal backbone regions, wherein the backbone regions break the repetitive structural pattern of the solenoid hairpins, (iii) generating one or more amino acid sequences for the protein backbones generated in steps (i) and (ii),(iv) generating one or more structural models having pLDDT > 70, and(v) identifying one or more structural models exhibiting a beta solenoid structure.272

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