Polypeptides comprising beta-tricalcium phosphate-binding sequences and uses thereof

Abstract

Provided herein are polypeptides comprising one or more beta-tricalcium phosphate (beta TCP) binding sequences and uses thereof.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of October 11, 2019, application number 201980082730.1, and invention name “Polypeptides comprising β-tricalcium phosphate-binding sequences and uses thereof” (the application date of the corresponding PCT application is October 11, 2019, and the application number is PCT / US2019 / 055939).

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 744,941, filed on October 12, 2018; the entire contents of which are incorporated herein by reference. Technical Field

[0004] The present disclosure relates generally to the field of molecular biology and, more particularly, to methods of treating bone and cartilage loss defects in mammals. Background Art

[0005] An average of 6 million people break bones each year in the United States. Of these bone defects, the majority heal without problems. However, 5-10% will heal poorly or not at all (American Academy of Orthopaedic Surgeons). Large defects (also called critical-sized bone defects) may not heal spontaneously and may result in nonunion. Nonunion occurs when there is no sign of healing for at least three months and there is no hope of further healing (American Academy of Orthopaedic Surgeons). Currently, bone grafting is considered the "gold standard" for treating large or segmental bone defects. However, this treatment has limitations such as donor site pain, risk of rejection, limited donor supply, and risk of transmission of infectious diseases.

[0006] Repair of severe and complex bone and cartilage defects is also limited by poor tissue regrowth on implanted orthopedic substrates. Reasons for poor tissue regrowth include loss of implanted progenitor cells within 48 hours of implantation and progenitor cell deficiency. Another contributing factor is that current orthopedic substrates do not promote adequate tissue regeneration. Summary of the invention

[0007] The present invention is based on the following discovery: the use of a composition comprising beta-tricalcium phosphate (β-TCP) and a chimeric polypeptide can greatly improve bone healing and accelerate tissue re-growth, wherein the chimeric polypeptide comprises at least one β-TCP binding sequence and a mammalian growth factor (e.g., bone morphogenetic protein 2 (BMP-2)). Therefore, provided herein are chimeric polypeptides comprising one or more β-TCP binding sequences and a mammalian growth factor, compositions comprising any of these chimeric polypeptides (and optionally, β-TCP), and methods for promoting bone or cartilage formation, methods for replacing and / or repairing bone or cartilage, and methods for treating fractures or bone loss comprising administering any of these compositions. The compositions and methods provided herein can increase and maintain the number of progenitor cells at a site of bone and / or cartilage injury by stem cell capture. The compositions and methods provided herein can also be applied to local delivery of soft tissue repair or therapeutic agents.

[0008] The chimeric polypeptides provided herein include: (i) one or more β-tricalcium phosphate (βTCP)-binding sequences, wherein at least one of the one or more β-TCP-binding sequences is selected from the group consisting of: VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), TGSGDSSHHRPS (SEQ ID NO: 17), GSSGESTHHKPST (SEQ ID NO: 18), VGADSTHHRPVT (SEQ ID NO: 19), GAADTTHHRPVT (SEQ ID NO: 20), TGSGDSSHHRPS (SEQ ID NO: 21), GSSGESTHHKPST (SEQ ID NO: 22), VGADSTHHRPVT (SEQ ID NO: 23), GAADTTHHRPVT (SEQ ID NO: 24), TGSGDSSHHRPS (SEQ ID NO: 25), NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19); and (ii) mammalian growth factors.

[0009] In some embodiments, the chimeric polypeptide includes two or more β-TCP-binding sequences.

[0010] In some embodiments, each adjacent pair of the two or more β-TCP binding sequences are directly adjacent to each other.

[0011] In some embodiments, each adjacent pair of the two or more β-TCP binding sequences is separated by a linker sequence.

[0012] In some embodiments, the chimeric polypeptide includes at least two different β-TCP-binding sequences.

[0013] In some embodiments, the chimeric polypeptide includes two or more copies of the same β-TCP-binding sequence.

[0014] In some embodiments, at least one of the two or more β-TCP-binding sequences is LLADTTHHRPWT (SEQ ID NO: 1) or GQVLPTTTPSSP (SEQ ID NO: 19).

[0015] In some embodiments, the at least two different β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1) and VIGESTHHRPWS (SEQ ID NO: 2).

[0016] In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWS (SEQ ID NO: 20).

[0017] In some embodiments, the at least two different β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), and IIGESSHHKPFT (SEQ ID NO: 6).

[0018] In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFT (SEQ ID NO: 21).

[0019] In some embodiments, the at least two different β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), and ILAESTHHKPWT (SEQ ID NO: 4).

[0020] In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRPW GILAESTHHKPWT (SEQ ID NO: 22).

[0021] In some embodiments, the at least two β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1) and ILAESTHHKPWT (SEQ ID NO: 4).

[0022] In some embodiments, the chimeric polypeptide includes at least one copy of the sequence LLADTTHHRPWTILAESTHHKPWT (SEQ ID NO: 23).

[0023] In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTILAESTHHKPWTLLADTTHHRPWTILAESTHHKP WTLLADTTHHRPWT (SEQ ID NO: 24).

[0024] In some embodiments, the at least two β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1) and GLGDTTHHRPWG (SEQ ID NO: 7).

[0025] In some embodiments, the chimeric polypeptide includes at least one copy of the sequence of LLADTTHHRPWTGLGDTTHHRPWG (SEQ ID NO: 25).

[0026] In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 26).

[0027] In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHR PWGLLADTTHHRPWT (SEQ ID NO: 27).

[0028] In some embodiments, the chimeric polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHR PWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 28).

[0029] In some embodiments, the at least two β-TCP-binding sequences include STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), and GSSGESTHHKPST (SEQ ID NO: 14).

[0030] In some embodiments, the chimeric polypeptide comprises the sequence of STADTSHHRPSTSGGESTHHRPSTSGGESSHHKPSTGSGDSSHHRPSG SSGESTHHKPST (SEQ ID NO: 29).

[0031] In some embodiments, the at least two β-TCP-binding sequences include VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18), and GGADTTHHRPGT (SEQ ID NO: 19).

[0032] In some embodiments, the chimeric polypeptide comprises the sequence of VGADSTHHRPVTGAADTTHHRPVTAGADTTHHRPVTGGADTTHHR PATGGADTTHHRPGT (SEQ ID NO: 30).

[0033] In some embodiments, the at least two different β-TCP-binding sequences include STADTSHHRPS (SEQ ID NO: 10), LLADTTHHRPWT (SEQ ID NO: 1), TSGGESTHHRPS (SEQ ID NO: 11), VGADSTHHRPVT (SEQ ID NO: 15), TSGGESSHHKPS (SEQ ID NO: 12), GAADTTHHRPVT (SEQ ID NO: 16), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPS (SEQ ID NO: 14), and TGGADTTHHRPAT (SEQ ID NO: 18).

[0034] In some embodiments, the chimeric polypeptide comprises the sequence of STADTSHHRPSLLADTTHHRPWTTSGGESTHHRPSVGADSTHHRPVTTSGGESSHHKPSGAADTTHHRPVTTGSGDSSHHRPSGSSGESTHHKPSTGGADTTHHRPAT (SEQ ID NO: 31).

[0035] In some embodiments of any of the chimeric polypeptides described herein,The mammalian growth factor is selected from the group consisting of epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF-1), fibroblast growth factor (FGF), fibroblast growth factor 2 (FGF2), fibroblast growth factor 18 (FGF18), transforming growth factor α (TGF-α), transforming growth factor β (TGF-β), transforming growth factor β1 (TGF-β1), transforming growth factor β3 (TGF-β3), osteogenic protein 1 (OP-1), osteogenic protein 2 (OP-2), osteogenic protein 3 (OP-3), bone morphogenetic protein 2 (BMP-2), bone morphogenetic protein 3 (BMP-3), bone morphogenetic protein 4 ( BMP-4), bone morphogenetic protein 5 (BMP-5), bone morphogenetic protein 6 (BMP-6), bone morphogenetic protein 7 (BMP-7), bone morphogenetic protein (BMP-9), bone morphogenetic protein 10 (BMP-10), bone morphogenetic protein 11 (BMP-11), bone morphogenetic protein 12 (BMP-12), bone morphogenetic protein protein 13 (BMP-13), bone morphogenetic protein 15 (BMP-15), dentin phosphoprotein (DPP), plant-related growth factor (Vgr), growth differentiation factor 1 (GDF-1), growth differentiation factor 3 (GDF-3), growth differentiation factor 5 (GDF-5), growth differentiation factor 6 (GDF-6), Growth differentiation factor 7 (GDF-7), growth differentiation factor 8 (GDF8), growth differentiation factor 11 (GDF11), growth differentiation factor 15 (GDF15), vascular endothelial growth factor (VEGF), hyaluronan binding protein (HABP), collagen binding protein (CBP), fibroblast growth factor 18 (FGF-18), keratinocyte growth factor (KGF), tumor necrosis factor α (TNFα), tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), wnt family member 1 (WNT1), wnt family member 2 (WNT2), wnt family member 2B (WNT2B), wnt family member 3 (WNT3), wnt family Wnt family member 3A (WNT3A), wnt family member 4 (WNT4), wnt family member 5A (WNT5A), wnt family member 5B (WNT5B), wnt family member 6 (WNT6), wnt family member 7A (WNT7A), wnt family member 7B (WNT7B), wnt family member 8A (WNT8A), wnt family member 8B (WNT8B), wnt family member 9A (WNT9A), wnt family member 9B (WNT9B), wnt family member 10A (WNT10A), wnt family member 10B (WNT10B), wnt family member 11 (WNT11) and wnt family member 16 (WNT16).

[0036] In some embodiments, the mammalian growth factor is BMP-2.

[0037] In some embodiments, BMP-2 includes MVAGTRCLLALLLPQVLLGGAAGLVPELGRRKFAAASSGRPSSQPSDEVLSEFELRLLSMFGLKQRPTPSRDAVVPPYMLDLYRRHSG QPGSPAPDHRLERAASRANTVRSFHHEESLEELPETSGKTTRRFFFNLSSIPTEEFITSAELQVFREQMQDALGNNSSFHHRINIYEIIKPATANSKFPVTRLL DTRLVNQNASRWESFDVTPAVMRWTAQGHANHGFVVEVAHLEEKQGVSKRHVRISRSLHQDEHSWSQIRPLLVTFGHDGKGHPLHKREKRQAKHKQRKRLKSSC KRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLADHLNSTNHAIVQTLVNSVNSKIPKACCVPTELSAISMLYLDENEKVVLKNYQDMVVEGCGCR(SEQ ID NO:32).

[0038] In some embodiments, BMP-2 consists of the amino acids of SEQ ID NO:32.

[0039] In some embodiments of any of the chimeric polypeptides described herein, the chimeric polypeptide further comprises a linker sequence between the adjacent pair of β-TCP-binding sequences and the growth factor.

[0040] In some embodiments, the linker sequence comprises TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33).

[0041] In some embodiments, the linker sequence consists of SEQ ID NO:33.

[0042] In some embodiments, the linker sequence comprises GAGTG (SEQ ID NO: 34).

[0043] In some embodiments, the linker sequence consists of SEQ ID NO:34.

[0044] In some embodiments of any of the chimeric polypeptides described herein, the chimeric polypeptide is expressed at a K of about 1 picomolar to about 100 micromolar. DBinds to β-TCP.

[0045] Also provided herein are compositions comprising any of the chimeric polypeptides described herein.

[0046] In some embodiments, the composition further comprises β-TCP. In some embodiments, the β-TCP is formulated as a powder, putty or paste. In some embodiments, the β-TCP is placed on or in a stent, mesh or sponge.

[0047] In some embodiments of any of the compositions described herein, the composition is a pharmaceutical composition.

[0048] Also provided herein are kits comprising any of the compositions described herein.

[0049] Provided herein are chimeric polypeptides comprising: (i) one or more β-TCP-binding sequences comprising the amino acid sequence of Formula I: A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein: A0 is V, L, I, G, S, T or A; B0 is I, L, V, Q, T, S, G or A; C0 is G, A, V or S; D0 is E, D, L or G; E0 is S, T, P, T, E or D; F0 is T or S; G0 is H, T or S; H0 is H or T; I0 is R, S, K, P or H; J0 is P, S, R or K; K0 is W, F, S, P, V, A or G; and L0 is absent or is S, T, G (or A); and (ii) a mammalian growth factor, wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0050] In some embodiments, the chimeric polypeptide comprises two or more β-TCP-binding sequences of Formula I.

[0051] In some embodiments, each adjacent pair of two or more β-TCP binding sequences of Formula I are directly adjacent to each other.

[0052] In some embodiments, each adjacent pair of two or more β-TCP binding sequences of Formula I are separated by a linker sequence.

[0053] In some embodiments, the chimeric polypeptide comprises at least two different β-TCP-binding sequences of Formula I.

[0054] In some embodiments, the chimeric polypeptide comprises two or more copies of the same β-TCP-binding sequence of Formula I.

[0055] In some embodiments, the chimeric polypeptide further comprises a β-TCP-binding sequence that does not include the sequence of Formula I.

[0056] In some embodiments of any of the chimeric polypeptides described herein, the mammalian growth factor is selected from the group consisting of: EGF, PDGF, IGF-1, FGF, FGF2, FGF18, TGF-α, TGF-β, TGF-β1, TGF-β3, OP-1, OP-2, OP-3, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-9, BMP-10, BMP-12, BMP-11, BMP-13, BMP-15, DPP, Vgr, GDF -1, GDF-3, GDF-5, GDF-6, GDF-7, GDF8, GDF11, GDF15, VEGF, HABP, CBP, FGF-18, KGF, TNFα, TRAIL, WNT1, WNT2, WNT2B , WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, and WNT16.

[0057] In some embodiments, the mammalian growth factor is BMP-2. In some embodiments, BMP-2 includes MVAGTRCLLALLLPQVLLGGAAGLVPELGRRKFAAASSGRPSSQPSDEVLSEFELRLLSMFGLKQRPTPSRDAVVPPYMLDLYRRHSG QPGSPAPDHRLERAASRANTVRSFHHEESLEELPETSGKTTRRFFFNLSSIPTEEFITSAELQVFREQMQDALGNNSSFHHRINIYEIIKPATANSKFPVTRLL DTRLVNQNASRWESFDVTPAVMRWTAQGHANHGFVVEVAHLEEKQGVSKRHVRISRSLHQDEHSWSQIRPLLVTFGHDGKGHPLHKREKRQAKHKQRKRLKSSC KRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLADHLNSTNHAIVQTLVNSVNSKIPKACCVPTELSAISMLYLDENEKVVLKNYQDMVVEGCGCR(SEQ ID NO:32).

[0058] In some embodiments, BMP-2 consists of the amino acids of SEQ ID NO:32.

[0059] In some embodiments of any of the chimeric polypeptides described herein, the chimeric polypeptide further comprises a linker sequence between the adjacent pair of β-TCP-binding sequences and the growth factor.

[0060] In some embodiments, the linker sequence comprises TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33).

[0061] In some embodiments, the linker sequence consists of SEQ ID NO:33.

[0062] In some embodiments, the linker sequence comprises GSGATG (SEQ ID NO: 34).

[0063] In some embodiments, the linker sequence consists of SEQ ID NO:34.

[0064] In some embodiments of any of the chimeric polypeptides described herein, the chimeric polypeptide is expressed at a K of about 1 picomolar to about 100 micromolar. D Binds to β-TCP.

[0065] Also provided herein are compositions comprising any of the chimeric polypeptides described herein.

[0066] In some embodiments, the composition further comprises β-TCP. In some embodiments, the β-TCP is formulated as a powder, putty or paste. In some embodiments, the β-TCP is placed on or in a stent, mesh or sponge.

[0067] In some embodiments of any of the compositions described herein, the composition is a pharmaceutical composition.

[0068] Also provided herein are kits comprising any of the compositions described herein.

[0069] Provided herein are methods of promoting bone or cartilage formation in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any of the compositions described herein.

[0070] Also provided herein are methods of replacing and / or repairing bone or cartilage in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any of the compositions described herein.

[0071] Also provided herein are methods of treating bone fracture or bone loss in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any of the compositions described herein.

[0072] In some embodiments of any of the methods described herein, the subject suffers from a bone fracture. In some embodiments of any of the methods described herein, the subject suffers from a bone defect. In some embodiments of any of the methods described herein, the subject suffers from a cartilage tear or cartilage defect. In some embodiments of any of the methods described herein, the subject suffers from a cartilage loss.

[0073] Also provided herein are β-tricalcium phosphate-binding polypeptides comprising at least one sequence selected from the group consisting of VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19).

[0074] Also provided herein are β-tricalcium phosphate-binding polypeptides comprising the amino acid sequence of Formula I: A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein: A0 is V, L, I, G, S, T or A; B0 is I, L, V, Q, T, S, G or A; C0 is G, A, V or S; D0 is E, D, L or G; E0 is S, T, P, T, E or D; F0 is T or S; G0 is H, T or S; H0 is H or T; I0 is R, S, K, P or H; J0 is P, S, R or K; K0 is W, F, S, P, V, A or G; and L0 is absent or is S, T, G (or A); wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0075] As used herein, the term "subject" refers to any mammal. Therefore, the subject refers to, for example, mice, rats, dogs, cats, horses, cattle, pigs, guinea pigs, rats, humans, monkeys, etc. When the subject is a human, the subject may be referred to as a patient herein. In some embodiments, the subject or "subject in need of treatment" may be a canine (e.g., dog), a feline (e.g., cat), an equine (e.g., horse), a sheep, a cattle, a pig, a goat, a primate (e.g., a monkey (e.g., a marmoset, a baboon) or an ape (e.g., a gorilla, a chimpanzee, an orangutan or a gibbon)), a human, or a rodent (e.g., a mouse, a guinea pig, a hamster or a rat). In some embodiments, the subject or "subject in need of treatment" may be a non-human mammal, especially a mammal (e.g., mouse, rabbit (lapine), pig, dog or primate) conventionally used as a model for demonstrating therapeutic efficacy in humans.

[0076] The term "bone defect" refers to a lack or absence (e.g., partial absence) of bone at an anatomical location of a subject where bone was present in a control healthy subject. The bone defect may be the result of, for example, an infection (e.g., osteomyelitis), a tumor, trauma, or an adverse event of treatment. The bone defect may also affect the muscles, soft tissues, tendons, or joints surrounding the bone defect and cause injury. In some embodiments, the bone defect includes damage to soft tissue.

[0077] The term "cartilage defect" refers to a lack or absence (e.g., partial absence) of cartilage at an anatomical location in a subject where cartilage was present in a control healthy subject. The cartilage defect may be the result of, for example, disease, osteochondritis, osteonecrosis, or trauma. For example, a cartilage defect may affect the knee joint.

[0078] "Affinity" refers to the strength of the sum of non-covalent interactions between a β-TCP binding sequence (or a chimeric polypeptide or a polypeptide comprising a β-TCP binding sequence) and its binding partner (e.g., β-TCP). Affinity can be measured by conventional methods known in the art, including those described herein. Affinity can be measured, for example, using surface plasmon resonance (SPR) techniques (e.g., ) or biolayer interferometry (e.g. ) is determined. Additional methods for determining affinity are known in the art.

[0079] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Methods and materials for use in the present invention are described herein; other suitable methods and materials known in the art may also be used. The materials, methods, and examples are illustrative only and not restrictive. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated herein by reference in their entirety. In the event of a conflict, the present specification (including definitions) shall prevail.

[0080] Other features and advantages of the invention will be apparent from the following detailed description and drawings, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1A are representative in vivo radiographs of animals from experimental group A (mediolateral orientation).

[0082] Figure 1B are representative in vivo radiographs of animals from experimental group B (mediolateral orientation).

[0083] Figure 1C are representative in vivo radiographs of animals from experimental group C (mediolateral orientation).

[0084] Figure 2 Are representative graphs of mean radiographic scores (+SEM, n=9-10) of animals from each group at four and eight weeks post-surgery. At 4 weeks, *P<0.01 for group C1; #P<0.01 for group V; ┼P<0.01 for group C.

[0085] Figure 3A Representative radiographs, sagittal, and transverse micro-CT images of animals in Group A, B, or C at 4 weeks after surgery.

[0086] Figure 3B Representative radiographs, sagittal, and transverse micro-CT images of animals in Group A, B, or C at 4 weeks after surgery.

[0087] Figure 4 is a schematic representation of the different callus formations at 4 weeks post-surgery in animals from Group A, B or C. The red shaded area is an approximation of the mineralized callus volume.

[0088] Figure 5A are representative images of micro-CT analysis showing total callus volume in bone specimens obtained from animals in Group A (n=9), B (n=7), or C (n=8) at 8 weeks after surgery. Data are presented as mean ± SEM; *P<0.01 for B, #P<0.0.1 for C.

[0089] Figure 5B are representative images of micro-CT analysis showing bone mineral volume in bone specimens obtained from animals in Group A (n=9), B (n=7), or C (n=8) at 8 weeks after surgery. Data are presented as mean ± SEM; *P<0.01 for B, #P<0.0.1 for C.

[0090] Fig. 6A Representative images of micro-CT analysis of diaphyseal region volume of bone specimens obtained in animals of Group A (n=10), B (n=8), or C (n=7) at 4 weeks after surgery. Bone mineral volume is expressed as a percentage of bone mineral volume to total volume. Data are expressed as mean ± SEM; *P<0.05 for B, #P<0.0.5 for C. A1 vs. B1; p=0.059.

[0091] Figure 6B Representative images of micro-CT analysis of cortical bone volume of bone samples obtained in animals of Group A (n=10), B (n=8), or C (n=7) at 4 weeks after surgery. Bone mineral volume is expressed as the percentage of bone mineral volume to total volume. Data are expressed as mean ± SEM; *P<0.05 for B, #P<0.0.5 for C.

[0092] Fig. 7A Representative images of micro-CT analysis of total callus volume of bone specimens obtained in animals of Group A (n=9), B (n=7), or C (n=8) at 8 weeks after surgery. Data are presented as mean ± SEM; *P<0.01 for B, #P<0.0.1 for C.

[0093] Figure 7B Representative images of micro-CT analysis of bone mineral volume of bone specimens obtained in animals of Group A (n=9), B (n=7), or C (n=8) at 8 weeks after surgery. Data are presented as mean ± SEM; *P<0.01 for B, #P<0.0.1 for C.

[0094] Figure 8 Representative graphs of ultimate failure load (ULF) at 4 and 8 weeks post-surgery for animals in Groups A, B, or C. Data are presented as mean + / - SEM. *P < 0.05 for the 4-week group; #P < 0.01 for the 8-week group.

[0095] FIG. 9A is a representative histological image of Von Kossa-stained and H&E-stained bone specimens obtained from animals of Group A, B, or C 4 weeks after surgery.

[0096] Figure 9B is a representative histological image of TRAP+ osteoclast-stained bone specimens obtained 4 weeks after surgery from animals in Groups A, B, or C. White arrows indicate osteoclasts and yellow arrows indicate osteocytes.

[0097] 10A is a representative Von Kossa-stained and H&E-stained histological image of bone specimens obtained from animals of Groups A, B, or C at 8 weeks after surgery.

[0098] 10B is a representative TRAP+ osteoclast-stained histological image of a bone specimen obtained 8 weeks after surgery from an animal of Group A, B, or C. White arrows indicate osteoclasts, and yellow arrows indicate osteocytes.

[0099] Fig.11 is a representative immunoblot of MCF-7 cell lysates probed with anti-P-Akt antibody, where MCF-7 cells were incubated with β-TCBP-HRG.

[0100] Fig.12 Representative immunoblots of MCF-7 lysates probed with anti-P-Akt antibody where MCF-7 cells were incubated with β-TCBP-HRG (free) or β-TCBP-HRG (β-TCP bound). DETAILED DESCRIPTION

[0101] Provided herein are chimeric polypeptides comprising: (i) one or more (e.g., two, three, four, five, six, seven, eight, nine or ten) β-tricalcium phosphate (βTCP)-binding sequences, wherein at least one of the one or more β-TCP-binding sequences is selected from the group consisting of: VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), GSSGESTHHKPST (SEQ ID NO: 15); NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18), and GGADTTHHRPGT (SEQ ID NO: 19); and (ii) a mammalian growth factor (e.g., any exemplary mammalian growth factor described herein or known in the art).

[0102] Also provided herein are chimeric polypeptides comprising (i) one or more β-TCP-binding sequences comprising the amino acid sequence of Formula I: A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein:

[0103] A0 is V, L, I, G, S, T, or A;

[0104] B0 is I, L, V, Q, T, S, G, or A;

[0105] C0 is G, A, V or S;

[0106] D0 is E, D, L or G;

[0107] E0 is S, T, P, T, E, or D;

[0108] F0 is T or S;

[0109] G0 is H, T or S;

[0110] H0 is H or T;

[0111] I0 is R, S, K, P or H;

[0112] J0 is P, S, R or K;

[0113] K0 is W, F, S, P, V, A, or G; and

[0114] L0 does not exist or is S, T, G (or A); and

[0115] (ii) a mammalian growth factor (e.g., any of the exemplary mammalian growth factors described herein or known in the art),

[0116] Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0117] Also provided herein are β-tricalcium phosphate-binding polypeptides comprising at least one (e.g., two, three, four, five, six, seven, eight, nine, or ten) selected from the group consisting of VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15); NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18), and GGADTTHHRPGT (SEQ ID NO: 19). Some embodiments of these polypeptides may also include mammalian growth factors (e.g., any exemplary mammalian growth factors described herein or known in the art),

[0118] Also provided herein are β-tricalcium phosphate-binding polypeptides comprising the amino acid sequence of Formula I:

[0119] A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein:

[0120] A0 is V, L, I, G, S, T, or A;

[0121] B0 is I, L, V, Q, T, S, G, or A;

[0122] C0 is G, A, V or S;

[0123] D0 is E, D, L or G;

[0124] E0 is S, T, P, T, E, or D;

[0125] F0 is T or S;

[0126] G0 is H, T or S;

[0127] H0 is H or T;

[0128] I0 is R, S, K, P or H;

[0129] J0 is P, S, R or K;

[0130] K0 is W, F, S, P, V, A, or G; and

[0131] L0 does not exist or is S, T, G (or A);

[0132] Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1). Some embodiments of these polypeptides may also include a mammalian growth factor (e.g., any exemplary mammalian growth factor described herein or known in the art). In some embodiments, any chimeric polypeptide described herein may be bound to one or more substrates comprising βTCP (e.g., Mastergraft strips, Vitoss foam packaging, chronOS strips, vitoss Micromorsels, LifeInk500, superelastic bone, bioactive glass, βTCP powder, βTCP spray-dried powder, hydroxyapatite powder, hydroxyapatite-coated bone screws, βTCP particles, hydroxyapatite particles, and ReBOSSIS). In some embodiments, any chimeric polypeptide described herein may be bound to silicon nitride or a substrate comprising silicon nitride.

[0133] Also provided herein are compositions comprising any chimeric polypeptide or β-tricalcium phosphate-binding polypeptide described herein. In some embodiments, the composition may also include a substrate comprising βTCP (e.g., absorbable artificial bone strips, Vitoss foam packaging, chronOS strips, vitoss Micromorsels, LifeInk500, superelastic bone, bioactive glass, βTCP powder, βTCP spray-dried powder, hydroxyapatite powder, hydroxyapatite-coated bone screws, βTCP particles, hydroxyapatite particles, and ReBOSSIS). In some embodiments, the composition may also include a substrate comprising silicon nitride.

[0134] Kits comprising any of these compositions are also provided herein. Methods of promoting bone or cartilage formation in a subject in need thereof, methods of replacing and / or repairing bone or cartilage in a subject in need thereof, and methods of treating fractures or bone loss in a subject in need thereof (including administering any of the compositions described herein to the subject) are also provided.

[0135] Non-limiting aspects of these polypeptides, compositions, kits and methods are described below and may be used in any combination without limitation. Additional aspects of these polypeptides, compositions, kits and methods are known in the art.

[0136] Chimeric polypeptide

[0137] The chimeric polypeptides described herein include a β-TCP binding sequence and can bind to β-TCP (eg, any exemplary type of β-TCP described herein or known in the art). In some examples, the chimeric polypeptide may include: (i) one or more (e.g., two, three, four, five, six, seven, eight, nine, or ten) β-tricalcium phosphate (βTCP)-binding sequences, wherein at least one of the one or more β-TCP-binding sequences is selected from the group consisting of: VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), GSSGESTHHKPST (SEQ ID NO: 15); NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18), and GGADTTHHRPGT (SEQ ID NO: 19); and (ii) a mammalian growth factor (e.g., any exemplary mammalian growth factor described herein or known in the art).

[0138] In some examples, the chimeric polypeptide may include: (i) one or more β-TCP-binding sequences comprising the amino acid sequence of Formula I: A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein:

[0139] A0 is V, L, I, G, S, T, or A;

[0140] B0 is I, L, V, Q, T, S, G, or A;

[0141] C0 is G, A, V or S;

[0142] D0 is E, D, L or G;

[0143] E0 is S, T, P, T, E, or D;

[0144] F0 is T or S;

[0145] G0 is H, T or S;

[0146] H0 is H or T;

[0147] I0 is R, S, K, P or H;

[0148] J0 is P, S, R or K;

[0149] K0 is W, F, S, P, V, A, or G; and

[0150] LO is absent or is S, T, G or A; and (ii) a mammalian growth factor (eg, any exemplary mammalian growth factor described herein or known in the art), wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0151] Non-limiting examples of β-TCP binding sequences that may be present in any of the chimeric polypeptides described herein are listed in Table A below.

[0152] Table A. Exemplary β-TCP binding sequences

[0153]

[0154]

[0155]

[0156] In some embodiments, any chimeric polypeptide described herein can include two or more (e.g., three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, or twenty or more; or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22) β-TCP-binding sequences (e.g., any exemplary β-TCP-binding sequences described herein). In some embodiments, each adjacent pair of the two or more β-TCP binding sequences is directly adjacent to each other. In some embodiments, each adjacent pair of the two or more β-TCP binding sequences (e.g., any exemplary β-TCP-binding sequences described herein) is separated by a linker sequence.

[0157] In some embodiments, the chimeric polypeptide can include at least two (e.g., at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten; or 2, 3, 4, 5, 6, 7, 8, 9, or 10) different β-TCP-binding sequences (e.g., any of the exemplary β-TCP-binding sequences described herein).

[0158] In some embodiments, at least one of the two or more different β-TCP-binding sequences is LLADTTHHRPWT (SEQ ID NO: 1) or GQVLPTTTPSSP (SEQ ID NO: 19). In some embodiments, at least two different β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1) and VIGESTHHRPWS (SEQ ID NO: 2). In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWS (SEQ ID NO: 20).

[0159] In some embodiments, the at least two different β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), and IIGESSHHKPFT (SEQ ID NO: 6). In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFT (SEQ ID NO: 21).

[0160] In some embodiments, the at least two different β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), and ILAESTHHKPWT (SEQ ID NO: 4). In some embodiments, the chimeric polypeptide includes the following sequence: LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRPW GILAESTHHKPWT (SEQ ID NO: 22).

[0161] In some embodiments, the at least two β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1) and ILAESTHHKPWT (SEQ ID NO: 4). In some embodiments, the chimeric polypeptide includes at least one copy of the sequence LLADTTHHRPWTILAESTHHKPWT (SEQ ID NO: 23). In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTILAESTHHKPWTLLADTTHHRPWTILAESTHHKP WTLLADTTHHRPWT (SEQ ID NO: 24).

[0162] In some embodiments, at least two β-TCP-binding sequences include LLADTTHHRPWT (SEQ ID NO: 1) and GLGDTTHHRPWG (SEQ ID NO: 7). In some embodiments, the chimeric polypeptide includes at least one copy of the sequence of LLADTTHHRPWTGLGDTTHHRPWG (SEQ ID NO: 25). In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 26). In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 27). In some embodiments, the chimeric polypeptide includes the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 28).

[0163] In some embodiments, the at least two different β-TCP-binding sequences include STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), and GSSGESTHHKPST (SEQ ID NO: 14). In some embodiments, the chimeric polypeptide includes the sequence of STADTSHHRPSTSGGESTHHRPSTSGGESSHHKPSTGSGDSSHHRPSGSSGESTHHKPST (SEQ ID NO: 29).

[0164] In some embodiments, the at least two different β-TCP-binding sequences include VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19). In some embodiments, the chimeric polypeptide includes the sequence of VGADSTHHRPVTGAADTTHHRPVTAGADTTHHRPVTGGADTTHHR PATGGADTTHHRPGT (SEQ ID NO: 30).

[0165] In some embodiments, the at least two different β-TCP-binding sequences include STADTSHHRPS (SEQ ID NO: 10), LLADTTHHRPWT (SEQ ID NO: 1), TSGGESTHHRPS (SEQ ID NO: 11), VGADSTHHRPVT (SEQ ID NO: 15), TSGGESSHHKPS (SEQ ID NO: 12), GAADTTHHRPVT (SEQ ID NO: 16), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPS (SEQ ID NO: 14), and TGGADTTHHRPAT (SEQ ID NO: 18). In some embodiments, the chimeric polypeptide includes the following sequence: STADTSHHRPSLLADTTHHRPWTTSGGESTHHRPSVGADSTHHRPVT TSGGESSHHKPSGAADTTHHRPVTTGSGDSSHHRPSGSSGESTHHKPSSTGGADTTHHRPAT (SEQ ID NO: 31).

[0166] In some embodiments, the chimeric polypeptide can include two or more (e.g., three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, or twenty or more; or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) copies of the same β-TCP-binding sequence (e.g., any of the exemplary β-TCP-binding sequences described herein).

[0167] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:1.

[0168] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:2.

[0169] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:3.

[0170] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:4.

[0171] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:5.

[0172] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:6.

[0173] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:7.

[0174] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:8.

[0175] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:9.

[0176] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:10.

[0177] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 11.

[0178] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:12.

[0179] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:13.

[0180] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:14.

[0181] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:15.

[0182] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:16.

[0183] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:17.

[0184] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:18.

[0185] In some embodiments, the chimeric polypeptide may include a β-TCP-binding sequence having or including an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:19.

[0186] In some embodiments, the chimeric polypeptide further comprises a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) between adjacent pairs of β-TCP binding sequences (e.g., any exemplary β-TCP binding sequences described herein) and a mammalian growth factor (e.g., any exemplary mammalian growth factor described herein). In some embodiments, the linker sequence may comprise the sequence of TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33). In some embodiments, the linker sequence consists of TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33). In some embodiments, the linker sequence comprises the sequence of GSGATG (SEQ ID NO: 34). In some embodiments, the linker sequence consists of GSGATG (SEQ ID NO: 34).

[0187] In some embodiments, where the chimeric polypeptide comprises two or more β-TCP-binding sequences, a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) can be between any pair (e.g., all pairs) of adjacent β-TCP-binding sequences.

[0188] In some embodiments, the chimeric polypeptide may include a signal sequence at its N-terminus. In some embodiments, the chimeric polypeptide may also include a tag sequence (e.g., a poly-His tag, chitin binding protein (CBP), maltose binding protein (MBP), a streptococcal tag, glutathione-S-transferase (GST), thioredoxin, or an Fc region). Additional examples of tags are known in the art.

[0189] The chimeric polypeptides described herein can be present in an amount of about 1 pM to about 100 μM (e.g., about 1 pM to about 95 μM, about 1 pM to about 90 μM, about 1 pM to about 85 μM, about 1 pM to about 80 μM, about 1 pM to about 75 μM, about 1 pM to about 70 μM, about 1 pM to about 65 μM, about 1 pM to about 60 μM, about 1 pM to about 55 μM, about 1 pM to about 50 μM, about 1 pM to about 45 μM, about 1 pM to about 40 μM, about 1 pM to about 45 μM, about 1 pM to about 40 μM, about 1 pM to about 35 μM, about 1 pM to about 30 μM, about 1 pM to about 25 μM, about 1 pM to about 20 μM, about 1 pM to about 15 μM, about 1 pM to about 10 μM, about 1pM to about 5μM, about 1pM to about 2μM, about 1pM to about 1μM, about 1pM to about 950nM, about 1pM to about 900nM, about 1pM to about 850nM, about 1pM to about 800nM, about 1pM to about 750nM, about 1pM to about 700nM, about 1pM to about 650nM, about 1pM to about 600nM, about 1pM to about 550nM, about 1pM to about 500nM, about 1pM to about 450nM, about 1pM to about 400nM, about 1pM to about 350nM, about 1pM to about 300nM, about 1pM to about 250nM, about 1pM to about 200nM, about 1pM to about 150nM, about 1pM to about 100nM, about 1pM to about From about 1pM to about 95nM, from about 1pM to about 90nM, from about 1pM to about 85nM, from about 1pM to about 80nM, from about 1pM to about 75nM, from about 1pM to about 70nM, from about 1pM to about 75nM, from about 1pM to about 70nM, from about 1pM to about 65nM, from about 1pM to about 60nM, from about 1pM to about 55nM, from about 1pM to about 50nM, from about 1pM to about 45nM, from about 1pM to about 40nM, from about 1pM to about 35nM, from about 1pM to about 30nM, from about 1pM to about 25nM, from about 1pM to about 20nM, from about 1pM to about 15nM, from about 1pM to about 10nM, from about 1pM to about 5nM, from about 1pM to about 2nM, from about 1pM to about 1nM, 1pM to about 950pM, about 1pM to about 900pM, about 1pM to about 850pM, about 1pM to about 800pM, about 1pM to about 750pM, about 1pM to about 700pM, about 1pM to about 650pM, about 1pM to about 600pM, about 1pM to about 550pM, about 1pM to about 500pM, about 1pM to about 450pM, about 1pM to about 400pM, about 1pM to about 350pM, about 1pM to about 300pM, about 1pM to about 250pM, about 1pM to about 200pM, about 1pM to about 150pM, about 1pM to about 100pM, about 1pM to about 95pM, about 1pM to about 90pM, about 1pM to about 85pM,about 1pM to about 80pM, about 1pM to about 75pM, about 1pM to about 70pM, about 1pM to about 75pM, about 1pM to about 70pM, about 1pM to about 65pM, about 1pM to about 60pM, about 1pM to about 55pM, about 1pM to about 50pM, about 1pM to about 45pM, about 1pM to about 40pM, about 1pM to about 35pM, about 1pM to about 30pM, about 1pM to about 25pM, about 1pM to about 20pM, about 1pM to about 15pM, about 1pM to about 10pM, about 1pM to about 5pM, about 1pM to about 2pM, about 2pM to about 100μM, about 2pM to about 95μM, about 2pM to about 90μM, about 2pM to about 8 5μM, about 2pM to about 80μM, about 2pM to about 75μM, about 2pM to about 70μM, about 2pM to about 65μM, about 2pM to about 60μM, about 2pM to about 55μM, about 2pM to about 50μM, about 2pM to about 45μM, about 2pM to about 40μM, about 2pM to about 45μM, about 2pM to about 40μM, about 2pM to about 40μM, about 2pM to about 35μM, about 2pM to about 30μM, about 2pM to about 25μM, about 2pM to about 20μM, about 2pM to about 15μM, about 2pM to about 10μM, about 2pM to about 5μM, about 2pM to about 2μM, about 2pM to about 1μM, about 2pM to about 950nM, about 2pM to about 900nM, about 2pM about 2pM to about 850nM, about 2pM to about 800nM, about 2pM to about 750nM, about 2pM to about 700nM, about 2pM to about 650nM, about 2pM to about 600nM, about 2pM to about 550nM, about 2pM to about 500nM, about 2pM to about 450nM, about 2pM to about 400nM, about 2pM to about 350nM, about 2pM to about 300nM, about 2pM to about 250nM, about 2pM to about 200nM, about 2pM to about 150nM, about 2pM to about 100nM, about 2pM to about 95nM, about 2pM to about 90nM, about 2pM to about 85nM, about 2pM to about 80nM, about 2pM to about 75nM, about 2pM to about about 2pM to about 10nM, about 2pM to about 5nM, about 2pM to about 2nM, about 2pM to about 10nM, about 2pM to about 5nM, about 2pM to about 11nM, about 2pM to about 12nM, about 2pM to about 13nM, about 2pM to about 14nM, about 2pM to about 15nM, about 2pM to about 10nM, about 2pM to about 5nM, about 2pM to about 2nM, about 2pM to about 1 nM, about 2pM to about 950pM, about 2pM to about 900pM, about 2pM to about 850pM, about 2pM to about 800pM, about 2pM to about 750pM,From about 2pM to about 700pM, from about 2pM to about 650pM, from about 2pM to about 600pM, from about 2pM to about 550pM, from about 2pM to about 500pM, from about 2pM to about 450pM, from about 2pM to about 400pM, from about 2pM to about 350pM, from about 2pM to about 300pM, from about 2pM to about 250pM, from about 2pM to about 200pM, from about 2pM to about 150pM, from about 2pM to about 100pM, from about 2pM to about 95pM, from about 2pM to about 90pM, from about 2pM to about 85pM, from about 2pM to about 80pM, from about 2pM to about 75pM, from about 2pM to about 70pM, from about 2pM to about 75pM, from about 2pM to about 70pM, about 65pM, about 2pM to about 60pM, about 2pM to about 55pM, about 2pM to about 50pM, about 2pM to about 45pM, about 2pM to about 40pM, about 2pM to about 35pM, about 2pM to about 30pM, about 2pM to about 25pM, about 2pM to about 20pM, about 2pM to about 15pM, about 2pM to about 10pM, about 2pM to about 5pM, about 5pM to about 100μM, about 5pM to about 95μM, about 5pM to about 90μM, about 5pM to about 85μM, about 5pM to about 80μM, about 5pM to about 75μM, about 5pM to about 70μM, about 5pM to about 65μM, about 5pM to about 60μM, about 5pM to about 55μM, about 5pM to about 50μM, about 5pM to about 45μM, about 5pM to about 40μM, about 5pM to about 45μM, about 5pM to about 40μM, about 5pM to about 35μM, about 5pM to about 30μM, about 5pM to about 25μM, about 5pM to about 20μM, about 5pM to about 15μM, about 5pM to about 10μM, about 5pM to about 5μM, about 5pM to about 2μM, about 5pM to about 1μM, about 5pM to about 950nM, about 5pM to about 900nM, about 5pM to about 850nM, about 5pM to about 800nM, about 5pM to about 750nM, about 5pM to about 700nM, about 5pM to about 650nM, about 5pM to about 600nM, about 5pM to about From about 5pM to about 550nM, from about 5pM to about 500nM, from about 5pM to about 450nM, from about 5pM to about 400nM, from about 5pM to about 350nM, from about 5pM to about 300nM, from about 5pM to about 250nM, from about 5pM to about 200nM, from about 5pM to about 150nM, from about 5pM to about 100nM, from about 5pM to about 95nM, from about 5pM to about 90nM, from about 5pM to about 85nM, from about 5pM to about 80nM, from about 5pM to about 75nM, from about 5pM to about 70nM, from about 5pM to about 75nM, from about 5pM to about 70nM, from about 5pM to about 65nM, from about 5pM to about 60nM, from about 5pM to about 55nM, from about 5pM to about 50nM,about 5pM to about 45nM, about 5pM to about 40nM, about 5pM to about 35nM, about 5pM to about 30nM, about 5pM to about 25nM, about 5pM to about 20nM, about 5pM to about 15nM, about 5pM to about 10nM, about 5pM to about 5nM, about 5pM to about 2nM, about 5pM to about 1nM, about 5pM to about 950pM, about 5pM to about 900pM, about 5pM to about 850pM, about 5pM to about 800pM, about 5pM to about 750pM, about 5pM to about 700pM, about 5pM to about 650pM, about 5pM to about 600pM, about 5pM to about 550pM, about 5pM to about 500pM, about 5pM to about 450pM , about 5pM to about 400pM, about 5pM to about 350pM, about 5pM to about 300pM, about 5pM to about 250pM, about 5pM to about 200pM, about 5pM to about 150pM, about 5pM to about 100pM, about 5pM to about 95pM, about 5pM to about 90pM, about 5pM to about 85pM, about 5pM to about 80pM, about 5pM to about 75pM, about 5pM to about 70pM, about 5pM to about 75pM, about 5pM to about 70pM, about 5pM to about 65pM, about 5pM to about 60pM, about 5pM to about 55pM, about 5pM to about 50pM, about 5pM to about 45pM, about 5pM to about 40pM, about 5pM to about 35pM , about 5pM to about 30pM, about 5pM to about 25pM, about 5pM to about 20pM, about 5pM to about 15pM, about 5pM to about 10pM, about 10pM to about 100μM, about 10pM to about 95μM, about 10pM to about 90μM, about 10pM to about 85μM, about 10pM to about 80μM, about 10pM to about 75μM, about 10pM to about 70μM, about 10pM to about 65μM, about 10pM to about 60μM, about 10pM to about 55μM, about 10pM to about 50μM, about 10pM to about 45μM, about 10pM to about 40μM, about 10pM to about 45μM, about 10pM to about 40μM, about 10pM to about 35μM, about 10 pM to about 30 μM, about 10 pM to about 25 μM, about 10 pM to about 20 μM, about 10 pM to about 15 μM, about 10 pM to about 10 μM, about 10 pM to about 5 μM, about 10 pM to about 2 μM, about 10 pM to about 1 μM, about 10 pM to about 950 nM, about 10 pM to about 900 nM, about 10 pM to about 850 nM, about 10 pM to about 800 nM, about 10 pM to about 750 nM, about 10 pM to about 700 nM, about 10 pM to about 650 nM, about 10 pM to about 600 nM, about 10 pM to about 550 nM, about 10 pM to about 500 nM, about 10 pM to about 450 nM, about 10 pM to about 400 nM,about 10 pM to about 350 nM, about 10 pM to about 300 nM, about 10 pM to about 250 nM, about 10 pM to about 200 nM, about 10 pM to about 150 nM, about 10 pM to about 100 nM, about 10 pM to about 95 nM, about 10 pM to about 90 nM, about 10 pM to about 85 nM, about 10 pM to about 80 nM, about 10 pM to about 75 nM, about 10 pM to about 70 nM, about 10 pM to about 75 nM, about 10 pM to about 70 nM, about 10 pM to about 70 nM, about 10 pM to about 65 nM, about 10 pM to about 60 nM, about 10 pM to about 55 nM, about 10 pM to about 50 nM, about 10 pM to about 45 nM, about 10 pM to about 40 nM, about 1 10 pM to about 35 nM, about 10 pM to about 30 nM, about 10 pM to about 25 nM, about 10 pM to about 20 nM, about 10 pM to about 15 nM, about 10 pM to about 10 nM, about 10 pM to about 5 nM, about 10 pM to about 2 nM, about 10 pM to about 1 nM, about 10 pM to about 950 pM, about 10 pM to about 900 pM, about 10 pM to about 850 pM, about 10 pM to about 800 pM, about 10 pM to about 750 pM, about 10 pM to about 700 pM, about 10 pM to about 650 pM, about 10 pM to about 600 pM, about 10 pM to about 550 pM, about 10 pM to about 500 pM, about 10 pM to about 450 pM, about 10 pM to about 400 pM, about 10 pM to about 350 pM, about 10 pM to about 300 pM, about 10 pM to about 250 pM, about 10 pM to about 200 pM, about 10 pM to about 150 pM, about 10 pM to about 100 pM, about 10 pM to about 95 pM, about 10 pM to about 90 pM, about 10 pM to about 85 pM, about 10 pM to about 80 pM, about 10 pM to about 75 pM, about 10 pM to about 70 pM, about 10 pM to about 75 pM, about 10 pM to about 70 pM, about 10 pM to about 65 pM, about 10 pM to about 60 pM, about 10 pM to about 55 pM, about 10 pM to about 50 pM, about 10 pM to about 45 pM, about 10 pM to about M to about 40pM, about 10pM to about 35pM, about 10pM to about 30pM, about 10pM to about 25pM, about 10pM to about 20pM, about 10pM to about 15pM, about 20pM to about 100μM, about 20pM to about 95μM, about 20pM to about 90μM, about 20pM to about 85μM, about 20pM to about 80μM, about 20pM to about 75μM, about 20pM to about 70μM, about 20pM to about 65μM, about 20pM to about 60μM, about 20pM to about 55μM, about 20pM to about 50μM, about 20pM to about 45μM, about 20pM to about 40μM, about 20pM to about 45μM, about 20pM to about 40μM,about 20pM to about 35μM, about 20pM to about 30μM, about 20pM to about 25μM, about 20pM to about 20μM, about 20pM to about 15μM, about 20pM to about 10μM, about 20pM to about 5μM, about 20pM to about 2μM, about 20pM to about 1μM, about 20pM to about 950nM, about 20pM to about 900nM, about 20pM to about 850nM, about 20pM to about 800nM, about 20pM to about 750nM, about 20pM to about 700nM, about 20pM to about 650nM, about 20pM to about 600nM, about 20pM to about 550nM, about 20pM to about 500nM, about 20pM to about 450nM , about 20pM to about 400nM, about 20pM to about 350nM, about 20pM to about 300nM, about 20pM to about 250nM, about 20pM to about 200nM, about 20pM to about 150nM, about 20pM to about 100nM, about 20pM to about 95nM, about 20pM to about 90nM, about 20pM to about 85nM, about 20pM to about 80nM, about 20pM to about 75nM, about 20pM to about 70nM, about 20pM to about 75nM, about 20pM to about 70nM, about 20pM to about 70nM, about 20pM to about 65nM, about 20pM to about 60nM, about 20pM to about 55nM, about 20pM to about 50nM, about 20pM to about 45nM, From about 20 pM to about 40 nM, from about 20 pM to about 35 nM, from about 20 pM to about 30 nM, from about 20 pM to about 25 nM, from about 20 pM to about 20 nM, from about 20 pM to about 15 nM, from about 20 pM to about 10 nM, from about 20 pM to about 5 nM, from about 20 pM to about 2 nM, from about 20 pM to about 1 nM, from about 20 pM to about 950 pM, from about 20 pM to about 900 pM, from about 20 pM to about 850 pM, from about 20 pM to about 800 pM, from about 20 pM to about 750 pM, from about 20 pM to about 700 pM, from about 20 pM to about 650 pM, from about 20 pM to about 600 pM, from about 20 pM to about 550 pM, from about 20 pM to about 500 pM, From about 20 pM to about 450 pM, from about 20 pM to about 400 pM, from about 20 pM to about 350 pM, from about 20 pM to about 300 pM, from about 20 pM to about 250 pM, from about 20 pM to about 200 pM, from about 20 pM to about 150 pM, from about 20 pM to about 100 pM, from about 20 pM to about 95 pM, from about 20 pM to about 90 pM, from about 20 pM to about 85 pM, from about 20 pM to about 80 pM, from about 20 pM to about 75 pM, from about 20 pM to about 70 pM, from about 20 pM to about 75 pM, from about 20 pM to about 70 pM, from about 20 pM to about 65 pM, from about 20 pM to about 60 pM, from about 20 pM to about 55 pM, from about 20 pM to about 50 pM,about 20 pM to about 45 pM, about 20 pM to about 40 pM, about 20 pM to about 35 pM, about 20 pM to about 30 pM, about 20 pM to about 25 pM, about 50 pM to about 100 μM, about 50 pM to about 95 μM, about 50 pM to about 90 μM, about 50 pM to about 85 μM, about 50 pM to about 80 μM, about 50 pM to about 75 μM, about 50 pM to about 70 μM, about 50 pM to about 65 μM, about 50 pM to about 60 μM, about 50 pM to about 55 μM, about 50 pM to about 50 μM, about 50 pM to about 45 μM, about 50 pM to about 40 μM, about 50 pM to about 3 5 μM, about 50 pM to about 30 μM, about 50 pM to about 25 μM, about 50 pM to about 20 μM, about 50 pM to about 15 μM, about 50 pM to about 10 μM, about 50 pM to about 5 μM, about 50 pM to about 2 μM, about 50 pM to about 1 μM, about 50 pM to about 950 nM, about 50 pM to about 900 nM, about 50 pM to about 850 nM, about 50 pM to about 800 nM, about 50 pM to about 750 nM, about 50 pM to about 700 nM, about 50 pM to about 650 nM, about 50 pM to about 600 nM, about 50 pM to about 550 nM, about 50 pM to about 500 nM, about 50 pM to about 450 nM, about 50 pM to about 40 0 nM, about 50 pM to about 350 nM, about 50 pM to about 300 nM, about 50 pM to about 250 nM, about 50 pM to about 200 nM, about 50 pM to about 150 nM, about 50 pM to about 100 nM, about 50 pM to about 95 nM, about 50 pM to about 90 nM, about 50 pM to about 85 nM, about 50 pM to about 80 nM, about 50 pM to about 75 nM, about 50 pM to about 70 nM, about 50 pM to about 75 nM, about 50 pM to about 70 nM, about 50 pM to about 70 nM, about 50 pM to about 65 nM, about 50 pM to about 60 nM, about 50 pM to about 55 nM, about 50 pM to about 50 nM, about 50 pM to about 45 nM, about 50 pM to about 40 nM , about 50 pM to about 35 nM, about 50 pM to about 30 nM, about 50 pM to about 25 nM, about 50 pM to about 20 nM, about 50 pM to about 15 nM, about 50 pM to about 10 nM, about 50 pM to about 5 nM, about 50 pM to about 2 nM, about 50 pM to about 1 nM, about 50 pM to about 950 pM, about 50 pM to about 900 pM, about 50 pM to about 850 pM, about 50 pM to about 800 pM, about 50 pM to about 750 pM, about 50 pM to about 700 pM, about 50 pM to about 650 pM, about 50 pM to about 600 pM, about 50 pM to about 550 pM, about 50 pM to about 500 pM, about 50 pM to about 450 pM,From about 50 pM to about 400 pM, from about 50 pM to about 350 pM, from about 50 pM to about 300 pM, from about 50 pM to about 250 pM, from about 50 pM to about 200 pM, from about 50 pM to about 150 pM, from about 50 pM to about 100 pM, from about 50 pM to about 95 pM, from about 50 pM to about 90 pM, from about 50 pM to about 85 pM, from about 50 pM to about 80 pM, from about 50 pM to about 75 pM, from about 50 pM to about 70 pM, from about 50 pM to about 75 pM, from about 50 pM to about 70 pM, from about 50 pM to about 65 pM, from about 50 pM to about 60 pM, from about 50 pM to about 55 pM, from about 100 pM to about 100 μM, from about 100 pM to about 95 μM M, about 100 pM to about 90 μM, about 100 pM to about 85 μM, about 100 pM to about 80 μM, about 100 pM to about 75 μM, about 100 pM to about 70 μM, about 100 pM to about 65 μM, about 100 pM to about 60 μM, about 100 pM to about 55 μM, about 100 pM to about 50 μM, about 100 pM to about 45 μM, about 100 pM to about 40 μM, about 100 pM to about 45 μM, about 100 pM to about 40 μM, about 100 pM to about 35 μM, about 100 pM to about 30 μM, about 100 pM to about 25 μM, about 100 pM to about 20 μM, about 100 pM to about 15 μM, about 100 pM to about 10 μM , about 100 pM to about 5 μM, about 100 pM to about 2 μM, about 100 pM to about 1 μM, about 100 pM to about 950 nM, about 100 pM to about 900 nM, about 100 pM to about 850 nM, about 100 pM to about 800 nM, about 100 pM to about 750 nM, about 100 pM to about 700 nM, about 100 pM to about 650 nM, about 100 pM to about 600 nM, about 100 pM to about 550 nM, about 100 pM to about 500 nM, about 100 pM to about 450 nM, about 100 pM to about 400 nM, about 100 pM to about 350 nM, about 100 pM to about 300 nM, about 100 pM to about 250 nM, about 100 pM to about 200 nM, about 100 pM to about 150 nM, about 100 pM to about 100 nM, about 100 pM to about 95 nM, about 100 pM to about 90 nM, about 100 pM to about 85 nM, about 100 pM to about 80 nM, about 100 pM to about 75 nM, about 100 pM to about 70 nM, about 100 pM to about 75 nM, about 100 pM to about 70 nM, about 100 pM to about 70 nM, about 100 pM to about 65 nM, about 100 pM to about 60 nM, about 100 pM to about 55 nM, about 100 pM to about 50 nM, about 100 pM to about 45 nM, about 100 pM to about 40 nM, about 100 pM to about 35 nM, about 100 pM to about 30 nM,From about 100 pM to about 25 nM, from about 100 pM to about 20 nM, from about 100 pM to about 15 nM, from about 100 pM to about 10 nM, from about 100 pM to about 5 nM, from about 100 pM to about 2 nM, from about 100 pM to about 1 nM, from about 100 pM to about 950 pM, from about 100 pM to about 900 pM, from about 100 pM to about 850 pM, from about 100 pM to about 800 pM, from about 100 pM to about 750 pM, from about 100 pM to about 700 pM, from about 100 pM to about 650 pM, from about 100 pM to about 600 pM, from about 100 pM to about 550 pM, from about 100 pM to about 500 pM, from about 100 pM to about 450 pM, from about 100 pM to about From about 100 pM to about 400 pM, from about 100 pM to about 350 pM, from about 100 pM to about 300 pM, from about 100 pM to about 250 pM, from about 100 pM to about 200 pM, from about 100 pM to about 150 pM, from about 200 pM to about 100 μM, from about 200 pM to about 95 μM, from about 200 pM to about 90 μM, from about 200 pM to about 85 μM, from about 200 pM to about 80 μM, from about 200 pM to about 75 μM, from about 200 pM to about 70 μM, from about 200 pM to about 65 μM, from about 200 pM to about 60 μM, from about 200 pM to about 55 μM, from about 200 pM to about 50 μM, from about 200 pM to about 45 μM, from about 200 pM to about 40 μM, From about 200 pM to about 45 μM, from about 200 pM to about 40 μM, from about 200 pM to about 35 μM, from about 200 pM to about 30 μM, from about 200 pM to about 25 μM, from about 200 pM to about 20 μM, from about 200 pM to about 15 μM, from about 200 pM to about 10 μM, from about 200 pM to about 5 μM, from about 200 pM to about 2 μM, from about 200 pM to about 1 μM, from about 200 pM to about 950 nM, from about 200 pM to about 900 nM, from about 200 pM to about 850 nM, from about 200 pM to about 800 nM, from about 200 pM to about 750 nM, from about 200 pM to about 700 nM, from about 200 pM to about 650 nM, from about 200 pM to about 600 nM 0 nM, about 200 pM to about 550 nM, about 200 pM to about 500 nM, about 200 pM to about 450 nM, about 200 pM to about 400 nM, about 200 pM to about 350 nM, about 200 pM to about 300 nM, about 200 pM to about 250 nM, about 200 pM to about 200 nM, about 200 pM to about 150 nM, about 200 pM to about 100 nM, about 200 pM to about 95 nM, about 200 pM to about 90 nM, about 200 pM to about 85 nM, about 200 pM to about 80 nM, about 200 pM to about 75 nM, about 200 pM to about 70 nM, about 200 pM to about 75 nM, about 200 pM to about 70 nM,about 200 pM to about 65 nM, about 200 pM to about 60 nM, about 200 pM to about 55 nM, about 200 pM to about 50 nM, about 200 pM to about 45 nM, about 200 pM to about 40 nM, about 200 pM to about 35 nM, about 200 pM to about 30 nM, about 200 pM to about 25 nM, about 200 pM to about 20 nM, about 200 pM to about 15 nM, about 200 pM to about 10 nM, about 200 pM to about 5 nM, about 200 pM to about 2 nM, about 200 pM to about 1 nM, about 200 pM to about 950 pM, about 200 pM to about 900 pM, about 200 pM to about 850 pM, about 200 pM to about 800 pM , about 200 pM to about 750 pM, about 200 pM to about 700 pM, about 200 pM to about 650 pM, about 200 pM to about 600 pM, about 200 pM to about 550 pM, about 200 pM to about 500 pM, about 200 pM to about 450 pM, about 200 pM to about 400 pM, about 200 pM to about 350 pM, about 200 pM to about 300 pM, about 200 pM to about 250 pM, about 300 pM to about 100 μM, about 300 pM to about 95 μM, about 300 pM to about 90 μM, about 300 pM to about 85 μM, about 300 pM to about 80 μM, about 300 pM to about 75 μM, about 300 pM to about 70 μM, about 0 μM, about 300 pM to about 65 μM, about 300 pM to about 60 μM, about 300 pM to about 55 μM, about 300 pM to about 50 μM, about 300 pM to about 45 μM, about 300 pM to about 40 μM, about 300 pM to about 45 μM, about 300 pM to about 40 μM, about 300 pM to about 35 μM, about 300 pM to about 30 μM, about 300 pM to about 25 μM, about 300 pM to about 20 μM, about 300 pM to about 15 μM, about 300 pM to about 10 μM, about 300 pM to about 5 μM, about 300 pM to about 2 μM, about 300 pM to about 1 μM, about 300 pM to about 950 nM, about 300 pM to about 900 nM, about 3 From about 300 pM to about 850 nM, from about 300 pM to about 800 nM, from about 300 pM to about 750 nM, from about 300 pM to about 700 nM, from about 300 pM to about 650 nM, from about 300 pM to about 600 nM, from about 300 pM to about 550 nM, from about 300 pM to about 500 nM, from about 300 pM to about 450 nM, from about 300 pM to about 400 nM, from about 300 pM to about 350 nM, from about 300 pM to about 300 nM, from about 300 pM to about 250 nM, from about 300 pM to about 200 nM, from about 300 pM to about 150 nM, from about 300 pM to about 100 nM, from about 300 pM to about 95 nM, from about 300 pM to about 90 nM,about 300 pM to about 85 nM, about 300 pM to about 80 nM, about 300 pM to about 75 nM, about 300 pM to about 70 nM, about 300 pM to about 75 nM, about 300 pM to about 70 nM, about 300 pM to about 65 nM, about 300 pM to about 60 nM, about 300 pM to about 55 nM, about 300 pM to about 50 nM, about 300 pM to about 45 nM, about 300 pM to about 40 nM, about 300 pM to about 35 nM, about 300 pM to about 30 nM, about 300 pM to about 25 nM, about 300 pM to about 20 nM, about 300 pM to about 15 nM, about 300 pM to about 10 nM, about 300 pM to about 5 nM, about 300 pM to about pM to about 2nM, about 300pM to about 1nM, about 300pM to about 950pM, about 300pM to about 900pM, about 300pM to about 850pM, about 300pM to about 800pM, about 300pM to about 750pM, about 300pM to about 700pM, about 300pM to about 650pM, about 300pM to about 600pM, about 300pM to about 550pM, about 300pM to about 500pM, about 300pM to about 450pM, about 300pM to about 400pM, about 300pM to about 350pM, about 400pM to about 100μM, about 400pM to about 95μM, about 400pM to about 90μM, about 400pM to about 85 μM, about 400 pM to about 80 μM, about 400 pM to about 75 μM, about 400 pM to about 70 μM, about 400 pM to about 65 μM, about 400 pM to about 60 μM, about 400 pM to about 55 μM, about 400 pM to about 50 μM, about 400 pM to about 45 μM, about 400 pM to about 40 μM, about 400 pM to about 45 μM, about 400 pM to about 40 μM, about 400 pM to about 40 μM, about 400 pM to about 35 μM, about 400 pM to about 30 μM, about 400 pM to about 25 μM, about 400 pM to about 20 μM, about 400 pM to about 15 μM, about 400 pM to about 10 μM, about 400 pM to about 5 μM, about 400 pM to about 2 μM, about 400pM to about 1 μM, about 400pM to about 950nM, about 400pM to about 900nM, about 400pM to about 850nM, about 400pM to about 800nM, about 400pM to about 750nM, about 400pM to about 700nM, about 400pM to about 650nM, about 400pM to about 600nM, about 400pM to about 550nM, about 400pM to about 500nM, about 400pM to about 450nM, about 400pM to about 400nM, about 400pM to about 350nM, about 400pM to about 300nM, about 400pM to about 250nM, about 400pM to about 200nM, about 400pM to about 150nM,about 400 pM to about 100 nM, about 400 pM to about 95 nM, about 400 pM to about 90 nM, about 400 pM to about 85 nM, about 400 pM to about 80 nM, about 400 pM to about 75 nM, about 400 pM to about 70 nM, about 400 pM to about 75 nM, about 400 pM to about 70 nM, about 400 pM to about 65 nM, about 400 pM to about 60 nM, about 400 pM to about 55 nM, about 400 pM to about 50 nM, about 400 pM to about 45 nM, about 400 pM to about 40 nM, about 400 pM to about 35 nM, about 400 pM to about 30 nM, about 400 pM to about 25 nM, about 400 pM to about 20 nM, about pM to about 800 pM, about 400 pM to about 950 pM, about 400 pM to about 900 pM, about 400 pM to about 850 pM, about 400 pM to about 800 pM, about 400 pM to about 750 pM, about 400 pM to about 700 pM, about 400 pM to about 650 pM, about 400 pM to about 600 pM, about 400 pM to about 550 pM, about 400 pM to about 500 pM, about 400 pM to about 450 pM, about 500 pM to about 100 μM, about 500 pM to about 95 μM, about 500 pM to about 90 μM, about 500 pM to about 85 μM, about 500 pM to about 80 μM, about 500 pM to about 75 μM, about 500 pM to about 70 μM, about 500 pM to about 65 μM, about 500 pM to about 60 μM, about 500 pM to about 55 μM, about 500 pM to about 50 μM, about 500 pM to about 45 μM, about 500 pM to about 40 μM, about 500 pM to about 45 μM, about 500 pM to about 40 μM, about 500 pM to about 40 μM, about 500 pM to about 35 μM, about 500 pM to about 30 μM, about 500 pM to about 25 μM, about 500 pM to about 20 μM, about 500 pM to about 15 μM, about 500 pM to about 10 μM, about 500 pM to about 5 μM. about 500 pM to about 2 μM, about 500 pM to about 1 μM, about 500 pM to about 950 nM, about 500 pM to about 900 nM, about 500 pM to about 850 nM, about 500 pM to about 800 nM, about 500 pM to about 750 nM, about 500 pM to about 700 nM, about 500 pM to about 650 nM, about 500 pM to about 600 nM, about 500 pM to about 550 nM, about 500 pM to about 500 nM, about 500 pM to about 450 nM, about 500 pM to about 400 nM, about 500 pM to about 350 nM, about 500 pM to about 300 nM, about 500 pM to about 250 nM, about 500 pM to about 200 nM,about 500 pM to about 150 nM, about 500 pM to about 100 nM, about 500 pM to about 95 nM, about 500 pM to about 90 nM, about 500 pM to about 85 nM, about 500 pM to about 80 nM, about 500 pM to about 75 nM, about 500 pM to about 70 nM, about 500 pM to about 75 nM, about 500 pM to about 70 nM, about 500 pM to about 70 nM, about 500 pM to about 65 nM, about 500 pM to about 60 nM, about 500 pM to about 55 nM, about 500 pM to about 50 nM, about 500 pM to about 45 nM, about 500 pM to about 40 nM, about 500 pM to about 35 nM, about 500 pM to about 30 nM, about 500 pM to about 25 nM, From about 500 pM to about 20 nM, from about 500 pM to about 15 nM, from about 500 pM to about 10 nM, from about 500 pM to about 5 nM, from about 500 pM to about 2 nM, from about 500 pM to about 1 nM, from about 500 pM to about 950 pM, from about 500 pM to about 900 pM, from about 500 pM to about 850 pM, from about 500 pM to about 800 pM, from about 500 pM to about 750 pM, from about 500 pM to about 700 pM, from about 500 pM to about 650 pM, from about 500 pM to about 600 pM, from about 500 pM to about 550 pM, from about 600 pM to about 100 μM, from about 600 pM to about 95 μM, from about 600 pM to about 90 μM, from about 600 pM to about 850 pM 5μM, about 600pM to about 80μM, about 600pM to about 75μM, about 600pM to about 70μM, about 600pM to about 65μM, about 600pM to about 60μM, about 600pM to about 55μM, about 600pM to about 50μM, about 600pM to about 45μM, about 600pM to about 40μM, about 600pM to about 45μM, about 600pM to about 40μM, about 600pM to about 40μM, about 600pM to about 35μM, about 600pM to about 30μM, about 600pM to about 25μM, about 600pM to about 20μM, about 600pM to about 15μM, about 600pM to about 10μM, about 600pM to about 5μM, about 600pM to about 2μM, about 600pM to about 1 μM, about 600pM to about 950nM, about 600pM to about 900nM, about 600pM to about 850nM, about 600pM to about 800nM, about 600pM to about 750nM, about 600pM to about 700nM, about 600pM to about 650nM, about 600pM to about 600nM, about 600pM to about 550nM, about 600pM to about 500nM, about 600pM to about 450nM, about 600pM to about 400nM, about 600pM to about 350nM, about 600pM to about 300nM, about 600pM to about 250nM, about 600pM to about 200nM, about 600pM to about 150nM,about 600 pM to about 100 nM, about 600 pM to about 95 nM, about 600 pM to about 90 nM, about 600 pM to about 85 nM, about 600 pM to about 80 nM, about 600 pM to about 75 nM, about 600 pM to about 70 nM, about 600 pM to about 75 nM, about 600 pM to about 70 nM, about 600 pM to about 65 nM, about 600 pM to about 60 nM, about 600 pM to about 55 nM, about 600 pM to about 50 nM, about 600 pM to about 45 nM, about 600 pM to about 40 nM, about 600 pM to about 35 nM, about 600 pM to about 30 nM, about 600 pM to about 25 nM, about 600 pM to about 20 nM, about In some embodiments, the present invention relates to a polymerizable compound of the present invention, wherein the polymerizable compound is a polymerizable compound of the present invention and / or the polymerizable compound is a polymerizable compound of the present invention. The polymerizable compound is a polymerizable compound of the present invention and / or the polymerizable compound is a polymerizable compound of the present invention. , about 700 pM to about 65 μM, about 700 pM to about 60 μM, about 700 pM to about 55 μM, about 700 pM to about 50 μM, about 700 pM to about 45 μM, about 700 pM to about 40 μM, about 700 pM to about 45 μM, about 700 pM to about 40 μM, about 700 pM to about 35 μM, about 700 pM to about 30 μM, about 700 pM to about 25 μM, about 700 pM to about 20 μM, about 700 pM to about 15 μM, about 700 pM to about 10 μM, about 700 pM to about 5 μM, about 700 pM to about 2 μM, about 700 pM to about 1 μM, about 700 pM to about 950 nM, about 700 pM to about 900 nM, about 700 pM to about 850 nM, about 700 pM to about 800 nM, about 700 pM to about 750 nM, about 700 pM to about 700 nM, about 700 pM to about 650 nM, about 700 pM to about 600 nM, about 700 pM to about 550 nM, about 700 pM to about 500 nM, about 700 pM to about 450 nM, about 700 pM to about 400 nM, about 700 pM to about 350 nM, about 700 pM to about 300 nM, about 700 pM to about 250 nM, about 700 pM to about 200 nM, about 700 pM to about 150 nM, about 700 pM to about 100 nM, about 700 pM to about 95 nM, about 700 pM to about 90 nM,about 700 pM to about 85 nM, about 700 pM to about 80 nM, about 700 pM to about 75 nM, about 700 pM to about 70 nM, about 700 pM to about 75 nM, about 700 pM to about 70 nM, about 700 pM to about 70 nM, about 700 pM to about 65 nM, about 700 pM to about 60 nM, about 700 pM to about 55 nM, about 700 pM to about 50 nM, about 700 pM to about 45 nM, about 700 pM to about 40 nM, about 700 pM to about 35 nM, about 700 pM to about 30 nM, about 700 pM to about 25 nM, about 700 pM to about 20 nM, about 700 pM to about 15 nM, about 700 pM to about 10 nM, about 700 pM to about 5 nM, about 700 pM to about 700 pM to about 100 μM, about 700 pM to about 2 nM, about 700 pM to about 1 nM, about 700 pM to about 950 pM, about 700 pM to about 900 pM, about 700 pM to about 850 pM, about 700 pM to about 800 pM, about 700 pM to about 750 pM, about 800 pM to about 100 μM, about 800 pM to about 95 μM, about 800 pM to about 90 μM, about 800 pM to about 85 μM, about 800 pM to about 80 μM, about 800 pM to about 75 μM, about 800 pM to about 70 μM, about 800 pM to about 65 μM, about 800 pM to about 60 μM, about 800 pM to about 55 μM, about 800 pM to about 50 μM, about 800 pM to about 45 μM , about 800 pM to about 40 μM, about 800 pM to about 45 μM, about 800 pM to about 40 μM, about 800 pM to about 35 μM, about 800 pM to about 30 μM, about 800 pM to about 25 μM, about 800 pM to about 20 μM, about 800 pM to about 15 μM, about 800 pM to about 10 μM, about 800 pM to about 5 μM, about 800 pM to about 2 μM, about 800 pM to about 1 μM, about 800 pM to about 950 nM, about 800 pM to about 900 nM, about 800 pM to about 850 nM, about 800 pM to about 800 nM, about 800 pM to about 750 nM, about 800 pM to about 700 nM, about 800 pM to about 650 nM. 0nM, about 800pM to about 600nM, about 800pM to about 550nM, about 800pM to about 500nM, about 800pM to about 450nM, about 800pM to about 400nM, about 800pM to about 350nM, about 800pM to about 300nM, about 800pM to about 250nM, about 800pM to about 200nM, about 800pM to about 150nM, about 800pM to about 100nM, about 800pM to about 95nM, about 800pM to about 90nM, about 800pM to about 85nM, about 800pM to about 80nM, about 800pM to about 75nM, about 800pM to about 70nM, about 800pM to about 75nM,about 800 pM to about 70 nM, about 800 pM to about 65 nM, about 800 pM to about 60 nM, about 800 pM to about 55 nM, about 800 pM to about 50 nM, about 800 pM to about 45 nM, about 800 pM to about 40 nM, about 800 pM to about 35 nM, about 800 pM to about 30 nM, about 800 pM to about 25 nM, about 800 pM to about 20 nM, about 800 pM to about 15 nM, about 800 pM to about 10 nM, about 800 pM to about 5 nM, about 800 pM to about 2 nM, about 800 pM to about 1 nM, about 800 pM to about 950 pM, about 800 pM to about 900 pM, about 800 pM to about 850 pM, about 900pM to about 100μM, about 900pM to about 95μM, about 900pM to about 90μM, about 900pM to about 85μM, about 900pM to about 80μM, about 900pM to about 75μM, about 900pM to about 70μM, about 900pM to about 65μM, about 900pM to about 60μM, about 900pM to about 55μM, about 900pM to about 50μM, about 900pM to about 45μM, about 900pM to about 40μM, about 900pM to about 45μM, about 900pM to about 40μM, about 900pM to about 40μM, about 900pM to about 35μM, about 900pM to about 30μM, about 900pM to about 25μM, about 900pM to about 20μM, about 900 pM to about 15 μM, about 900 pM to about 10 μM, about 900 pM to about 5 μM, about 900 pM to about 2 μM, about 900 pM to about 1 μM, about 900 pM to about 950 nM, about 900 pM to about 900 nM, about 900 pM to about 850 nM, about 900 pM to about 800 nM, about 900 pM to about 750 nM, about 900 pM to about 700 nM, about 900 pM to about 650 nM, about 900 pM to about 600 nM, about 900 pM to about 550 nM, about 900 pM to about 500 nM, about 900 pM to about 450 nM, about 900 pM to about 400 nM, about 900 pM to about 350 nM, about 900 pM to about 300nM, about 900pM to about 250nM, about 900pM to about 200nM, about 900pM to about 150nM, about 900pM to about 100nM, about 900pM to about 95nM, about 900pM to about 90nM, about 900pM to about 85nM, about 900pM to about 80nM, about 900pM to about 75nM, about 900pM to about 70nM, about 900pM to about 75nM, about 900pM to about 70nM, about 900pM to about 70nM, about 900pM to about 65nM, about 900pM to about 60nM, about 900pM to about 55nM, about 900pM to about 50nM, about 900pM to about 45nM, about 900pM to about 40nM,about 900 pM to about 35 nM, about 900 pM to about 30 nM, about 900 pM to about 25 nM, about 900 pM to about 20 nM, about 900 pM to about 15 nM, about 900 pM to about 10 nM, about 900 pM to about 5 nM, about 900 pM to about 2 nM, about 900 pM to about 1 nM, about 900 pM to about 950 pM, about 1 nM to about 100 μM, about 1 nM to about 95 μM, about 1 nM to about 90 μM, about 1 nM to about 85 μM, about 1 nM to about 80 μM, about 1 nM to about 75 μM, about 1 nM to about 70 μM, about 1 nM to about 65 μM, about 1 nM to about 60 μM, about 1 nM to about 55 μM, about 1 nM to about 50 μM, about 1 nM to about 45 μM, about 1 nM to about 40 μM, about 1 nM to about 45 μM, about 1 nM to about 40 μM, about 1 nM to about 35 μM, about 1 nM to about 30 μM, about 1 nM to about 25 μM, about 1 nM to about 20 μM, about 1 nM to about 15 μM, about 1 nM to about 10 μM, about 1 nM to about 5 μM, about 1 nM to about 2 μM, about 1 nM to about 1 μM, about 1 nM to about 950 nM, about 1 nM to about 900 nM, about 1 nM to about 850 nM, about 1 nM to about 800 nM, about 1 nM to about 750 nM, about 1 nM to about 700 nM, about 1 nM to about 650 nM, about 1 nM to about 600 nM, about 1 nM to about 550 nM, about 1 nM to about 500nM, about 1nM to about 450nM, about 1nM to about 400nM, about 1nM to about 350nM, about 1nM to about 300nM, about 1nM to about 250nM, about 1nM to about 200nM, about 1nM to about 150nM, about 1nM to about 100nM, about 1nM to about 95nM, about 1nM to about 90nM, about 1nM to about 85nM, about 1nM to about 80nM, about 1nM to about 75nM, about 1nM to about 70nM, about 1nM to about 75nM, about 1nM to about 70nM, about 1nM to about 65nM, about 1nM to about 60nM, about 1nM to about 55nM, about 1nM to about 50nM, about 1nM to about 45nM, about 1 nM to about 40 nM, about 1 nM to about 35 nM, about 1 nM to about 30 nM, about 1 nM to about 25 nM, about 1 nM to about 20 nM, about 1 nM to about 15 nM, about 1 nM to about 10 nM, about 1 nM to about 5 nM, about 1 nM to about 2 nM, about 2 nM to about 100 μM, about 2 nM to about 95 μM, about 2 nM to about 90 μM, about 2 nM to about 85 μM, about 2 nM to about 80 μM, about 2 nM to about 75 μM, about 2 nM to about 70 μM, about 2 nM to about 65 μM, about 2 nM to about 60 μM, about 2 nM to about 55 μM, about 2 nM to about 50 μM, about 2 nM to about 45 μM, about 2 nM to about 40 μM, about 2 nM to about 45 μM,about 2nM to about 40μM, about 2nM to about 35μM, about 2nM to about 30μM, about 2nM to about 25μM, about 2nM to about 20μM, about 2nM to about 15μM, about 2nM to about 10μM, about 2nM to about 5μM, about 2nM to about 2μM, about 2nM to about 1μM, about 2nM to about 950nM, about 2nM to about 900nM, about 2nM to about 850nM, about 2nM to about 800nM, about 2nM to about 750nM, about 2nM to about 700nM, about 2nM to about 650nM, about 2nM to about 600nM, about 2nM to about 550nM, about 2nM to about 500nM, about 2nM to about 450nM, about 2nM to about 400nM nM, about 2nM to about 350nM, about 2nM to about 300nM, about 2nM to about 250nM, about 2nM to about 200nM, about 2nM to about 150nM, about 2nM to about 100nM, about 2nM to about 95nM, about 2nM to about 90nM, about 2nM to about 85nM, about 2nM to about 80nM, about 2nM to about 75nM, about 2nM to about 70nM, about 2nM to about 75nM, about 2nM to about 70nM, about 2nM to about 70nM, about 2nM to about 65nM, about 2nM to about 60nM, about 2nM to about 55nM, about 2nM to about 50nM, about 2nM to about 45nM, about 2nM to about 40nM, about 2nM to about 35nM, about 2nM to about 30nM M, about 2nM to about 25nM, about 2nM to about 20nM, about 2nM to about 15nM, about 2nM to about 10nM, about 2nM to about 5nM, about 5nM to about 100μM, about 5nM to about 95μM, about 5nM to about 90μM, about 5nM to about 85μM, about 5nM to about 80μM, about 5nM to about 75μM, about 5nM to about 70μM, about 5nM to about 65μM, about 5nM to about 60μM, about 5nM to about 55μM, about 5nM to about 50μM, about 5nM to about 45μM, about 5nM to about 40μM, about 5nM to about 45μM, about 5nM to about 40μM, about 5nM to about 35μM, about 5nM to about 30μM, about 5nM to about 25 μM, about 5 nM to about 20 μM, about 5 nM to about 15 μM, about 5 nM to about 10 μM, about 5 nM to about 5 μM, about 5 nM to about 2 μM, about 5 nM to about 1 μM, about 5 nM to about 950 nM, about 5 nM to about 900 nM, about 5 nM to about 850 nM, about 5 nM to about 800 nM, about 5 nM to about 750 nM, about 5 nM to about 700 nM, about 5 nM to about 650 nM, about 5 nM to about 600 nM, about 5 nM to about 550 nM, about 5 nM to about 500 nM, about 5 nM to about 450 nM, about 5 nM to about 400 nM, about 5 nM to about 350 nM, about 5 nM to about 300 nM, about 5 nM to about 250 nM,about 5nM to about 200nM, about 5nM to about 150nM, about 5nM to about 100nM, about 5nM to about 95nM, about 5nM to about 90nM, about 5nM to about 85nM, about 5nM to about 80nM, about 5nM to about 75nM, about 5nM to about 70nM, about 5nM to about 75nM, about 5nM to about 70nM, about 5nM to about 70nM, about 5nM to about 65nM, about 5nM to about 60nM, about 5nM to about 55nM, about 5nM to about 50nM, about 5nM to about 45nM, about 5nM to about 40nM, about 5nM to about 35nM, about 5nM to about 30nM, about 5nM to about 25nM, about 5nM to about 20nM, about 5nM to about 15nM, about 5nM to about M to about 10 nM, about 10 nM to about 100 μM, about 10 nM to about 95 μM, about 10 nM to about 90 μM, about 10 nM to about 85 μM, about 10 nM to about 80 μM, about 10 nM to about 75 μM, about 10 nM to about 70 μM, about 10 nM to about 65 μM, about 10 nM to about 60 μM, about 10 nM to about 55 μM, about 10 nM to about 50 μM, about 10 nM to about 45 μM, about 10 nM to about 40 μM, about 10 nM to about 45 μM, about 10 nM to about 40 μM, about 10 nM to about 35 μM, about 10 nM to about 30 μM, about 10 nM to about 25 μM, about 10 nM to about 20 μM, about 10 nM to about 15 μM , about 10 nM to about 10 μM, about 10 nM to about 5 μM, about 10 nM to about 2 μM, about 10 nM to about 1 μM, about 10 nM to about 950 nM, about 10 nM to about 900 nM, about 10 nM to about 850 nM, about 10 nM to about 800 nM, about 10 nM to about 750 nM, about 10 nM to about 700 nM, about 10 nM to about 650 nM, about 10 nM to about 600 nM, about 10 nM to about 550 nM, about 10 nM to about 500 nM, about 10 nM to about 450 nM, about 10 nM to about 400 nM, about 10 nM to about 350 nM, about 10 nM to about 300 nM, about 10 nM to about 250 nM, about 10 nM to about 10 nM, about 10 nM to about 150 nM, about 10 nM to about 100 nM, about 10 nM to about 95 nM, about 10 nM to about 90 nM, about 10 nM to about 85 nM, about 10 nM to about 80 nM, about 10 nM to about 75 nM, about 10 nM to about 70 nM, about 10 nM to about 75 nM, about 10 nM to about 70 nM, about 10 nM to about 65 nM, about 10 nM to about 60 nM, about 10 nM to about 55 nM, about 10 nM to about 50 nM, about 10 nM to about 45 nM, about 10 nM to about 40 nM, about 10 nM to about 35 nM, about 10 nM to about 30 nM, about 10 nM to about 25 nM, about 10 nM to about 20 nM,about 10 nM to about 15 nM, about 20 nM to about 100 μM, about 20 nM to about 95 μM, about 20 nM to about 90 μM, about 20 nM to about 85 μM, about 20 nM to about 80 μM, about 20 nM to about 75 μM, about 20 nM to about 70 μM, about 20 nM to about 65 μM, about 20 nM to about 60 μM, about 20 nM to about 55 μM, about 20 nM to about 50 μM, about 20 nM to about 45 μM, about 20 nM to about 40 μM, about 20 nM to about 45 μM, about 20 nM to about 40 μM, about 20 nM to about 35 μM, about 20 nM to about 30 μM, about 20 nM to about 25 μM, about 20 nM to about 20 μM, about 20 nM to about 15 , about 20nM to about 10 μM, about 20nM to about 5 μM, about 20nM to about 2 μM, about 20nM to about 1 μM, about 20nM to about 950nM, about 20nM to about 900nM, about 20nM to about 850nM, about 20nM to about 800nM, about 20nM to about 750nM, about 20nM to about 700nM, about 20nM to about 650nM, about 20nM to about 600nM, about 20nM to about 550nM, about 20nM to about 500nM, about 20nM to about 450nM, about 20nM to about 400nM, about 20nM to about 350nM, about 20nM to about 300nM, about 20nM to about 250nM, about 20nM to about about 20nM to about 150nM, about 20nM to about 100nM, about 20nM to about 95nM, about 20nM to about 90nM, about 20nM to about 85nM, about 20nM to about 80nM, about 20nM to about 75nM, about 20nM to about 70nM, about 20nM to about 75nM, about 20nM to about 70nM, about 20nM to about 70nM, about 20nM to about 65nM, about 20nM to about 60nM, about 20nM to about 55nM, about 20nM to about 50nM, about 20nM to about 45nM, about 20nM to about 40nM, about 20nM to about 35nM, about 20nM to about 30nM, about 20nM to about 25nM, about 50nM to about 100μM, about 50nM to about 95μM, about 50nM to about 90μM, about 50nM to about 85μM, about 50nM to about 80μM, about 50nM to about 75μM, about 50nM to about 70μM, about 50nM to about 65μM, about 50nM to about 60μM, about 50nM to about 55μM, about 50nM to about 50μM, about 50nM to about 45μM, about 50nM to about 40μM, about 50nM to about 45μM, about 50nM to about 40μM, about 50nM to about 40μM, about 50nM to about 35μM, about 50nM to about 30μM, about 50nM to about 25μM, about 50nM to about 20μM, about 50nM to about 15μM, about 50nM to about 10μM, about 50nM to about 5μM,about 50nM to about 2μM, about 50nM to about 1μM, about 50nM to about 950nM, about 50nM to about 900nM, about 50nM to about 850nM, about 50nM to about 800nM, about 50nM to about 750nM, about 50nM to about 700nM, about 50nM to about 650nM, about 50nM to about 600nM, about 50nM to about 550nM, about 50nM to about 500nM, about 50nM to about 450nM, about 50nM to about 400nM, about 50nM to about 350nM, about 50nM to about 300nM, about 50nM to about 250nM, about 50nM to about 200nM, about 50nM to about 150nM, about 50nM to about M to about 100 nM, about 50 nM to about 95 nM, about 50 nM to about 90 nM, about 50 nM to about 85 nM, about 50 nM to about 80 nM, about 50 nM to about 75 nM, about 50 nM to about 70 nM, about 50 nM to about 75 nM, about 50 nM to about 70 nM, about 50 nM to about 65 nM, about 50 nM to about 60 nM, about 50 nM to about 55 nM, about 100 nM to about 100 μM, about 100 nM to about 95 μM, about 100 nM to about 90 μM, about 100 nM to about 85 μM, about 100 nM to about 80 μM, about 100 nM to about 75 μM, about 100 nM to about 70 μM, about 100 nM to about 65 μM, about 100nM to about 60μM, about 100nM to about 55μM, about 100nM to about 50μM, about 100nM to about 45μM, about 100nM to about 40μM, about 100nM to about 45μM, about 100nM to about 40μM, about 100nM to about 35μM, about 100nM to about 30μM, about 100nM to about 25μM, about 100nM to about 20μM, about 100nM to about 15μM, about 100nM to about 10μM, about 100nM to about 5μM, about 100nM to about 2μM, about 100nM to about 1μM, about 100nM to about 950nM, about 100nM to about 900nM, about 100nM to about 850nM, about 100nM to about 800nM, about 100nM to about 750nM, about 100nM to about 700nM, about 100nM to about 650nM, about 100nM to about 600nM, about 100nM to about 550nM, about 100nM to about 500nM, about 100nM to about 450nM, about 100nM to about 400nM, about 100nM to about 350nM, about 100nM to about 300nM, about 100nM to about 250nM, about 100nM to about 200nM, about 100nM to about 150nM, about 200nM to about 100μM, about 200nM to about 95μM, about 200nM to about 90μM, about 200nM to about 85μM,about 200nM to about 80μM, about 200nM to about 75μM, about 200nM to about 70μM, about 200nM to about 65μM, about 200nM to about 60μM, about 200nM to about 55μM, about 200nM to about 50μM, about 200nM to about 45μM, about 200nM to about 40μM, about 200nM to about 45μM, about 200nM to about 40μM, about 200nM to about 40μM, about 200nM to about 35μM, about 200nM to about 30μM, about 200nM to about 25μM, about 200nM to about 20μM, about 200nM to about 15μM, about 200nM to about 10μM, about 200nM to about 5μM, about 200nM to about 2μM, about 200 nM to about 1 μM, about 200 nM to about 950 nM, about 200 nM to about 900 nM, about 200 nM to about 850 nM, about 200 nM to about 800 nM, about 200 nM to about 750 nM, about 200 nM to about 700 nM, about 200 nM to about 650 nM, about 200 nM to about 600 nM, about 200 nM to about 550 nM, about 200 nM to about 500 nM, about 200 nM to about 450 nM, about 200 nM to about 400 nM, about 200 nM to about 350 nM, about 200 nM to about 300 nM, about 200 nM to about 250 nM, about 300 nM to about 100 μM, about 300 nM to about 95 μM, about 300 nM to about In some embodiments, the present invention relates to a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, a method for preparing an aqueous solution of at least 200 μM aqueous solution, about 300nM to about 1 μM, about 300nM to about 950nM, about 300nM to about 900nM, about 300nM to about 850nM, about 300nM to about 800nM, about 300nM to about 750nM, about 300nM to about 700nM, about 300nM to about 650nM, about 300nM to about 600nM, about 300nM to about 550nM, about 300nM to about 500nM, about 300nM to about 450nM, about 300nM to about 400nM, about 300nM to about 350nM, about 400nM to about 100μM, about 400nM to about 95μM, about 400nM to about 90μM,about 400nM to about 85μM, about 400nM to about 80μM, about 400nM to about 75μM, about 400nM to about 70μM, about 400nM to about 65μM, about 400nM to about 60μM, about 400nM to about 55μM, about 400nM to about 50μM, about 400nM to about 45μM, about 400nM to about 40μM, about 400nM to about 45μM, about 400nM to about 40μM, about 400nM to about 40μM, about 400nM to about 35μM, about 400nM to about 30μM, about 400nM to about 25μM, about 400nM to about 20μM, about 400nM to about 15μM, about 400nM to about 10μM, about 400nM to about 5μM, about 400nM to about 2μM, about 400nM to about 1μM, about 400nM to about 950nM, about 400nM to about 900nM, about 400nM to about 850nM, about 400nM to about 800nM, about 400nM to about 750nM, about 400nM to about 700nM, about 400nM to about 650nM, about 400nM to about 600nM, about 400nM to about 550nM, about 400nM to about 500nM, about 400nM to about 450nM, about 500nM to about 100μM, about 500nM to about 95μM, about 500nM to about 90μM, about 500nM to about 85μM, about 500nM to about 80μM, about 500nM to about In some embodiments, the present invention relates to a polymerizable substance of the present invention and / or a polymerizable substance of the present invention. The polymerizable substance of the present invention is preferably a polymerizable substance of the present invention, wherein the polymerizable substance is a polymerizable substance of the present invention and / or a polymerizable substance of the present invention. In some embodiments, the polymerizable substance is preferably a polymerizable substance of the present invention and / or a polymerizable substance of the present invention. In some embodiments, the present invention relates to a pharmaceutical composition comprising the following: about 500nM to about 900nM, about 500nM to about 850nM, about 500nM to about 800nM, about 500nM to about 750nM, about 500nM to about 700nM, about 500nM to about 650nM, about 500nM to about 600nM, about 500nM to about 550nM, about 600nM to about 100μM, about 600nM to about 95μM, about 600nM to about 90μM, about 600nM to about 85μM, about 600nM to about 80μM, about 600nM to about 75μM, about 600nM to about 70μM, about 600nM to about 65μM, about 600nM to about 60μM, about 600nM to about 55μM,about 600nM to about 50μM, about 600nM to about 45μM, about 600nM to about 40μM, about 600nM to about 45μM, about 600nM to about 40μM, about 600nM to about 35μM, about 600nM to about 30μM, about 600nM to about 25μM, about 600nM to about 20μM, about 600nM to about 15μM, about 600nM to about 10μM, about 600nM to about 5μM, about 600nM to about 2μM, about 600nM to about 1μM, about 600nM to about 950nM, about 600nM to about 900nM, about 600nM to about 850nM, about 600nM to about 800nM, about 600nM to about 750nM , about 600nM to about 700nM, about 600nM to about 650nM, about 700nM to about 100μM, about 700nM to about 95μM, about 700nM to about 90μM, about 700nM to about 85μM, about 700nM to about 80μM, about 700nM to about 75μM, about 700nM to about 70μM, about 700nM to about 65μM, about 700nM to about 60μM, about 700nM to about 55μM, about 700nM to about 50μM, about 700nM to about 45μM, about 700nM to about 40μM, about 700nM to about 45μM, about 700nM to about 40μM, about 700nM to about 35μM, about 700nM to about 30μM In some embodiments, the present invention relates to about 700 nM to about 25 μM, about 700 nM to about 20 μM, about 700 nM to about 15 μM, about 700 nM to about 10 μM, about 700 nM to about 5 μM, about 700 nM to about 2 μM, about 700 nM to about 1 μM, about 700 nM to about 950 nM, about 700 nM to about 900 nM, about 700 nM to about 850 nM, about 700 nM to about 800 nM, about 700 nM to about 750 nM, about 800 nM to about 100 μM, about 800 nM to about 95 μM, about 800 nM to about 90 μM, about 800 nM to about 85 μM, about 800 nM to about 80 μM, about 800 nM to about 75 μM, about 800 nM to about 70 In some embodiments, the present invention relates to a novel nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam, a nanostructured carbon foam,about 800nM to about 850nM, about 900nM to about 100μM, about 900nM to about 95μM, about 900nM to about 90μM, about 900nM to about 85μM, about 900nM to about 80μM, about 900nM to about 75μM, about 900nM to about 70μM, about 900nM to about 65μM, about 900nM to about 60μM, about 900nM to about 55μM, about 900nM to about 50μM, about 900nM to about 45μM, about 900nM to about 40μM, about 900nM to about 45μM, about 900nM to about 40μM, about 900nM to about 35μM, about 900nM to about 30μM, about 900nM to about 25μM, about 900nM to about 20μM, about 900nM to about 15μM, about 900nM to about 10μM, about 900nM to about 5μM, about 900nM to about 2μM, about 900nM to about 1μM, about 900nM to about 950nM, about 1μM to about 100μM, about 1μM to about 95μM, about 1μM to about 90μM, about 1μM to about 85μM, about 1μM to about 80μM, about 1μM to about 75μM, about 1μM to about 70μM, about 1μM to about 65μM, about 1μM to about 60μM, about 1μM to about 55μM, about 1μM to about 50μM, about 1μM to about 45μM, about 1μM to about 40μM, about 1μM to about 45μM, about 1μM to about 40μM, about 1μM to about 35μM, about 1μM to about 30μM, about 1μM to about 25μM, about 1μM to about 20μM, about 1μM to about 15μM, about 1μM to about 10μM, about 1μM to about 5μM, about 1μM to about 2μM, about 2μM to about 100μM, about 2μM to about 95μM, about 2μM to about 90μM, about 2μM to about 85μM, about 2μM to about 80μM, about 2μM to about 75μM, about 2μM to about 70μM, about 2μM to about 65μM, about 2μM to about 60μM, about 2μM to about 55μM, about 2μM to about 50μM, about 2μM to about 45μM, about 2μM to about 40μM, about 2μM to about 45μM, about 2μM to about M to about 40 μM, about 2 μM to about 35 μM, about 2 μM to about 30 μM, about 2 μM to about 25 μM, about 2 μM to about 20 μM, about 2 μM to about 15 μM, about 2 μM to about 10 μM, about 2 μM to about 5 μM, about 5 μM to about 100 μM, about 5 μM to about 95 μM, about 5 μM to about 90 μM, about 5 μM to about 85 μM, about 5 μM to about 80 μM, about 5 μM to about 75 μM, about 5 μM to about 70 μM, about 5 μM to about 65 μM, about 5 μM to about 60 μM, about 5 μM to about 55 μM, about 5 μM to about 50 μM, about 5 μM to about 45 μM, about 5 μM to about 40 μM, about 5 μM to about 45 μM, about 5 μM to about 40 μM,about 5 μM to about 35 μM, about 5 μM to about 30 μM, about 5 μM to about 25 μM, about 5 μM to about 20 μM, about 5 μM to about 15 μM, about 5 μM to about 10 μM, about 10 μM to about 100 μM, about 10 μM to about 95 μM, about 10 μM to about 90 μM, about 10 μM to about 85 μM, about 10 μM to about 80 μM, about 10 μM to about 75 μM, about 10 μM to about 70 μM, about 10 μM to about 65 μM, about 10 μM to about 60 μM, about 10 μM to about 55 μM, about 10 μM to about 50 μM, about 10 μM to about 45 μM, about 10 μM to about 40 μM, about 10 μM to about 45 μM, about 10 μM to about 40 μM, about 1 0μM to about 35μM, about 10μM to about 30μM, about 10μM to about 25μM, about 10μM to about 20μM, about 10μM to about 15μM, about 20μM to about 100μM, about 20μM to about 95μM, about 20μM to about 90μM, about 20μM to about 85μM, about 20μM to about 80μM, about 20μM to about 75μM, about 20μM to about 70μM, about 20μM to about 65μM, about 20μM to about 60μM, about 20μM to about 55μM, about 20μM to about 50μM, about 20μM to about 45μM, about 20μM to about 40μM, about 20μM to about 45μM, about 20μM to about 40μM, about 20μM to about 35μM M, about 20 μM to about 30 μM, about 20 μM to about 25 μM, about 30 μM to about 100 μM, about 30 μM to about 95 μM, about 30 μM to about 90 μM, about 30 μM to about 85 μM, about 30 μM to about 80 μM, about 30 μM to about 75 μM, about 30 μM to about 70 μM, about 30 μM to about 65 μM, about 30 μM to about 60 μM, about 30 μM to about 55 μM, about 30 μM to about 50 μM, about 30 μM to about 45 μM, about 30 μM to about 40 μM, about 30 μM to about 45 μM, about 30 μM to about 40 μM, about 30 μM to about 35 μM, about 40 μM to about 100 μM, about 40 μM to about 95 μM, about 40 μM about 40 μM to about 85 μM, about 40 μM to about 80 μM, about 40 μM to about 75 μM, about 40 μM to about 70 μM, about 40 μM to about 65 μM, about 40 μM to about 60 μM, about 40 μM to about 55 μM, about 40 μM to about 50 μM, about 40 μM to about 45 μM, about 50 μM to about 100 μM, about 50 μM to about 95 μM, about 50 μM to about 90 μM, about 50 μM to about 85 μM, about 50 μM to about 80 μM, about 50 μM to about 75 μM, about 50 μM to about 70 μM, about 50 μM to about 65 μM, about 50 μM to about 60 μM, about 50 μM to about 55 μM, about 60 μM to about 100 μM,The dissociation equilibrium constant (K, K) of about 60 μM to about 95 μM, about 60 μM to about 90 μM, about 60 μM to about 85 μM, about 60 μM to about 80 μM, about 60 μM to about 75 μM, about 60 μM to about 70 μM, about 60 μM to about 65 μM, about 70 μM to about 100 μM, about 70 μM to about 95 μM, about 70 μM to about 90 μM, about 70 μM to about 85 μM, about 70 μM to about 80 μM, about 70 μM to about 75 μM, about 80 μM to about 100 μM, about 80 μM to about 95 μM, about 80 μM to about 90 μM, about 80 μM to about 85 μM, about 90 μM to about 100 μM, or about 90 μM to about 95 μM. d ) is bound to β-TCP (e.g., any type of β-TCP described herein) (e.g., as measured using SPR in phosphate-buffered saline).

[0190] In some embodiments, the chimeric polypeptides described herein can have from about 20 amino acids to about 1400 amino acids, from about 20 amino acids to about 1350 amino acids, from about 20 amino acids to about 1300 amino acids, from about 20 amino acids to about 1250 amino acids, from about 20 amino acids to about 1200 amino acids, from about 20 amino acids to about 1150 amino acids, from about 20 amino acids to about 1100 amino acids, from about 20 amino acids to about 1050 amino acids, from about 20 amino acids to about 1000 amino acids, from about 20 amino acids to about 950 amino acids, from about 20 amino acids to about 900 amino acids, from about 20 amino acids to about 850 amino acids, from about 20 amino acids to about 800 amino acids. acid, about 20 amino acids to about 750 amino acids, about 20 amino acids to about 700 amino acids, about 20 amino acids to about 650 amino acids, about 20 amino acids to about 600 amino acids, about 20 amino acids to about 550 amino acids, about 20 amino acids to about 500 amino acids, about 20 amino acids to about 450 amino acids, about 20 amino acids to about 400 amino acids, about 20 amino acids to about 350 amino acids, about 20 amino acids to about 300 amino acids, about 20 amino acids to about 250 amino acids, about 20 amino acids to about 200 amino acids, about 20 amino acids to about 150 amino acids, about 20 amino acids to about 100 amino acids, about 20 amino acids to about 9 0 amino acids, about 20 amino acids to about 80 amino acids, about 20 amino acids to about 70 amino acids, about 20 amino acids to about 60 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 45 amino acids, about 20 amino acids to about 40 amino acids, about 20 amino acids to about 35 amino acids, about 20 amino acids to about 30 amino acids, about 20 amino acids to about 25 amino acids, about 25 amino acids to about 1400 amino acids, about 25 amino acids to about 1350 amino acids, about 25 amino acids to about 1300 amino acids, about 25 amino acids to about 1250 amino acids, about 25 amino acids to about 1200 amino acids, about 25 amino acids to about about 25 amino acids to about 1150 amino acids, about 25 amino acids to about 1100 amino acids, about 25 amino acids to about 1050 amino acids, about 25 amino acids to about 1000 amino acids, about 25 amino acids to about 950 amino acids, about 25 amino acids to about 900 amino acids, about 25 amino acids to about 850 amino acids, about 25 amino acids to about 800 amino acids, about 25 amino acids to about 750 amino acids, about 25 amino acids to about 700 amino acids, about 25 amino acids to about 650 amino acids, about 25 amino acids to about 600 amino acids, about 25 amino acids to about 550 amino acids, about 25 amino acids to about 500 amino acids, about 25 amino acids to about 450 amino acids,about 25 amino acids to about 400 amino acids, about 25 amino acids to about 350 amino acids, about 25 amino acids to about 300 amino acids, about 25 amino acids to about 250 amino acids, about 25 amino acids to about 200 amino acids, about 25 amino acids to about 150 amino acids, about 25 amino acids to about 100 amino acids, about 25 amino acids to about 90 amino acids, about 25 amino acids to about 80 amino acids, about 25 amino acids to about 70 amino acids, about 25 amino acids to about 60 amino acids, about 25 amino acids to about 50 amino acids, about 25 amino acids to about 45 amino acids, about 25 amino acids to about 40 amino acids, about 25 amino acids to about 35 amino acids. , about 25 amino acids to about 30 amino acids, about 30 amino acids to about 1400 amino acids, about 30 amino acids to about 1350 amino acids, about 30 amino acids to about 1300 amino acids, about 30 amino acids to about 1250 amino acids, about 30 amino acids to about 1200 amino acids, about 30 amino acids to about 1150 amino acids, about 30 amino acids to about 1100 amino acids, about 30 amino acids to about 1050 amino acids, about 30 amino acids to about 1000 amino acids, about 30 amino acids to about 950 amino acids, about 30 amino acids to about 900 amino acids, about 30 amino acids to about 850 amino acids, about 30 amino acids to about 800 amino acids , about 30 amino acids to about 750 amino acids, about 30 amino acids to about 700 amino acids, about 30 amino acids to about 650 amino acids, about 30 amino acids to about 600 amino acids, about 30 amino acids to about 550 amino acids, about 30 amino acids to about 500 amino acids, about 30 amino acids to about 450 amino acids, about 30 amino acids to about 400 amino acids, about 30 amino acids to about 350 amino acids, about 30 amino acids to about 300 amino acids, about 30 amino acids to about 250 amino acids, about 30 amino acids to about 200 amino acids, about 30 amino acids to about 150 amino acids, about 30 amino acids to about 100 amino acids, about 30 amino acids about 30 amino acids to about 80 amino acids, about 30 amino acids to about 70 amino acids, about 30 amino acids to about 60 amino acids, about 30 amino acids to about 50 amino acids, about 30 amino acids to about 45 amino acids, about 30 amino acids to about 40 amino acids, about 30 amino acids to about 35 amino acids, about 35 amino acids to about 1400 amino acids, about 35 amino acids to about 1350 amino acids, about 35 amino acids to about 1300 amino acids, about 35 amino acids to about 1250 amino acids, about 35 amino acids to about 1200 amino acids, about 35 amino acids to about 1150 amino acids, about 35 amino acids to about 1100 amino acids,about 35 amino acids to about 1050 amino acids, about 35 amino acids to about 1000 amino acids, about 35 amino acids to about 950 amino acids, about 35 amino acids to about 900 amino acids, about 35 amino acids to about 850 amino acids, about 35 amino acids to about 800 amino acids, about 35 amino acids to about 750 amino acids, about 35 amino acids to about 700 amino acids, about 35 amino acids to about 650 amino acids, about 35 amino acids to about 600 amino acids, about 35 amino acids to about 550 amino acids, about 35 amino acids to about 500 amino acids, about 35 amino acids to about 450 amino acids, about 35 amino acids to about 400 amino acids, about 35 amino acids to about 5 about 35 amino acids to about 350 amino acids, about 35 amino acids to about 300 amino acids, about 35 amino acids to about 250 amino acids, about 35 amino acids to about 200 amino acids, about 35 amino acids to about 150 amino acids, about 35 amino acids to about 100 amino acids, about 35 amino acids to about 90 amino acids, about 35 amino acids to about 80 amino acids, about 35 amino acids to about 70 amino acids, about 35 amino acids to about 60 amino acids, about 35 amino acids to about 50 amino acids, about 35 amino acids to about 45 amino acids, about 35 amino acids to about 40 amino acids, about 40 amino acids to about 1400 amino acids, about 40 amino acids to about 1350 amino acids, about 40 about 40 amino acids to about 1300 amino acids, about 40 amino acids to about 1250 amino acids, about 40 amino acids to about 1200 amino acids, about 40 amino acids to about 1150 amino acids, about 40 amino acids to about 1100 amino acids, about 40 amino acids to about 1050 amino acids, about 40 amino acids to about 1000 amino acids, about 40 amino acids to about 950 amino acids, about 40 amino acids to about 900 amino acids, about 40 amino acids to about 850 amino acids, about 40 amino acids to about 800 amino acids, about 40 amino acids to about 750 amino acids, about 40 amino acids to about 700 amino acids, about 40 amino acids to about 650 amino acids, about 40 from about 40 amino acids to about 600 amino acids, from about 40 amino acids to about 550 amino acids, from about 40 amino acids to about 500 amino acids, from about 40 amino acids to about 450 amino acids, from about 40 amino acids to about 400 amino acids, from about 40 amino acids to about 350 amino acids, from about 40 amino acids to about 300 amino acids, from about 40 amino acids to about 250 amino acids, from about 40 amino acids to about 200 amino acids, from about 40 amino acids to about 150 amino acids, from about 40 amino acids to about 100 amino acids, from about 40 amino acids to about 90 amino acids, from about 40 amino acids to about 80 amino acids, from about 40 amino acids to about 70 amino acids, from about 40 amino acids to about 60 amino acids,about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 45 amino acids to about 1400 amino acids, about 45 amino acids to about 1350 amino acids, about 45 amino acids to about 1300 amino acids, about 45 amino acids to about 1250 amino acids, about 45 amino acids to about 1200 amino acids, about 45 amino acids to about 1150 amino acids, about 45 amino acids to about 1100 amino acids, about 45 amino acids to about 1050 amino acids, about 45 amino acids to about 1000 amino acids, about 45 amino acids to about 950 amino acids, about 45 amino acids to about 900 amino acids, about 45 amino acids to about 850 amino acids, about 45 In some embodiments, the present invention relates to a polypeptide having at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. about 45 amino acids to about 90 amino acids, about 45 amino acids to about 80 amino acids, about 45 amino acids to about 70 amino acids, about 45 amino acids to about 60 amino acids, about 45 amino acids to about 50 amino acids, about 50 amino acids to about 1400 amino acids, about 50 amino acids to about 1350 amino acids, about 50 amino acids to about 1300 amino acids, about 50 amino acids to about 1250 amino acids, about 50 amino acids to about 1200 amino acids, about 50 amino acids to about 1150 amino acids, about 50 amino acids to about 1100 amino acids, about 50 amino acids to about 1050 amino acids, about 50 amino acids to about 1000 amino acids, about 50 amino acids to about 1500 amino acids, about 50 amino acids to about 1600 amino acids, about 50 amino acids to about 1700 amino acids, about 50 amino acids to about 1800 amino acids, about 50 amino acids to about 1900 amino acids, about 50 amino acids to about 1900 amino acids, about 50 amino acids to about 1800 amino acids, about 50 amino acids to about 1900 amino acids, about 50 amino acids to about 1900 amino acids about 50 amino acids to about 950 amino acids, about 50 amino acids to about 900 amino acids, about 50 amino acids to about 850 amino acids, about 50 amino acids to about 800 amino acids, about 50 amino acids to about 750 amino acids, about 50 amino acids to about 700 amino acids, about 50 amino acids to about 650 amino acids, about 50 amino acids to about 600 amino acids, about 50 amino acids to about 550 amino acids, about 50 amino acids to about 500 amino acids, about 50 amino acids to about 450 amino acids, about 50 amino acids to about 400 amino acids, about 50 amino acids to about 350 amino acids, about 50 amino acids to about 300 amino acids, about 50 amino acids to about 250 amino acids,from about 50 amino acids to about 200 amino acids, from about 50 amino acids to about 150 amino acids, from about 50 amino acids to about 100 amino acids, from about 50 amino acids to about 90 amino acids, from about 50 amino acids to about 80 amino acids, from about 50 amino acids to about 70 amino acids, from about 50 amino acids to about 60 amino acids, from about 60 amino acids to about 1400 amino acids, from about 60 amino acids to about 1350 amino acids, from about 60 amino acids to about 1300 amino acids, from about 60 amino acids to about 1250 amino acids, from about 60 amino acids to about 1200 amino acids, from about 60 amino acids to about 1150 amino acids, from about 60 amino acids to about 1100 amino acids, from about 60 amino acids to about about 1050 amino acids, about 60 amino acids to about 1000 amino acids, about 60 amino acids to about 950 amino acids, about 60 amino acids to about 900 amino acids, about 60 amino acids to about 850 amino acids, about 60 amino acids to about 800 amino acids, about 60 amino acids to about 750 amino acids, about 60 amino acids to about 700 amino acids, about 60 amino acids to about 650 amino acids, about 60 amino acids to about 600 amino acids, about 60 amino acids to about 550 amino acids, about 60 amino acids to about 500 amino acids, about 60 amino acids to about 450 amino acids, about 60 amino acids to about 400 amino acids, about 60 amino acids to about 350 amino acids, from about 60 amino acids to about 300 amino acids, from about 60 amino acids to about 250 amino acids, from about 60 amino acids to about 200 amino acids, from about 60 amino acids to about 150 amino acids, from about 60 amino acids to about 100 amino acids, from about 60 amino acids to about 90 amino acids, from about 60 amino acids to about 80 amino acids, from about 60 amino acids to about 70 amino acids, from about 70 amino acids to about 1400 amino acids, from about 70 amino acids to about 1350 amino acids, from about 70 amino acids to about 1300 amino acids, from about 70 amino acids to about 1250 amino acids, from about 70 amino acids to about 1200 amino acids, from about 70 amino acids to about 1150 amino acids, from about 70 amino acids to about 1500 amino acids, about 70 amino acids to about 1050 amino acids, about 70 amino acids to about 1000 amino acids, about 70 amino acids to about 950 amino acids, about 70 amino acids to about 900 amino acids, about 70 amino acids to about 850 amino acids, about 70 amino acids to about 800 amino acids, about 70 amino acids to about 750 amino acids, about 70 amino acids to about 700 amino acids, about 70 amino acids to about 650 amino acids, about 70 amino acids to about 600 amino acids, about 70 amino acids to about 550 amino acids, about 70 amino acids to about 500 amino acids, about 70 amino acids to about 450 amino acids, about 70 amino acids to about 400 amino acids,from about 70 amino acids to about 350 amino acids, from about 70 amino acids to about 300 amino acids, from about 70 amino acids to about 250 amino acids, from about 70 amino acids to about 200 amino acids, from about 70 amino acids to about 150 amino acids, from about 70 amino acids to about 100 amino acids, from about 70 amino acids to about 90 amino acids, from about 70 amino acids to about 80 amino acids, from about 80 amino acids to about 1400 amino acids, from about 80 amino acids to about 1350 amino acids, from about 80 amino acids to about 1300 amino acids, from about 80 amino acids to about 1250 amino acids, from about 80 amino acids to about 1200 amino acids, from about 80 amino acids to about 1150 amino acids, from about 80 amino acids to about 1500 amino acids, about 1100 amino acids, about 80 amino acids to about 1050 amino acids, about 80 amino acids to about 1000 amino acids, about 80 amino acids to about 950 amino acids, about 80 amino acids to about 900 amino acids, about 80 amino acids to about 850 amino acids, about 80 amino acids to about 800 amino acids, about 80 amino acids to about 750 amino acids, about 80 amino acids to about 700 amino acids, about 80 amino acids to about 650 amino acids, about 80 amino acids to about 600 amino acids, about 80 amino acids to about 550 amino acids, about 80 amino acids to about 500 amino acids, about 80 amino acids to about 450 amino acids, about 80 amino acids to about 400 amino acids, about 80 amino acids to about 350 amino acids, about 80 amino acids to about 300 amino acids, about 80 amino acids to about 250 amino acids, about 80 amino acids to about 200 amino acids, about 80 amino acids to about 150 amino acids, about 80 amino acids to about 100 amino acids, about 80 amino acids to about 90 amino acids, about 90 amino acids to about 1400 amino acids, about 90 amino acids to about 1350 amino acids, about 90 amino acids to about 1300 amino acids, about 90 amino acids to about 1250 amino acids, about 90 amino acids to about 1200 amino acids, about 90 amino acids to about 1150 amino acids, about 90 amino acids to about 1100 amino acids, about 90 amino acids to about In some embodiments, the present invention relates to a polypeptide having a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight of 500 amino acids, a molecular weight ofFrom about 90 amino acids to about 300 amino acids, from about 90 amino acids to about 250 amino acids, from about 90 amino acids to about 200 amino acids, from about 90 amino acids to about 150 amino acids, from about 90 amino acids to about 100 amino acids, from about 100 amino acids to about 1400 amino acids, from about 100 amino acids to about 1350 amino acids, from about 100 amino acids to about 1300 amino acids, from about 100 amino acids to about 1250 amino acids, from about 100 amino acids to about 1200 amino acids, from about 100 amino acids to about 1150 amino acids, from about 100 amino acids to about 1100 amino acids, from about 100 amino acids to about 1050 amino acids, from about 100 amino acids to about 1 about 1000 amino acids, about 100 amino acids to about 950 amino acids, about 100 amino acids to about 900 amino acids, about 100 amino acids to about 850 amino acids, about 100 amino acids to about 800 amino acids, about 100 amino acids to about 750 amino acids, about 100 amino acids to about 700 amino acids, about 100 amino acids to about 650 amino acids, about 100 amino acids to about 600 amino acids, about 100 amino acids to about 550 amino acids, about 100 amino acids to about 500 amino acids, about 100 amino acids to about 450 amino acids, about 100 amino acids to about 400 amino acids, about 100 amino acids to about 350 amino acids, about 1 00 amino acids to about 300 amino acids, about 100 amino acids to about 250 amino acids, about 100 amino acids to about 200 amino acids, about 100 amino acids to about 150 amino acids, about 150 amino acids to about 1400 amino acids, about 150 amino acids to about 1350 amino acids, about 150 amino acids to about 1300 amino acids, about 150 amino acids to about 1250 amino acids, about 150 amino acids to about 1200 amino acids, about 150 amino acids to about 1150 amino acids, about 150 amino acids to about 1100 amino acids, about 150 amino acids to about 1050 amino acids, about 150 amino acids to about 1000 amino acids, about 150 from about 150 amino acids to about 950 amino acids, from about 150 amino acids to about 900 amino acids, from about 150 amino acids to about 850 amino acids, from about 150 amino acids to about 800 amino acids, from about 150 amino acids to about 750 amino acids, from about 150 amino acids to about 700 amino acids, from about 150 amino acids to about 650 amino acids, from about 150 amino acids to about 600 amino acids, from about 150 amino acids to about 550 amino acids, from about 150 amino acids to about 500 amino acids, from about 150 amino acids to about 450 amino acids, from about 150 amino acids to about 400 amino acids, from about 150 amino acids to about 350 amino acids, from about 150 amino acids to about 300 amino acids,about 150 amino acids to about 250 amino acids, about 150 amino acids to about 200 amino acids, about 200 amino acids to about 1400 amino acids, about 200 amino acids to about 1350 amino acids, about 200 amino acids to about 1300 amino acids, about 200 amino acids to about 1250 amino acids, about 200 amino acids to about 1200 amino acids, about 200 amino acids to about 1150 amino acids, about 200 amino acids to about 1100 amino acids, about 200 amino acids to about 1050 amino acids, about 200 amino acids to about 1000 amino acids, about 200 amino acids to about 950 amino acids, about 200 amino acids to about 900 amino acids, about 200 amino acids to about 850 amino acids, about 200 amino acids to about 800 amino acids, about 200 amino acids to about 750 amino acids, about 200 amino acids to about 700 amino acids, about 200 amino acids to about 650 amino acids, about 200 amino acids to about 600 amino acids, about 200 amino acids to about 550 amino acids, about 200 amino acids to about 500 amino acids, about 200 amino acids to about 450 amino acids, about 200 amino acids to about 400 amino acids, about 200 amino acids to about 350 amino acids, about 200 amino acids to about 300 amino acids, about 200 amino acids to about 250 amino acids, about 250 amino acids to about 1400 amino acids, about from about 250 amino acids to about 1350 amino acids, from about 250 amino acids to about 1300 amino acids, from about 250 amino acids to about 1250 amino acids, from about 250 amino acids to about 1200 amino acids, from about 250 amino acids to about 1150 amino acids, from about 250 amino acids to about 1100 amino acids, from about 250 amino acids to about 1050 amino acids, from about 250 amino acids to about 1000 amino acids, from about 250 amino acids to about 950 amino acids, from about 250 amino acids to about 900 amino acids, from about 250 amino acids to about 850 amino acids, from about 250 amino acids to about 800 amino acids, from about 250 amino acids to about 750 amino acids, from about 250 amino acids to about 1 about 250 amino acids to about 700 amino acids, about 250 amino acids to about 650 amino acids, about 250 amino acids to about 600 amino acids, about 250 amino acids to about 550 amino acids, about 250 amino acids to about 500 amino acids, about 250 amino acids to about 450 amino acids, about 250 amino acids to about 400 amino acids, about 250 amino acids to about 350 amino acids, about 250 amino acids to about 300 amino acids, about 300 amino acids to about 1400 amino acids, about 300 amino acids to about 1350 amino acids, about 300 amino acids to about 1300 amino acids, about 300 amino acids to about 1250 amino acids, about 300 amino acids to about 1200 amino acids,about 300 amino acids to about 1150 amino acids, about 300 amino acids to about 1100 amino acids, about 300 amino acids to about 1050 amino acids, about 300 amino acids to about 1000 amino acids, about 300 amino acids to about 950 amino acids, about 300 amino acids to about 900 amino acids, about 300 amino acids to about 850 amino acids, about 300 amino acids to about 800 amino acids, about 300 amino acids to about 750 amino acids, about 300 amino acids to about 700 amino acids, about 300 amino acids to about 650 amino acids, about 300 amino acids to about 600 amino acids, about 300 amino acids to about 550 amino acids, about 300 amino acids to about The present invention relates to a polypeptide having a molecular weight of 500 amino acids, about 300 amino acids to about 450 amino acids, about 300 amino acids to about 400 amino acids, about 300 amino acids to about 350 amino acids, about 350 amino acids to about 1400 amino acids, about 350 amino acids to about 1350 amino acids, about 350 amino acids to about 1300 amino acids, about 350 amino acids to about 1250 amino acids, about 350 amino acids to about 1200 amino acids, about 350 amino acids to about 1150 amino acids, about 350 amino acids to about 1100 amino acids, about 350 amino acids to about 1050 amino acids, about 350 amino acids to about 1000 amino acids, about 350 amino acids to about 950 amino acids. acid, about 350 amino acids to about 900 amino acids, about 350 amino acids to about 850 amino acids, about 350 amino acids to about 800 amino acids, about 350 amino acids to about 750 amino acids, about 350 amino acids to about 700 amino acids, about 350 amino acids to about 650 amino acids, about 350 amino acids to about 600 amino acids, about 350 amino acids to about 550 amino acids, about 350 amino acids to about 500 amino acids, about 350 amino acids to about 450 amino acids, about 350 amino acids to about 400 amino acids, about 400 amino acids to about 1400 amino acids, about 400 amino acids to about 1350 amino acids, about 400 amino acids to about about 400 amino acids to about 900 amino acids, about 400 amino acids to about 850 amino acids, about 400 amino acids to about 800 amino acids, about 400 amino acids to about 750 amino acids, about 400 amino acids to about 700 amino acids, about 400 amino acids to about 650 amino acids, about 400 amino acids to about 1000 amino acids, about 400 amino acids to about 1050 amino acids, about 400 amino acids to about 1000 amino acids, about 400 amino acids to about 950 amino acids, about 400 amino acids to about 900 amino acids, about 400 amino acids to about 850 amino acids, about 400 amino acids to about 800 amino acids, about 400 amino acids to about 750 amino acids, about 400 amino acids to about 700 amino acids, about 400 amino acids to about 650 amino acids,about 400 amino acids to about 600 amino acids, about 400 amino acids to about 550 amino acids, about 400 amino acids to about 500 amino acids, about 400 amino acids to about 450 amino acids, about 450 amino acids to about 1400 amino acids, about 450 amino acids to about 1350 amino acids, about 450 amino acids to about 1300 amino acids, about 450 amino acids to about 1250 amino acids, about 450 amino acids to about 1200 amino acids, about 450 amino acids to about 1150 amino acids, about 450 amino acids to about 1100 amino acids, about 450 amino acids to about 1050 amino acids, about 450 amino acids to about 1000 amino acids, about 450 amino acids to about 1500 amino acids, about 450 amino acids to about 1600 amino acids, about 450 amino acids to about 1700 amino acids, about 450 amino acids to about 1800 amino acids, about 450 amino acids to about 19 ... about 450 amino acids to about 950 amino acids, about 450 amino acids to about 900 amino acids, about 450 amino acids to about 850 amino acids, about 450 amino acids to about 800 amino acids, about 450 amino acids to about 750 amino acids, about 450 amino acids to about 700 amino acids, about 450 amino acids to about 650 amino acids, about 450 amino acids to about 600 amino acids, about 450 amino acids to about 550 amino acids, about 450 amino acids to about 500 amino acids, about 500 amino acids to about 1400 amino acids, about 500 amino acids to about 1350 amino acids, about 500 amino acids to about 1300 amino acids, about 500 amino acids to about 1250 amino acids, about 500 amino acids to about 1200 amino acids, about 500 amino acids to about 1150 amino acids, about 500 amino acids to about 1100 amino acids, about 500 amino acids to about 1050 amino acids, about 500 amino acids to about 1000 amino acids, about 500 amino acids to about 950 amino acids, about 500 amino acids to about 900 amino acids, about 500 amino acids to about 850 amino acids, about 500 amino acids to about 800 amino acids, about 500 amino acids to about 750 amino acids, about 500 amino acids to about 700 amino acids, about 500 amino acids to about 650 amino acids, about 500 amino acids to about 600 amino acids, about 500 amino acids to about 5 about 550 amino acids to about 1400 amino acids, about 550 amino acids to about 1350 amino acids, about 550 amino acids to about 1300 amino acids, about 550 amino acids to about 1250 amino acids, about 550 amino acids to about 1200 amino acids, about 550 amino acids to about 1150 amino acids, about 550 amino acids to about 1100 amino acids, about 550 amino acids to about 1050 amino acids, about 550 amino acids to about 1000 amino acids, about 550 amino acids to about 950 amino acids, about 550 amino acids to about 900 amino acids, about 550 amino acids to about 850 amino acids, about 550 amino acids to about 800 amino acids,about 550 amino acids to about 750 amino acids, about 550 amino acids to about 700 amino acids, about 550 amino acids to about 650 amino acids, about 550 amino acids to about 600 amino acids, about 600 amino acids to about 1400 amino acids, about 600 amino acids to about 1350 amino acids, about 600 amino acids to about 1300 amino acids, about 600 amino acids to about 1250 amino acids, about 600 amino acids to about 1200 amino acids, about 600 amino acids to about 1150 amino acids, about 600 amino acids to about 1100 amino acids, about 600 amino acids to about 1050 amino acids, about 600 amino acids to about 1000 amino acids, about 600 amino acids to about 1 The present invention relates to a polypeptide having an amino acid sequence of at least 200 amino acids, ... about 1100 amino acids, about 650 amino acids to about 1050 amino acids, about 650 amino acids to about 1000 amino acids, about 650 amino acids to about 950 amino acids, about 650 amino acids to about 900 amino acids, about 650 amino acids to about 850 amino acids, about 650 amino acids to about 800 amino acids, about 650 amino acids to about 750 amino acids, about 650 amino acids to about 700 amino acids, about 700 amino acids to about 1400 amino acids, about 700 amino acids to about 1350 amino acids, about 700 amino acids to about 1300 amino acids, about 700 amino acids to about 1250 amino acids, about 700 amino acids to about 1200 amino acids, about 700 amino acids to about 1150 amino acids, about 700 amino acids to about 1100 amino acids, about 700 amino acids to about 1050 amino acids, about 700 amino acids to about 1000 amino acids, about 700 amino acids to about 950 amino acids, about 700 amino acids to about 900 amino acids, about 700 amino acids to about 850 amino acids, about 700 amino acids to about 800 amino acids, about 700 amino acids to about 750 amino acids, about 750 amino acids to about 1400 amino acids, about 750 amino acids to about 1350 amino acids, about 750 amino acids to about 1300 amino acids, about 750 amino acids to about 1250 amino acids,about 750 amino acids to about 1200 amino acids, about 750 amino acids to about 1150 amino acids, about 750 amino acids to about 1100 amino acids, about 750 amino acids to about 1050 amino acids, about 750 amino acids to about 1000 amino acids, about 750 amino acids to about 950 amino acids, about 750 amino acids to about 900 amino acids, about 750 amino acids to about 850 amino acids, about 750 amino acids to about 800 amino acids, about 800 amino acids to about 1400 amino acids, about 800 amino acids to about 1350 amino acids, about 800 amino acids to about 1300 amino acids, about 800 amino acids to about 1250 amino acids, about 800 amino acids to about The present invention relates to a polypeptide having at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. The present invention relates to a polypeptide having a molecular weight of 50 amino acids, about 850 amino acids to about 1100 amino acids, about 850 amino acids to about 1050 amino acids, about 850 amino acids to about 1000 amino acids, about 850 amino acids to about 950 amino acids, about 850 amino acids to about 900 amino acids, about 900 amino acids to about 1400 amino acids, about 900 amino acids to about 1350 amino acids, about 900 amino acids to about 1300 amino acids, about 900 amino acids to about 1250 amino acids, about 900 amino acids to about 1200 amino acids, about 900 amino acids to about 1150 amino acids, about 900 amino acids to about 1100 amino acids, about 900 amino acids to about 1050 amino acids. acid, about 900 amino acids to about 1000 amino acids, about 900 amino acids to about 950 amino acids, about 950 amino acids to about 1400 amino acids, about 950 amino acids to about 1350 amino acids, about 950 amino acids to about 1300 amino acids, about 950 amino acids to about 1250 amino acids, about 950 amino acids to about 1200 amino acids, about 950 amino acids to about 1150 amino acids, about 950 amino acids to about 1100 amino acids, about 950 amino acids to about 1050 amino acids, about 950 amino acids to about 1000 amino acids, about 1000 amino acids to about 1400 amino acids, about 1000 amino acids to about 1350 amino acids,about 1000 amino acids to about 1300 amino acids, about 1000 amino acids to about 1250 amino acids, about 1000 amino acids to about 1200 amino acids, about 1000 amino acids to about 1150 amino acids, about 1000 amino acids to about 1100 amino acids, about 1000 amino acids to about 1050 amino acids, about 1050 amino acids to about 1400 amino acids, about 1050 amino acids to about 1350 amino acids, about 1050 amino acids to about about 1050 amino acids to about 1250 amino acids, about 1050 amino acids to about 1200 amino acids, about 1050 amino acids to about 1150 amino acids, about 1050 amino acids to about 1100 amino acids, about 1100 amino acids to about 1400 amino acids, about 1100 amino acids to about 1350 amino acids, about 1100 amino acids to about 1300 amino acids, about 1100 amino acids to about 1250 amino acids, about 1 The invention relates to a polypeptide having a molecular weight of 100 amino acids to about 1200 amino acids, about 1100 amino acids to about 1150 amino acids, about 1150 amino acids to about 1400 amino acids, about 1150 amino acids to about 1350 amino acids, about 1150 amino acids to about 1300 amino acids, about 1150 amino acids to about 1250 amino acids, about 1150 amino acids to about 1200 amino acids, about 1200 amino acids to about 1400 amino acids, about 1200 amino acids to about 1350 amino acids, about 1150 amino acids to about 1300 amino acids, about 1150 amino acids to about 1250 amino acids, about 1150 amino acids to about 1200 amino acids, about 1200 amino acids to about 1400 amino acids, about 1200 amino acids to about 13 50 amino acids, about 1200 amino acids to about 1300 amino acids, about 1200 amino acids to about 1250 amino acids, about 1250 amino acids to about 1400 amino acids, about 1250 amino acids to about 1350 amino acids, about 1250 amino acids to about 1300 amino acids, about 1300 amino acids to about 1400 amino acids, about 1300 amino acids to about 1350 amino acids, or about 1350 amino acids to about 1400 amino acids in total length.

[0191] For example, the chimeric polypeptide can include: LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRPWGILAESTHHKPWTATGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 44). In some cases, the chimeric polypeptide includes a β-TCP binding sequence and a linker having an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) sequence identity to SEQ ID NO: 44.

[0192] For example, the chimeric polypeptide can include: LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 45). In some cases, the chimeric polypeptide includes a β-TCP binding sequence and a linker having an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) sequence identity to SEQ ID NO: 45.

[0193] The K of any chimeric polypeptide described herein for binding to β-TCP (e.g., any type of β-TCP described herein) can be determined using a variety of different methods known in the art. D values ​​(e.g., electrophoretic mobility shift assays, filter binding assays, surface plasmon resonance, and biomolecule binding kinetics assays, etc.).

[0194] The chimeric polypeptides provided herein are useful orthopedic materials and can be used as bone void fillers and for bone reconstruction.The chimeric polypeptides described herein are osteoconductive and readily resorbable.

[0195] Linker

[0196] In some cases, the adjacent pairs (or each adjacent pairs) of two β-TCP binding sequences are directly adjacent to each other. In other cases, the adjacent pairs (or each adjacent pairs) of two β-TCP binding sequences are separated by a linker sequence. For example, the linker sequence can be 1 amino acid to about 100 amino acids, 1 amino acid to about 95 amino acids, 1 amino acid to about 90 amino acids, 1 amino acid to about 85 amino acids, 1 amino acid to about 80 amino acids, 1 amino acid to about 75 amino acids, 1 amino acid to about 70 amino acids, 1 amino acid to about 65 amino acids, 1 amino acid to about 60 amino acids, 1 amino acid to about 55 amino acids, 1 amino acid to about 50 amino acids, 1 amino acid to about 45 amino acids, 1 amino acid to about 40 amino acids, 1 amino acid to about 38 amino acids, 1 amino acid to about 36 amino acids, 1 amino acid to about 3 4 amino acids, 1 amino acid to about 32 amino acids, 1 amino acid to about 30 amino acids, 1 amino acid to about 28 amino acids, 1 amino acid to about 26 amino acids, 1 amino acid to about 24 amino acids, 1 amino acid to about 22 amino acids, 1 amino acid to about 20 amino acids, 1 amino acid to about 18 amino acids, 1 amino acid to about 16 amino acids, 1 amino acid to about 14 amino acids, 1 amino acid to about 12 amino acids, 1 amino acid to about 10 amino acids, 1 amino acid to about 8 amino acids, 1 amino acid to about 6 amino acids, 1 amino acid to about 4 amino acids, about 2 amino acids to about 100 amino acids, about 2 From about 2 amino acids to about 95 amino acids, from about 2 amino acids to about 90 amino acids, from about 2 amino acids to about 85 amino acids, from about 2 amino acids to about 80 amino acids, from about 2 amino acids to about 75 amino acids, from about 2 amino acids to about 70 amino acids, from about 2 amino acids to about 65 amino acids, from about 2 amino acids to about 60 amino acids, from about 2 amino acids to about 55 amino acids, from about 2 amino acids to about 50 amino acids, from about 2 amino acids to about 45 amino acids, from about 2 amino acids to about 40 amino acids, from about 2 amino acids to about 38 amino acids, from about 2 amino acids to about 36 amino acids, from about 2 amino acids to about 34 amino acids, from about 2 amino acids to about 36 amino acids about 32 amino acids, about 2 amino acids to about 30 amino acids, about 2 amino acids to about 28 amino acids, about 2 amino acids to about 26 amino acids, about 2 amino acids to about 24 amino acids, about 2 amino acids to about 22 amino acids, about 2 amino acids to about 20 amino acids, about 2 amino acids to about 18 amino acids, about 2 amino acids to about 16 amino acids, about 2 amino acids to about 14 amino acids, about 2 amino acids to about 12 amino acids, about 2 amino acids to about 10 amino acids, about 2 amino acids to about 8 amino acids, about 2 amino acids to about 6 amino acids, about 2 amino acids to about 4 amino acids, about 4 amino acids to about 100 amino acids,from about 4 amino acids to about 95 amino acids, from about 4 amino acids to about 90 amino acids, from about 4 amino acids to about 85 amino acids, from about 4 amino acids to about 80 amino acids, from about 4 amino acids to about 75 amino acids, from about 4 amino acids to about 70 amino acids, from about 4 amino acids to about 65 amino acids, from about 4 amino acids to about 60 amino acids, from about 4 amino acids to about 55 amino acids, from about 4 amino acids to about 50 amino acids, from about 4 amino acids to about 45 amino acids, from about 4 amino acids to about 40 amino acids, from about 4 amino acids to about 38 amino acids, from about 4 amino acids to about 36 amino acids, from about 4 amino acids to about 34 amino acids, from about 4 amino acids to about 32 amino acids, from about 4 amino acids to about about 30 amino acids, about 4 amino acids to about 28 amino acids, about 4 amino acids to about 26 amino acids, about 4 amino acids to about 24 amino acids, about 4 amino acids to about 22 amino acids, about 4 amino acids to about 20 amino acids, about 4 amino acids to about 18 amino acids, about 4 amino acids to about 16 amino acids, about 4 amino acids to about 14 amino acids, about 4 amino acids to about 12 amino acids, about 4 amino acids to about 10 amino acids, about 4 amino acids to about 8 amino acids, about 4 amino acids to about 6 amino acids, about 6 amino acids to about 100 amino acids, about 6 amino acids to about 95 amino acids, about 6 amino acids to about 90 amino acids, about 6 amino acids to about 85 amino acids, from about 6 amino acids to about 80 amino acids, from about 6 amino acids to about 75 amino acids, from about 6 amino acids to about 70 amino acids, from about 6 amino acids to about 65 amino acids, from about 6 amino acids to about 60 amino acids, from about 6 amino acids to about 55 amino acids, from about 6 amino acids to about 50 amino acids, from about 6 amino acids to about 45 amino acids, from about 6 amino acids to about 40 amino acids, from about 6 amino acids to about 38 amino acids, from about 6 amino acids to about 36 amino acids, from about 6 amino acids to about 34 amino acids, from about 6 amino acids to about 32 amino acids, from about 6 amino acids to about 30 amino acids, from about 6 amino acids to about 28 amino acids, from about 6 amino acids to about 26 amino acids, from about 6 amino acids to about about 24 amino acids, about 6 amino acids to about 22 amino acids, about 6 amino acids to about 20 amino acids, about 6 amino acids to about 18 amino acids, about 6 amino acids to about 16 amino acids, about 6 amino acids to about 14 amino acids, about 6 amino acids to about 12 amino acids, about 6 amino acids to about 10 amino acids, about 6 amino acids to about 8 amino acids, about 8 amino acids to about 100 amino acids, about 8 amino acids to about 95 amino acids, about 8 amino acids to about 90 amino acids, about 8 amino acids to about 85 amino acids, about 8 amino acids to about 80 amino acids, about 8 amino acids to about 75 amino acids, about 8 amino acids to about 70 amino acids, about 8 amino acids to about 65 amino acids,from about 8 amino acids to about 60 amino acids, from about 8 amino acids to about 55 amino acids, from about 8 amino acids to about 50 amino acids, from about 8 amino acids to about 45 amino acids, from about 8 amino acids to about 40 amino acids, from about 8 amino acids to about 38 amino acids, from about 8 amino acids to about 36 amino acids, from about 8 amino acids to about 34 amino acids, from about 8 amino acids to about 32 amino acids, from about 8 amino acids to about 30 amino acids, from about 8 amino acids to about 28 amino acids, from about 8 amino acids to about 26 amino acids, from about 8 amino acids to about 24 amino acids, from about 8 amino acids to about 22 amino acids, from about 8 amino acids to about 20 amino acids, from about 8 amino acids to about 18 amino acids, from about 8 amino acids to about 19 amino acids, from about 8 amino acids to about 21 amino acids, from about 8 amino acids to about 23 amino acids, from about 8 amino acids to about 24 amino acids, from about 8 amino acids to about 26 amino acids, from about 8 amino acids to about 24 amino acids, from about 8 amino acids to about 22 amino acids, about 16 amino acids, about 8 amino acids to about 14 amino acids, about 8 amino acids to about 12 amino acids, about 8 amino acids to about 10 amino acids, about 10 amino acids to about 100 amino acids, about 10 amino acids to about 95 amino acids, about 10 amino acids to about 90 amino acids, about 10 amino acids to about 85 amino acids, about 10 amino acids to about 80 amino acids, about 10 amino acids to about 75 amino acids, about 10 amino acids to about 70 amino acids, about 10 amino acids to about 65 amino acids, about 10 amino acids to about 60 amino acids, about 10 amino acids to about 55 amino acids, about 10 amino acids to about 50 amino acids, about 10 amino acids to about 45 amino acids, about 1 0 amino acids to about 40 amino acids, about 10 amino acids to about 38 amino acids, about 10 amino acids to about 36 amino acids, about 10 amino acids to about 34 amino acids, about 10 amino acids to about 32 amino acids, about 10 amino acids to about 30 amino acids, about 10 amino acids to about 28 amino acids, about 10 amino acids to about 26 amino acids, about 10 amino acids to about 24 amino acids, about 10 amino acids to about 22 amino acids, about 10 amino acids to about 20 amino acids, about 10 amino acids to about 18 amino acids, about 10 amino acids to about 16 amino acids, about 10 amino acids to about 14 amino acids, about 10 amino acids to about 12 amino acids, about 12 amino acids to about 12 amino acids to about 95 amino acids, about 12 amino acids to about 90 amino acids, about 12 amino acids to about 85 amino acids, about 12 amino acids to about 80 amino acids, about 12 amino acids to about 75 amino acids, about 12 amino acids to about 70 amino acids, about 12 amino acids to about 65 amino acids, about 12 amino acids to about 60 amino acids, about 12 amino acids to about 55 amino acids, about 12 amino acids to about 50 amino acids, about 12 amino acids to about 45 amino acids, about 12 amino acids to about 40 amino acids, about 12 amino acids to about 38 amino acids, about 12 amino acids to about 36 amino acids, about 12 amino acids to about 34 amino acids,from about 12 amino acids to about 32 amino acids, from about 12 amino acids to about 30 amino acids, from about 12 amino acids to about 28 amino acids, from about 12 amino acids to about 26 amino acids, from about 12 amino acids to about 24 amino acids, from about 12 amino acids to about 22 amino acids, from about 12 amino acids to about 20 amino acids, from about 12 amino acids to about 18 amino acids, from about 12 amino acids to about 16 amino acids, from about 12 amino acids to about 14 amino acids, from about 14 amino acids to about 100 amino acids, from about 14 amino acids to about 95 amino acids, from about 14 amino acids to about 90 amino acids, from about 14 amino acids to about 85 amino acids, from about 14 amino acids to about 80 amino acids, from about 14 amino acids to about about 14 amino acids to about 32 amino acids, about 14 amino acids to about 30 amino acids, about 14 amino acids to about 28 amino acids, about 14 amino acids to about 26 amino acids, about 14 amino acids to about 24 amino acids, about 14 amino acids to about 38 amino acids, about 14 amino acids to about 36 amino acids, about 14 amino acids to about 34 amino acids, about 14 amino acids to about 32 amino acids, about 14 amino acids to about 30 amino acids, about 14 amino acids to about 28 amino acids, about 14 amino acids to about 26 amino acids, about 14 amino acids to about 24 amino acids, from about 14 amino acids to about 22 amino acids, from about 14 amino acids to about 20 amino acids, from about 14 amino acids to about 18 amino acids, from about 14 amino acids to about 16 amino acids, from about 16 amino acids to about 100 amino acids, from about 16 amino acids to about 95 amino acids, from about 16 amino acids to about 90 amino acids, from about 16 amino acids to about 85 amino acids, from about 16 amino acids to about 80 amino acids, from about 16 amino acids to about 75 amino acids, from about 16 amino acids to about 70 amino acids, from about 16 amino acids to about 65 amino acids, from about 16 amino acids to about 60 amino acids, from about 16 amino acids to about 55 amino acids, from about 16 amino acids to about 50 amino acids, from about 16 amino acids to about 5 about 45 amino acids, about 16 amino acids to about 40 amino acids, about 16 amino acids to about 38 amino acids, about 16 amino acids to about 36 amino acids, about 16 amino acids to about 34 amino acids, about 16 amino acids to about 32 amino acids, about 16 amino acids to about 30 amino acids, about 16 amino acids to about 28 amino acids, about 16 amino acids to about 26 amino acids, about 16 amino acids to about 24 amino acids, about 16 amino acids to about 22 amino acids, about 16 amino acids to about 20 amino acids, about 16 amino acids to about 18 amino acids, about 18 amino acids to about 100 amino acids, about 18 amino acids to about 95 amino acids, about 18 amino acids to about 90 amino acids,from about 18 amino acids to about 85 amino acids, from about 18 amino acids to about 80 amino acids, from about 18 amino acids to about 75 amino acids, from about 18 amino acids to about 70 amino acids, from about 18 amino acids to about 65 amino acids, from about 18 amino acids to about 60 amino acids, from about 18 amino acids to about 55 amino acids, from about 18 amino acids to about 50 amino acids, from about 18 amino acids to about 45 amino acids, from about 18 amino acids to about 40 amino acids, from about 18 amino acids to about 38 amino acids, from about 18 amino acids to about 36 amino acids, from about 18 amino acids to about 34 amino acids, from about 18 amino acids to about 32 amino acids, from about 18 amino acids to about 30 amino acids, from about 18 amino acids to about 3 about 28 amino acids, about 18 amino acids to about 26 amino acids, about 18 amino acids to about 24 amino acids, about 18 amino acids to about 22 amino acids, about 18 amino acids to about 20 amino acids, about 20 amino acids to about 100 amino acids, about 20 amino acids to about 95 amino acids, about 20 amino acids to about 90 amino acids, about 20 amino acids to about 85 amino acids, about 20 amino acids to about 80 amino acids, about 20 amino acids to about 75 amino acids, about 20 amino acids to about 70 amino acids, about 20 amino acids to about 65 amino acids, about 20 amino acids to about 60 amino acids, about 20 amino acids to about 55 amino acids, about 20 amino acids to about 50 amino acids, from about 20 amino acids to about 45 amino acids, from about 20 amino acids to about 40 amino acids, from about 20 amino acids to about 38 amino acids, from about 20 amino acids to about 36 amino acids, from about 20 amino acids to about 34 amino acids, from about 20 amino acids to about 32 amino acids, from about 20 amino acids to about 30 amino acids, from about 20 amino acids to about 28 amino acids, from about 20 amino acids to about 26 amino acids, from about 20 amino acids to about 24 amino acids, from about 20 amino acids to about 22 amino acids, from about 22 amino acids to about 100 amino acids, from about 22 amino acids to about 95 amino acids, from about 22 amino acids to about 90 amino acids, from about 22 amino acids to about 85 amino acids, from about 22 amino acids to about about 22 amino acids to about 80 amino acids, about 22 amino acids to about 75 amino acids, about 22 amino acids to about 70 amino acids, about 22 amino acids to about 65 amino acids, about 22 amino acids to about 60 amino acids, about 22 amino acids to about 55 amino acids, about 22 amino acids to about 50 amino acids, about 22 amino acids to about 45 amino acids, about 22 amino acids to about 40 amino acids, about 22 amino acids to about 38 amino acids, about 22 amino acids to about 36 amino acids, about 22 amino acids to about 34 amino acids, about 22 amino acids to about 32 amino acids, about 22 amino acids to about 30 amino acids, about 22 amino acids to about 28 amino acids, about 22 amino acids to about 26 amino acids,from about 22 amino acids to about 24 amino acids, from about 24 amino acids to about 100 amino acids, from about 24 amino acids to about 95 amino acids, from about 24 amino acids to about 90 amino acids, from about 24 amino acids to about 85 amino acids, from about 24 amino acids to about 80 amino acids, from about 24 amino acids to about 75 amino acids, from about 24 amino acids to about 70 amino acids, from about 24 amino acids to about 65 amino acids, from about 24 amino acids to about 60 amino acids, from about 24 amino acids to about 55 amino acids, from about 24 amino acids to about 50 amino acids, from about 24 amino acids to about 45 amino acids, from about 24 amino acids to about 40 amino acids, from about 24 amino acids to about 38 amino acids, from about 24 amino acids to about 5 about 24 amino acids to about 34 amino acids, about 24 amino acids to about 32 amino acids, about 24 amino acids to about 30 amino acids, about 24 amino acids to about 28 amino acids, about 24 amino acids to about 26 amino acids, about 26 amino acids to about 100 amino acids, about 26 amino acids to about 95 amino acids, about 26 amino acids to about 90 amino acids, about 26 amino acids to about 85 amino acids, about 26 amino acids to about 80 amino acids, about 26 amino acids to about 75 amino acids, about 26 amino acids to about 70 amino acids, about 26 amino acids to about 65 amino acids, about 26 amino acids to about 60 amino acids, about 26 amino acids to about 55 amino acids, from about 26 amino acids to about 50 amino acids, from about 26 amino acids to about 45 amino acids, from about 26 amino acids to about 40 amino acids, from about 26 amino acids to about 38 amino acids, from about 26 amino acids to about 36 amino acids, from about 26 amino acids to about 34 amino acids, from about 26 amino acids to about 32 amino acids, from about 26 amino acids to about 30 amino acids, from about 26 amino acids to about 28 amino acids, from about 28 amino acids to about 100 amino acids, from about 28 amino acids to about 95 amino acids, from about 28 amino acids to about 90 amino acids, from about 28 amino acids to about 85 amino acids, from about 28 amino acids to about 80 amino acids, from about 28 amino acids to about 75 amino acids, from about 28 amino acids to about 10 about 70 amino acids, about 28 amino acids to about 65 amino acids, about 28 amino acids to about 60 amino acids, about 28 amino acids to about 55 amino acids, about 28 amino acids to about 50 amino acids, about 28 amino acids to about 45 amino acids, about 28 amino acids to about 40 amino acids, about 28 amino acids to about 38 amino acids, about 28 amino acids to about 36 amino acids, about 28 amino acids to about 34 amino acids, about 28 amino acids to about 32 amino acids, about 28 amino acids to about 30 amino acids, about 30 amino acids to about 100 amino acids, about 30 amino acids to about 95 amino acids, about 30 amino acids to about 90 amino acids, about 30 amino acids to about 85 amino acids,from about 30 amino acids to about 80 amino acids, from about 30 amino acids to about 75 amino acids, from about 30 amino acids to about 70 amino acids, from about 30 amino acids to about 65 amino acids, from about 30 amino acids to about 60 amino acids, from about 30 amino acids to about 55 amino acids, from about 30 amino acids to about 50 amino acids, from about 30 amino acids to about 45 amino acids, from about 30 amino acids to about 40 amino acids, from about 30 amino acids to about 38 amino acids, from about 30 amino acids to about 36 amino acids, from about 30 amino acids to about 34 amino acids, from about 30 amino acids to about 32 amino acids, from about 32 amino acids to about 100 amino acids, from about 32 amino acids to about 95 amino acids, from about 32 amino acids to about 106 amino acids, from about 32 amino acids to about 114 amino acids, from about 32 amino acids to about 129 amino acids, from about 32 amino acids to about 130 amino acids, from about 32 amino acids to about 145 amino acids, from about 32 amino acids to about 150 amino acids, from about 32 amino acids to about 160 amino acids, from about 32 amino acids to about 170 amino acids, from about 32 amino acids to about 180 amino acids, from about 32 amino acids to about 190 amino acids, from about 32 amino acids to about 195 amino acids, from about 32 amino acids to about 180 amino acids, from about about 90 amino acids, about 32 amino acids to about 85 amino acids, about 32 amino acids to about 80 amino acids, about 32 amino acids to about 75 amino acids, about 32 amino acids to about 70 amino acids, about 32 amino acids to about 65 amino acids, about 32 amino acids to about 60 amino acids, about 32 amino acids to about 55 amino acids, about 32 amino acids to about 50 amino acids, about 32 amino acids to about 45 amino acids, about 32 amino acids to about 40 amino acids, about 32 amino acids to about 38 amino acids, about 32 amino acids to about 36 amino acids, about 32 amino acids to about 34 amino acids, about 34 amino acids to about 100 amino acids, about 34 amino acids to about 95 amino acids, from about 34 amino acids to about 90 amino acids, from about 34 amino acids to about 85 amino acids, from about 34 amino acids to about 80 amino acids, from about 34 amino acids to about 75 amino acids, from about 34 amino acids to about 70 amino acids, from about 34 amino acids to about 65 amino acids, from about 34 amino acids to about 60 amino acids, from about 34 amino acids to about 55 amino acids, from about 34 amino acids to about 50 amino acids, from about 34 amino acids to about 45 amino acids, from about 34 amino acids to about 40 amino acids, from about 34 amino acids to about 38 amino acids, from about 34 amino acids to about 36 amino acids, from about 36 amino acids to about 100 amino acids, from about 36 amino acids to about 95 amino acids, from about 36 amino acids to about 10 ... about 90 amino acids, about 36 amino acids to about 85 amino acids, about 36 amino acids to about 80 amino acids, about 36 amino acids to about 75 amino acids, about 36 amino acids to about 70 amino acids, about 36 amino acids to about 65 amino acids, about 36 amino acids to about 60 amino acids, about 36 amino acids to about 55 amino acids, about 36 amino acids to about 50 amino acids, about 36 amino acids to about 45 amino acids, about 36 amino acids to about 40 amino acids, about 36 amino acids to about 38 amino acids, about 38 amino acids to about 100 amino acids, about 38 amino acids to about 95 amino acids, about 38 amino acids to about 90 amino acids, about 38 amino acids to about 85 amino acids,from about 38 amino acids to about 80 amino acids, from about 38 amino acids to about 75 amino acids, from about 38 amino acids to about 70 amino acids, from about 38 amino acids to about 65 amino acids, from about 38 amino acids to about 60 amino acids, from about 38 amino acids to about 55 amino acids, from about 38 amino acids to about 50 amino acids, from about 38 amino acids to about 45 amino acids, from about 38 amino acids to about 40 amino acids, from about 40 amino acids to about 100 amino acids, from about 40 amino acids to about 95 amino acids, from about 40 amino acids to about 90 amino acids, from about 40 amino acids to about 85 amino acids, from about 40 amino acids to about 80 amino acids, from about 40 amino acids to about 75 amino acids, from about 40 amino acids to about 50 about 70 amino acids, about 40 amino acids to about 65 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 55 amino acids, about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 45 amino acids to about 100 amino acids, about 45 amino acids to about 95 amino acids, about 45 amino acids to about 90 amino acids, about 45 amino acids to about 85 amino acids, about 45 amino acids to about 80 amino acids, about 45 amino acids to about 75 amino acids, about 45 amino acids to about 70 amino acids, about 45 amino acids to about 65 amino acids, about 45 amino acids to about 60 amino acids, about 45 amino acids to about 55 amino acids, about from about 45 amino acids to about 50 amino acids, from about 50 amino acids to about 100 amino acids, from about 50 amino acids to about 95 amino acids, from about 50 amino acids to about 90 amino acids, from about 50 amino acids to about 85 amino acids, from about 50 amino acids to about 80 amino acids, from about 50 amino acids to about 75 amino acids, from about 50 amino acids to about 70 amino acids, from about 50 amino acids to about 65 amino acids, from about 50 amino acids to about 60 amino acids, from about 50 amino acids to about 55 amino acids, from about 55 amino acids to about 100 amino acids, from about 55 amino acids to about 95 amino acids, from about 55 amino acids to about 90 amino acids, from about 55 amino acids to about 85 amino acids, from about 55 amino acids to about about 80 amino acids, about 55 amino acids to about 75 amino acids, about 55 amino acids to about 70 amino acids, about 55 amino acids to about 65 amino acids, about 55 amino acids to about 60 amino acids, about 60 amino acids to about 100 amino acids, about 60 amino acids to about 95 amino acids, about 60 amino acids to about 90 amino acids, about 60 amino acids to about 85 amino acids, about 60 amino acids to about 80 amino acids, about 60 amino acids to about 75 amino acids, about 60 amino acids to about 70 amino acids, about 60 amino acids to about 65 amino acids, about 65 amino acids to about 100 amino acids, about 65 amino acids to about 95 amino acids, about 65 amino acids to about 90 amino acids,from about 65 amino acids to about 85 amino acids, from about 65 amino acids to about 80 amino acids, from about 65 amino acids to about 75 amino acids, from about 65 amino acids to about 70 amino acids, from about 70 amino acids to about 100 amino acids, from about 70 amino acids to about 95 amino acids, from about 70 amino acids to about 90 amino acids, from about 70 amino acids to about 85 amino acids, from about 70 amino acids to about 80 amino acids, from about 70 amino acids to about 75 amino acids, from about 75 amino acids to about 100 amino acids, from about 75 amino acids to about 95 amino acids, from about 75 amino acids to about 90 amino acids, about 75 amino acids to about 85 amino acids, about 75 amino acids to about 80 amino acids, about 80 amino acids to about 100 amino acids, about 80 amino acids to about 95 amino acids, about 80 amino acids to about 90 amino acids, about 80 amino acids to about 85 amino acids, about 85 amino acids to about 100 amino acids, about 85 amino acids to about 95 amino acids, about 85 amino acids to about 90 amino acids, about 90 amino acids to about 100 amino acids, about 90 amino acids to about 95 amino acids, or about 95 amino acids to about 100 amino acids.

[0197] For example, the linker sequence can comprise or consist of TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33) or GSGATG (SEQ ID NO: 34). In some cases, the chimeric polypeptide includes a linker having an amino acid sequence that has at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) sequence identity to SEQ ID NO: 33. For example, the linker sequence can comprise or consist of the sequence of GSGS, GSGSGS, SGSG, SGSGSG, GSSG, SS, or GGGS.

[0198] Growth Factors

[0199] The β-TCP-binding sequences described herein can be tethered to a mammalian growth factor. In some cases, a linker sequence (eg, any linker sequence described herein or known in the art) is positioned between the β-TCP-binding sequence and the mammalian growth factor.

[0200] Mammalian growth factors can be osteoinductive molecules that are able to initiate and enhance the bone repair process. Bone morphogenetic proteins (BMPs) represent a unique subset of the transforming growth factor-β family. Many of these BMPs (BMP-2, BMP-7, and BMP-14) increase the rate of bone healing in defects and nonunions.

[0201] Non-limiting examples of mammalian growth factors are described herein. In some cases,The mammalian growth factor is selected from the group consisting of epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF-1), fibroblast growth factor (FGF), fibroblast growth factor 2 (FGF2), fibroblast growth factor 18 (FGF18), transforming growth factor α (TGF-α), transforming growth factor β (TGF-β), transforming growth factor β1 (TGF-β1), transforming growth factor β3 (TGF-β3), osteogenic protein 1 (OP-1), osteogenic protein 2 (OP-2), osteogenic protein 3 (OP-3), bone morphogenetic protein 2 (BMP-2), bone morphogenetic protein 3 (BMP-3), bone morphogenetic protein 4 ( BMP-4), bone morphogenetic protein 5 (BMP-5), bone morphogenetic protein 6 (BMP-6), bone morphogenetic protein 7 (BMP-7), bone morphogenetic protein (BMP-9), bone morphogenetic protein 10 (BMP-10), bone morphogenetic protein 11 (BMP-11), bone morphogenetic protein 12 (BMP-12), bone morphogenetic protein protein 13 (BMP-13), bone morphogenetic protein 15 (BMP-15), dentin phosphoprotein (DPP), plant-related growth factor (Vgr), growth differentiation factor 1 (GDF-1), growth differentiation factor 3 (GDF-3), growth differentiation factor 5 (GDF-5), growth differentiation factor 6 (GDF-6), Growth differentiation factor 7 (GDF-7), growth differentiation factor 8 (GDF8), growth differentiation factor 11 (GDF11), growth differentiation factor 15 (GDF15), vascular endothelial growth factor (VEGF), hyaluronan binding protein (HABP) and collagen binding protein (CBP), fibroblast growth factor 18 (FGF-18), keratinocyte growth factor (KGF), tumor necrosis factor α (TNFα), tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), wnt family member 1 (WNT1), wnt family member 2 (WNT2), wnt family member 2B (WNT2B), wnt family member 3 (WNT3), wnt family Wnt family member 3A (WNT3A), wnt family member 4 (WNT4), wnt family member 5A (WNT5A), wnt family member 5B (WNT5B), wnt family member 6 (WNT6), wnt family member 7A (WNT7A), wnt family member 7B (WNT7B), wnt family member 8A (WNT8A), wnt family member 8B (WNT8B), wnt family member 9A (WNT9A), wnt family member 9B (WNT9B), wnt family member 10A (WNT10A), wnt family member 10B (WNT10B), wnt family member 11 (WNT11) and wnt family member 16 (WNT16).

[0202] For example, the mammalian growth factor can be a human growth factor. Non-limiting examples of human growth factors are provided in Table B.

[0203] Table B. Exemplary Human Growth Factors

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221]

[0222]

[0223]

[0224]

[0225] In some embodiments, the mammalian growth factor is a secreted human growth factor. In some embodiments, the mammalian growth factor is an active fragment having osteogenic activity (e.g., a fragment of a secreted human growth factor (e.g., any exemplary growth factor described herein)). Non-limiting in vitro assays for determining whether a fragment of a secreted human growth factor (e.g., any exemplary growth factor described herein) has osteogenic activity are described in Kim et al., Amino Acids 42: 1455-1465, 2012; Lee et al., ACS Med. Chem. Lett. 2(3): 248-251, 2011; and Wang et al., Genetics Mol. Res. 13(2): 4456-4465, 2014.

[0226] In some embodiments, the fragment of the secreted human growth factor having osteogenic activity can remove 1 amino acid to about 40 amino acids from the N-terminus of the secreted human growth factor (e.g., 1 amino acid to about 38 amino acids, 1 amino acid to about 36 amino acids, 1 amino acid to about 34 amino acids, 1 amino acid to about 32 amino acids, 1 amino acid to about 30 amino acids, 1 amino acid to about 28 amino acids, 1 amino acid to about 26 amino acids, 1 amino acid to about 24 amino acids, 1 amino acid to about 22 amino acids, 1 amino acid to about 20 amino acids, 1 amino acid to about 18 amino acids, 1 amino acid to about 16 amino acids, 1 amino acid to about 14 amino acids). The invention also provides for removing 1 amino acid to about 12 amino acids, 1 amino acid to about 10 amino acids, 1 amino acid to about 8 amino acids, 1 amino acid to about 6 amino acids, 1 amino acid to about 4 amino acids, or about 1 amino acid to about 3 amino acids) and / or removing 1 amino acid to about 20 amino acids (e.g., 1 amino acid to about 18 amino acids, 1 amino acid to about 16 amino acids, 1 amino acid to about 14 amino acids, 1 amino acid to about 12 amino acids, 1 amino acid to about 10 amino acids, 1 amino acid to about 8 amino acids, 1 amino acid to about 6 amino acids, 1 amino acid to about 4 amino acids, or about 1 amino acid to about 3 amino acids) from the C-terminus of the secreted human growth factor.

[0227] In some embodiments, the mammalian growth factor can have a sequence that is at least 70% identical (e.g., at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical) to any sequence in Table B or any secreted human growth factor and has osteogenic activity. As will be appreciated by those skilled in the art, amino acids in mammalian growth factors that are conserved between different species may be important for osteogenic activity and should not be mutated, whereas amino acids in mammalian growth factors that are not conserved between different species may be less important for osteogenic activity and may be mutated.

[0228] β-TCP binding peptide

[0229] Also provided herein are beta-tricalcium phosphate (beta-TCP)-binding polypeptides comprising at least one (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty) selected from the following sequences: VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13); NO:13), GSSGESTHHKPST (SEQ ID NO:14), VGADSTHHRPVT (SEQ ID NO:15), GAADTTHHRPVT (SEQ ID NO:16), AGADTTHHRPVT (SEQ ID NO:17), GGADTTHHRPAT (SEQ ID NO:18) and GGADTTHHRPGT (SEQ ID NO:19).

[0230] Also provided herein are β-TCP binding polypeptides comprising the amino acid sequence of Formula I:

[0231] A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein:

[0232] A0 is V, L, I, G, S, T, or A;

[0233] B0 is I, L, V, Q, T, S, G, or A;

[0234] C0 is G, A, V or S;

[0235] D0 is E, D, L or G;

[0236] E0 is S, T, P, T, E, or D;

[0237] F0 is T or S;

[0238] G0 is H, T or S;

[0239] H0 is H or T;

[0240] I0 is R, S, K, P or H;

[0241] J0 is P, S, R or K;

[0242] K0 is W, F, S, P, V, A, or G; and

[0243] L0 does not exist or is S, T, G (or A);

[0244] Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0245] In some embodiments, any β-TCP binding polypeptide described herein can include two or more (e.g., three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, or twenty or more; or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22) any of SEQ ID NOs: 2-19 and sequences of Formula I (excluding SEQ ID NO: 1). In some embodiments, each adjacent pair of sequences of Formula I (excluding SEQ ID NO: 1) and SEQ ID NOs: 2-19 is directly adjacent to each other. In some embodiments, each adjacent pair of a sequence of Formula I (excluding SEQ ID NO: 1) and any of SEQ ID NOs: 2-19 is separated by a linker sequence.

[0246] In some embodiments, the chimeric polypeptide can include at least two (e.g., at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten; or 2, 3, 4, 5, 6, 7, 8, 9, or 10) different sequences selected from SEQ ID NOs: 2-19 and a sequence of Formula I (excluding SEQ ID NO: 1).

[0247] In some embodiments, the β-TCP binding polypeptide includes LLADTTHHRPWT (SEQ ID NO: 1) or GQVLPTTTPSSP (SEQ ID NO: 19). In some embodiments, the β-TCP binding polypeptide includes LLADTTHHRPWT (SEQ ID NO: 1) and VIGESTHHRPWS (SEQ ID NO: 2). In some embodiments, the β-TCP binding polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWS (SEQ ID NO: 20).

[0248] In some embodiments, the β-TCP binding polypeptide includes LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), and IIGESSHHKPFT (SEQ ID NO: 6). In some embodiments, the β-TCP binding polypeptide includes the sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFT (SEQ ID NO: 21).

[0249] In some embodiments, the β-TCP binding polypeptide comprises LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7) and ILAESTHHKPWT (SEQ ID NO: 4). In some embodiments, the β-TCP binding polypeptide comprises the following sequence: LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRPW GILAESTHHKPWT (SEQ ID NO: 22).

[0250] In some embodiments, the β-TCP binding polypeptide includes LLADTTHHRPWT (SEQ ID NO: 1) and ILAESTHHKPWT (SEQ ID NO: 4). In some embodiments, the β-TCP binding polypeptide includes at least one copy of the sequence LLADTTHHRPWTILAESTHHKPWT (SEQ ID NO: 23). In some embodiments, the β-TCP binding polypeptide includes the sequence of LLADTTHHRPWTILAESTHHKPWTLLADTTHHRPWTILAESTHHKP WTLLADTTHHRPWT (SEQ ID NO: 24).

[0251] In some embodiments, the β-TCP binding polypeptide comprises LLADTTHHRPWT (SEQ ID NO: 1) and GLGDTTHHRPWG (SEQ ID NO: 7). In some embodiments, the β-TCP binding polypeptide comprises at least one copy of the sequence of LLADTTHHRPWTGLGDTTHHRPWG (SEQ ID NO: 25). In some embodiments, the β-TCP binding polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 26). In some embodiments, the β-TCP binding polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 27). In some embodiments, the β-TCP binding polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 28).

[0252] In some embodiments, the β-TCP binding polypeptide comprises STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), and GSSGESTHHKPST (SEQ ID NO: 14). In some embodiments, the β-TCP binding polypeptide comprises the sequence of STADTSHHRPSTSGGESTHHRPSTSGGESSHHKPSTGSGDSSHHRPSG SSGESTHHKPST (SEQ ID NO: 29).

[0253] In some embodiments, the β-TCP binding polypeptide includes VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19). In some embodiments, the β-TCP binding polypeptide includes the sequence of VGADSTHHRPVTGAADTTHHRPVTAGADTTHHRPVTGGADTTHHR PATGGADTTHHRPGT (SEQ ID NO: 30).

[0254] In some embodiments, the β-TCP binding polypeptide comprises STADTSHHRPS (SEQ ID NO: 10), LLADTTHHRPWT (SEQ ID NO: 1), TSGGESTHHRPS (SEQ ID NO: 11), VGADSTHHRPVT (SEQ ID NO: 15), TSGGESSHHKPS (SEQ ID NO: 12), GAADTTHHRPVT (SEQ ID NO: 16), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPS (SEQ ID NO: 14) and TGGADTTHHRPAT (SEQ ID NO: 18). In some embodiments, the β-TCP binding polypeptide comprises the following sequence: STADTSHHRPSLLADTTHHRPWTTSGGESTHHRPSVGADSTHHRPVT TSGGESSHHKPSGAADTTHHRPVTTGSGDSSHHRPSGSSGESTHHKPSSTGGADTTHHRPAT (SEQ ID NO: 31).

[0255] In some embodiments, the β-TCP binding polypeptide can include two or more (e.g., three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, or twenty or more; or 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) copies of any one of SEQ ID NOs: 2-19 or Formula I (excluding SEQ ID NO: 1).

[0256] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 1 (but not including SEQ ID NO: 1).

[0257] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:2.

[0258] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:3.

[0259] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:4.

[0260] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:5.

[0261] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:6.

[0262] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:7.

[0263] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:8.

[0264] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:9.

[0265] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 10.

[0266] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 11.

[0267] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 12.

[0268] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 13.

[0269] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:14.

[0270] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO:15.

[0271] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 16.

[0272] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 17.

[0273] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 18.

[0274] In some embodiments, the β-TCP binding polypeptide can include an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO: 19.

[0275] In some embodiments, the β-TCP binding polypeptide further comprises a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) between adjacent pairs of the sequence of Formula I (excluding SEQ ID NO: 1) and any SEQ ID NO: 2-19. In some embodiments, the linker sequence may comprise the sequence of TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33). In some embodiments, the linker sequence consists of TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33). In some embodiments, the linker sequence comprises the sequence of GSGATG (SEQ ID NO: 34). In some embodiments, the linker sequence consists of GSGATG (SEQ ID NO: 34).

[0276] In some embodiments, where the β-TCP binding polypeptide includes two or more of any one of SEQ ID NOs: 2-19 and a sequence of Formula I (excluding SEQ ID NO: 1), a linker sequence (e.g., any exemplary linker sequence described herein or known in the art) may be between any pair (e.g., all pairs) of adjacent sequences selected from SEQ ID NOs: 2-19 and a sequence of Formula I (excluding SEQ ID NO: 1).

[0277] In some embodiments, the β-TCP binding polypeptide may include a signal sequence at its N-terminus. In some embodiments, the β-TCP binding polypeptide may also include a tag sequence (e.g., a poly-His tag, chitin binding protein (CBP), maltose binding protein (MBP), a streptococcal tag, glutathione-S-transferase (GST), thioredoxin, or an Fc region). Additional examples of tags are known in the art.

[0278] The β-TCP binding polypeptides described herein can be isolated with a dissociation equilibrium constant (K) of about 1 pM to about 100 μM (or any subrange of this range described herein). d ) (e.g., as measured using SPR in phosphate-buffered saline) binding to β-TCP (e.g., any type of β-TCP described herein).

[0279] In some embodiments, the total length of the β-TCP binding polypeptides described herein is from about 11 amino acids to about 1400 amino acids, from about 11 amino acids to about 1350 amino acids, from about 11 amino acids to about 1300 amino acids, from about 11 amino acids to about 1250 amino acids, from about 11 amino acids to about 1200 amino acids, from about 11 amino acids to about 1150 amino acids, from about 11 amino acids to about 1100 amino acids, from about 11 amino acids to about 1050 amino acids, from about 11 amino acids to about 1000 amino acids, from about 11 amino acids to about 950 amino acids, from about 11 amino acids to about 900 amino acids, from about 11 amino acids to about 850 amino acids, from about 11 amino acids to about 1 11 amino acids to about 800 amino acids, about 11 amino acids to about 750 amino acids, about 11 amino acids to about 700 amino acids, about 11 amino acids to about 650 amino acids, about 11 amino acids to about 600 amino acids, about 11 amino acids to about 550 amino acids, about 11 amino acids to about 500 amino acids, about 11 amino acids to about 450 amino acids, about 11 amino acids to about 400 amino acids, about 11 amino acids to about 350 amino acids, about 11 amino acids to about 300 amino acids, about 11 amino acids to about 250 amino acids, about 11 amino acids to about 200 amino acids, about 11 amino acids to about 150 amino acids, about 11 amino acids to about 100 amino acids, about 11 amino acids to about 90 amino acids, about 11 amino acids to about 80 amino acids, about 11 amino acids to about 70 amino acids, about 11 amino acids to about 60 amino acids, about 11 amino acids to about 50 amino acids, about 11 amino acids to about 45 amino acids, about 11 amino acids to about 40 amino acids, about 11 amino acids to about 35 amino acids, about 11 amino acids to about 30 amino acids, about 11 amino acids to about 25 amino acids, about 11 amino acids to about 20 amino acids, about 11 amino acids to about 18 amino acids, about 11 amino acids to about 16 amino acids, about 11 amino acids to about 14 amino acids, about 12 amino acids to about 135 amino acids, about 12 amino acids to about 1400 amino acids. 0 amino acids, about 12 amino acids to about 1300 amino acids, about 12 amino acids to about 1250 amino acids, about 12 amino acids to about 1200 amino acids, about 12 amino acids to about 1150 amino acids, about 12 amino acids to about 1100 amino acids, about 12 amino acids to about 1050 amino acids, about 12 amino acids to about 1000 amino acids, about 12 amino acids to about 950 amino acids, about 12 amino acids to about 900 amino acids, about 12 amino acids to about 850 amino acids, about 12 amino acids to about 800 amino acids, about 12 amino acids to about 750 amino acids, about 12 amino acids to about 700 amino acids, about 12 amino acids to about 650 amino acids,from about 12 amino acids to about 600 amino acids, from about 12 amino acids to about 550 amino acids, from about 12 amino acids to about 500 amino acids, from about 12 amino acids to about 450 amino acids, from about 12 amino acids to about 400 amino acids, from about 12 amino acids to about 350 amino acids, from about 12 amino acids to about 300 amino acids, from about 12 amino acids to about 250 amino acids, from about 12 amino acids to about 200 amino acids, from about 12 amino acids to about 150 amino acids, from about 12 amino acids to about 100 amino acids, from about 12 amino acids to about 90 amino acids, from about 12 amino acids to about 80 amino acids, from about 12 amino acids to about 70 amino acids, from about 12 amino acids to about 60 amino acids, about 12 amino acids to about 50 amino acids, about 12 amino acids to about 45 amino acids, about 12 amino acids to about 40 amino acids, about 12 amino acids to about 35 amino acids, about 12 amino acids to about 30 amino acids, about 12 amino acids to about 25 amino acids, about 12 amino acids to about 20 amino acids, about 12 amino acids to about 18 amino acids, about 12 amino acids to about 16 amino acids, about 12 amino acids to about 14 amino acids, about 14 amino acids to about 1400 amino acids, about 14 amino acids to about 1350 amino acids, about 14 amino acids to about 1300 amino acids, about 14 amino acids to about 1250 amino acids, about 14 amino acids to about 120 0 amino acids, about 14 amino acids to about 1150 amino acids, about 14 amino acids to about 1100 amino acids, about 14 amino acids to about 1050 amino acids, about 14 amino acids to about 1000 amino acids, about 14 amino acids to about 950 amino acids, about 14 amino acids to about 900 amino acids, about 14 amino acids to about 850 amino acids, about 14 amino acids to about 800 amino acids, about 14 amino acids to about 750 amino acids, about 14 amino acids to about 700 amino acids, about 14 amino acids to about 650 amino acids, about 14 amino acids to about 600 amino acids, about 14 amino acids to about 550 amino acids, about 14 amino acids to about 500 amino acids, from about 14 amino acids to about 450 amino acids, from about 14 amino acids to about 400 amino acids, from about 14 amino acids to about 350 amino acids, from about 14 amino acids to about 300 amino acids, from about 14 amino acids to about 250 amino acids, from about 14 amino acids to about 200 amino acids, from about 14 amino acids to about 150 amino acids, from about 14 amino acids to about 100 amino acids, from about 14 amino acids to about 90 amino acids, from about 14 amino acids to about 80 amino acids, from about 14 amino acids to about 70 amino acids, from about 14 amino acids to about 60 amino acids, from about 14 amino acids to about 50 amino acids, from about 14 amino acids to about 45 amino acids, from about 14 amino acids to about 40 amino acids,from about 14 amino acids to about 35 amino acids, from about 14 amino acids to about 30 amino acids, from about 14 amino acids to about 25 amino acids, from about 14 amino acids to about 20 amino acids, from about 14 amino acids to about 18 amino acids, from about 14 amino acids to about 16 amino acids, from about 16 amino acids to about 1400 amino acids, from about 16 amino acids to about 1350 amino acids, from about 16 amino acids to about 1300 amino acids, from about 16 amino acids to about 1250 amino acids, from about 16 amino acids to about 1200 amino acids, from about 16 amino acids to about 1150 amino acids, from about 16 amino acids to about 1100 amino acids, from about 16 amino acids to about 1050 amino acids, from about 16 amino acids to about 16 From about 16 amino acids to about 1000 amino acids, from about 16 amino acids to about 950 amino acids, from about 16 amino acids to about 900 amino acids, from about 16 amino acids to about 850 amino acids, from about 16 amino acids to about 800 amino acids, from about 16 amino acids to about 750 amino acids, from about 16 amino acids to about 700 amino acids, from about 16 amino acids to about 650 amino acids, from about 16 amino acids to about 600 amino acids, from about 16 amino acids to about 550 amino acids, from about 16 amino acids to about 500 amino acids, from about 16 amino acids to about 450 amino acids, from about 16 amino acids to about 400 amino acids, from about 16 amino acids to about 350 amino acids, from about 16 amino acids to about 300 amino acids, about 16 amino acids to about 250 amino acids, about 16 amino acids to about 200 amino acids, about 16 amino acids to about 150 amino acids, about 16 amino acids to about 100 amino acids, about 16 amino acids to about 90 amino acids, about 16 amino acids to about 80 amino acids, about 16 amino acids to about 70 amino acids, about 16 amino acids to about 60 amino acids, about 16 amino acids to about 50 amino acids, about 16 amino acids to about 45 amino acids, about 16 amino acids to about 40 amino acids, about 16 amino acids to about 35 amino acids, about 16 amino acids to about 30 amino acids, about 16 amino acids to about 25 amino acids, about 16 amino acids to about 20 amino acids , about 16 amino acids to about 18 amino acids, about 18 amino acids to about 1400 amino acids, about 18 amino acids to about 1350 amino acids, about 18 amino acids to about 1300 amino acids, about 18 amino acids to about 1250 amino acids, about 18 amino acids to about 1200 amino acids, about 18 amino acids to about 1150 amino acids, about 18 amino acids to about 1100 amino acids, about 18 amino acids to about 1050 amino acids, about 18 amino acids to about 1000 amino acids, about 18 amino acids to about 950 amino acids, about 18 amino acids to about 900 amino acids, about 18 amino acids to about 850 amino acids, about 18 amino acids to about 800 amino acids,from about 18 amino acids to about 750 amino acids, from about 18 amino acids to about 700 amino acids, from about 18 amino acids to about 650 amino acids, from about 18 amino acids to about 600 amino acids, from about 18 amino acids to about 550 amino acids, from about 18 amino acids to about 500 amino acids, from about 18 amino acids to about 450 amino acids, from about 18 amino acids to about 400 amino acids, from about 18 amino acids to about 350 amino acids, from about 18 amino acids to about 300 amino acids, from about 18 amino acids to about 250 amino acids, from about 18 amino acids to about 200 amino acids, from about 18 amino acids to about 150 amino acids, from about 18 amino acids to about 100 amino acids, from about 18 amino acids to about 9 0 amino acids, about 18 amino acids to about 80 amino acids, about 18 amino acids to about 70 amino acids, about 18 amino acids to about 60 amino acids, about 18 amino acids to about 50 amino acids, about 18 amino acids to about 45 amino acids, about 18 amino acids to about 40 amino acids, about 18 amino acids to about 35 amino acids, about 18 amino acids to about 30 amino acids, about 18 amino acids to about 25 amino acids, about 18 amino acids to about 20 amino acids, about 20 amino acids to about 1400 amino acids, about 20 amino acids to about 1350 amino acids, about 20 amino acids to about 1250 amino acids, about 20 amino acids to about 12 00 amino acids, about 20 amino acids to about 1150 amino acids, about 20 amino acids to about 1100 amino acids, about 20 amino acids to about 1050 amino acids, about 20 amino acids to about 1000 amino acids, about 20 amino acids to about 950 amino acids, about 20 amino acids to about 900 amino acids, about 20 amino acids to about 850 amino acids, about 20 amino acids to about 800 amino acids, about 20 amino acids to about 750 amino acids, about 20 amino acids to about 700 amino acids, about 20 amino acids to about 650 amino acids, about 20 amino acids to about 600 amino acids, about 20 amino acids to about 550 amino acids, about 20 amino acids to about 500 amino acids , about 20 amino acids to about 450 amino acids, about 20 amino acids to about 400 amino acids, about 20 amino acids to about 350 amino acids, about 20 amino acids to about 300 amino acids, about 20 amino acids to about 250 amino acids, about 20 amino acids to about 200 amino acids, about 20 amino acids to about 150 amino acids, about 20 amino acids to about 100 amino acids, about 20 amino acids to about 90 amino acids, about 20 amino acids to about 80 amino acids, about 20 amino acids to about 70 amino acids, about 20 amino acids to about 60 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 45 amino acids, about 20 amino acids to about 40 amino acids,about 20 amino acids to about 35 amino acids, about 20 amino acids to about 30 amino acids, about 20 amino acids to about 25 amino acids, about 25 amino acids to about 1400 amino acids, about 25 amino acids to about 1350 amino acids, about 25 amino acids to about 1300 amino acids, about 25 amino acids to about 1250 amino acids, about 25 amino acids to about 1200 amino acids, about 25 amino acids to about 1150 amino acids, about 25 amino acids to about 1100 amino acids, about 25 amino acids to about 1050 amino acids, about 25 amino acids to about 1000 amino acids, about 25 amino acids to about 950 amino acids, about 25 amino acids to about 900 amino acids, about 25 In some embodiments, the present invention relates to a polypeptide having at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. about 25 amino acids to about 100 amino acids, about 25 amino acids to about 90 amino acids, about 25 amino acids to about 80 amino acids, about 25 amino acids to about 70 amino acids, about 25 amino acids to about 60 amino acids, about 25 amino acids to about 50 amino acids, about 25 amino acids to about 45 amino acids, about 25 amino acids to about 40 amino acids, about 25 amino acids to about 35 amino acids, about 25 amino acids to about 30 amino acids, about 30 amino acids to about 1400 amino acids, about 30 amino acids to about 1350 amino acids, about 30 amino acids to about 1300 amino acids, about 30 amino acids to about 1250 amino acids, about 30 amino acids to about 120 0 amino acids, about 30 amino acids to about 1150 amino acids, about 30 amino acids to about 1100 amino acids, about 30 amino acids to about 1050 amino acids, about 30 amino acids to about 1000 amino acids, about 30 amino acids to about 950 amino acids, about 30 amino acids to about 900 amino acids, about 30 amino acids to about 850 amino acids, about 30 amino acids to about 800 amino acids, about 30 amino acids to about 750 amino acids, about 30 amino acids to about 700 amino acids, about 30 amino acids to about 650 amino acids, about 30 amino acids to about 600 amino acids, about 30 amino acids to about 550 amino acids, about 30 amino acids to about 500 amino acids,about 30 amino acids to about 450 amino acids, about 30 amino acids to about 400 amino acids, about 30 amino acids to about 350 amino acids, about 30 amino acids to about 300 amino acids, about 30 amino acids to about 250 amino acids, about 30 amino acids to about 200 amino acids, about 30 amino acids to about 150 amino acids, about 30 amino acids to about 100 amino acids, about 30 amino acids to about 90 amino acids, about 30 amino acids to about 80 amino acids, about 30 amino acids to about 70 amino acids, about 30 amino acids to about 60 amino acids, about 30 amino acids to about 50 amino acids, about 30 amino acids to about 45 amino acids, about 30 amino acids to about 40 amino acids , about 30 amino acids to about 35 amino acids, about 35 amino acids to about 1400 amino acids, about 35 amino acids to about 1350 amino acids, about 35 amino acids to about 1300 amino acids, about 35 amino acids to about 1250 amino acids, about 35 amino acids to about 1200 amino acids, about 35 amino acids to about 1150 amino acids, about 35 amino acids to about 1100 amino acids, about 35 amino acids to about 1050 amino acids, about 35 amino acids to about 1000 amino acids, about 35 amino acids to about 950 amino acids, about 35 amino acids to about 900 amino acids, about 35 amino acids to about 850 amino acids, about 35 amino acids to about 800 amino acids, from about 35 amino acids to about 750 amino acids, from about 35 amino acids to about 700 amino acids, from about 35 amino acids to about 650 amino acids, from about 35 amino acids to about 600 amino acids, from about 35 amino acids to about 550 amino acids, from about 35 amino acids to about 500 amino acids, from about 35 amino acids to about 450 amino acids, from about 35 amino acids to about 400 amino acids, from about 35 amino acids to about 350 amino acids, from about 35 amino acids to about 300 amino acids, from about 35 amino acids to about 250 amino acids, from about 35 amino acids to about 200 amino acids, from about 35 amino acids to about 150 amino acids, from about 35 amino acids to about 100 amino acids, from about 35 amino acids to about from about 90 amino acids, from about 35 amino acids to about 80 amino acids, from about 35 amino acids to about 70 amino acids, from about 35 amino acids to about 60 amino acids, from about 35 amino acids to about 50 amino acids, from about 35 amino acids to about 45 amino acids, from about 35 amino acids to about 40 amino acids, from about 40 amino acids to about 1400 amino acids, from about 40 amino acids to about 1350 amino acids, from about 40 amino acids to about 1300 amino acids, from about 40 amino acids to about 1250 amino acids, from about 40 amino acids to about 1200 amino acids, from about 40 amino acids to about 1150 amino acids, from about 40 amino acids to about 1100 amino acids, from about 40 amino acids to about 1050 amino acids,about 40 amino acids to about 1000 amino acids, about 40 amino acids to about 950 amino acids, about 40 amino acids to about 900 amino acids, about 40 amino acids to about 850 amino acids, about 40 amino acids to about 800 amino acids, about 40 amino acids to about 750 amino acids, about 40 amino acids to about 700 amino acids, about 40 amino acids to about 650 amino acids, about 40 amino acids to about 600 amino acids, about 40 amino acids to about 550 amino acids, about 40 amino acids to about 500 amino acids, about 40 amino acids to about 450 amino acids, about 40 amino acids to about 400 amino acids, about 40 amino acids to about 350 amino acids, about 40 amino acids to about 350 amino acids, about 40 amino acids to about 500 amino acids. about 40 amino acids to about 300 amino acids, about 40 amino acids to about 250 amino acids, about 40 amino acids to about 200 amino acids, about 40 amino acids to about 150 amino acids, about 40 amino acids to about 100 amino acids, about 40 amino acids to about 90 amino acids, about 40 amino acids to about 80 amino acids, about 40 amino acids to about 70 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 45 amino acids to about 1400 amino acids, about 45 amino acids to about 1350 amino acids, about 45 amino acids to about 1300 amino acids, about 45 amino acids to about 1250 amino acids, about about 45 amino acids to about 1200 amino acids, about 45 amino acids to about 1150 amino acids, about 45 amino acids to about 1100 amino acids, about 45 amino acids to about 1050 amino acids, about 45 amino acids to about 1000 amino acids, about 45 amino acids to about 950 amino acids, about 45 amino acids to about 900 amino acids, about 45 amino acids to about 850 amino acids, about 45 amino acids to about 800 amino acids, about 45 amino acids to about 750 amino acids, about 45 amino acids to about 700 amino acids, about 45 amino acids to about 650 amino acids, about 45 amino acids to about 600 amino acids, about 45 amino acids to about 550 amino acids, about 45 from about 45 amino acids to about 500 amino acids, from about 45 amino acids to about 450 amino acids, from about 45 amino acids to about 400 amino acids, from about 45 amino acids to about 350 amino acids, from about 45 amino acids to about 300 amino acids, from about 45 amino acids to about 250 amino acids, from about 45 amino acids to about 200 amino acids, from about 45 amino acids to about 150 amino acids, from about 45 amino acids to about 100 amino acids, from about 45 amino acids to about 90 amino acids, from about 45 amino acids to about 80 amino acids, from about 45 amino acids to about 70 amino acids, from about 45 amino acids to about 60 amino acids, from about 45 amino acids to about 50 amino acids, from about 50 amino acids to about 1400 amino acids,about 50 amino acids to about 1350 amino acids, about 50 amino acids to about 1300 amino acids, about 50 amino acids to about 1250 amino acids, about 50 amino acids to about 1200 amino acids, about 50 amino acids to about 1150 amino acids, about 50 amino acids to about 1100 amino acids, about 50 amino acids to about 1050 amino acids, about 50 amino acids to about 1000 amino acids, about 50 amino acids to about 950 amino acids, about 50 amino acids to about 900 amino acids, about 50 amino acids to about 850 amino acids, about 50 amino acids to about 800 amino acids, about 50 amino acids to about 750 amino acids, about 50 amino acids to about 700 amino acids, about 50 From about 50 amino acids to about 650 amino acids, from about 50 amino acids to about 600 amino acids, from about 50 amino acids to about 550 amino acids, from about 50 amino acids to about 500 amino acids, from about 50 amino acids to about 450 amino acids, from about 50 amino acids to about 400 amino acids, from about 50 amino acids to about 350 amino acids, from about 50 amino acids to about 300 amino acids, from about 50 amino acids to about 250 amino acids, from about 50 amino acids to about 200 amino acids, from about 50 amino acids to about 150 amino acids, from about 50 amino acids to about 100 amino acids, from about 50 amino acids to about 90 amino acids, from about 50 amino acids to about 80 amino acids, from about 50 amino acids to about 70 amino acids, from about 50 amino acids to about From about 50 amino acids to about 60 amino acids, from about 60 amino acids to about 1400 amino acids, from about 60 amino acids to about 1350 amino acids, from about 60 amino acids to about 1300 amino acids, from about 60 amino acids to about 1250 amino acids, from about 60 amino acids to about 1200 amino acids, from about 60 amino acids to about 1150 amino acids, from about 60 amino acids to about 1100 amino acids, from about 60 amino acids to about 1050 amino acids, from about 60 amino acids to about 1000 amino acids, from about 60 amino acids to about 950 amino acids, from about 60 amino acids to about 900 amino acids, from about 60 amino acids to about 850 amino acids, from about 60 amino acids to about 800 amino acids, from about 60 amino acids to about 750 amino acids, about 60 amino acids to about 700 amino acids, about 60 amino acids to about 650 amino acids, about 60 amino acids to about 600 amino acids, about 60 amino acids to about 550 amino acids, about 60 amino acids to about 500 amino acids, about 60 amino acids to about 450 amino acids, about 60 amino acids to about 400 amino acids, about 60 amino acids to about 350 amino acids, about 60 amino acids to about 300 amino acids, about 60 amino acids to about 250 amino acids, about 60 amino acids to about 200 amino acids, about 60 amino acids to about 150 amino acids, about 60 amino acids to about 100 amino acids, about 60 amino acids to about 90 amino acids,about 60 amino acids to about 80 amino acids, about 60 amino acids to about 70 amino acids, about 70 amino acids to about 1400 amino acids, about 70 amino acids to about 1350 amino acids, about 70 amino acids to about 1300 amino acids, about 70 amino acids to about 1250 amino acids, about 70 amino acids to about 1200 amino acids, about 70 amino acids to about 1150 amino acids, about 70 amino acids to about 1100 amino acids, about 70 amino acids to about 1050 amino acids, about 70 amino acids to about 1000 amino acids, about 70 amino acids to about 950 amino acids, about 70 amino acids to about 900 amino acids, about 70 amino acids to about 850 amino acids, about 70 amino acids to about 1 about 70 amino acids to about 800 amino acids, about 70 amino acids to about 750 amino acids, about 70 amino acids to about 700 amino acids, about 70 amino acids to about 650 amino acids, about 70 amino acids to about 600 amino acids, about 70 amino acids to about 550 amino acids, about 70 amino acids to about 500 amino acids, about 70 amino acids to about 450 amino acids, about 70 amino acids to about 400 amino acids, about 70 amino acids to about 350 amino acids, about 70 amino acids to about 300 amino acids, about 70 amino acids to about 250 amino acids, about 70 amino acids to about 200 amino acids, about 70 amino acids to about 150 amino acids, about 70 amino acids to about 100 amino acids, about 70 amino acids to about 90 amino acids, about 70 amino acids to about 80 amino acids, about 80 amino acids to about 1400 amino acids, about 80 amino acids to about 1350 amino acids, about 80 amino acids to about 1300 amino acids, about 80 amino acids to about 1250 amino acids, about 80 amino acids to about 1200 amino acids, about 80 amino acids to about 1150 amino acids, about 80 amino acids to about 1100 amino acids, about 80 amino acids to about 1050 amino acids, about 80 amino acids to about 1000 amino acids, about 80 amino acids to about 950 amino acids, about 80 amino acids to about 900 amino acids, about 80 amino acids to about 850 amino acids, about 80 amino acids to about 1 about 80 amino acids to about 800 amino acids, about 80 amino acids to about 750 amino acids, about 80 amino acids to about 700 amino acids, about 80 amino acids to about 650 amino acids, about 80 amino acids to about 600 amino acids, about 80 amino acids to about 550 amino acids, about 80 amino acids to about 500 amino acids, about 80 amino acids to about 450 amino acids, about 80 amino acids to about 400 amino acids, about 80 amino acids to about 350 amino acids, about 80 amino acids to about 300 amino acids, about 80 amino acids to about 250 amino acids, about 80 amino acids to about 200 amino acids, about 80 amino acids to about 150 amino acids, about 80 amino acids to about 100 amino acids,From about 80 amino acids to about 90 amino acids, from about 90 amino acids to about 1400 amino acids, from about 90 amino acids to about 1350 amino acids, from about 90 amino acids to about 1300 amino acids, from about 90 amino acids to about 1250 amino acids, from about 90 amino acids to about 1200 amino acids, from about 90 amino acids to about 1150 amino acids, from about 90 amino acids to about 1100 amino acids, from about 90 amino acids to about 1050 amino acids, from about 90 amino acids to about 1000 amino acids, from about 90 amino acids to about 950 amino acids, from about 90 amino acids to about 900 amino acids, from about 90 amino acids to about 850 amino acids, from about 90 amino acids to about 800 amino acids, from about 90

[0013] The present invention relates to a polypeptide comprising a polypeptide comprising a polypeptide of the present invention and a nucleotide sequence of the present invention. The polypeptide comprises a polypeptide of the present invention and a nucleotide sequence of the present invention. The polypeptide comprises a polypeptide of the present invention and a nucleotide sequence of the present invention. amino acids, about 100 amino acids to about 1350 amino acids, about 100 amino acids to about 1300 amino acids, about 100 amino acids to about 1250 amino acids, about 100 amino acids to about 1200 amino acids, about 100 amino acids to about 1150 amino acids, about 100 amino acids to about 1100 amino acids, about 100 amino acids to about 1050 amino acids, about 100 amino acids to about 1000 amino acids, about 100 amino acids to about 950 amino acids, about 100 amino acids to about 900 amino acids, about 100 amino acids to about 850 amino acids, about 100 amino acids to about 800 amino acids, about 100 amino acids to about 750 amino acids, about 100 from about 100 amino acids to about 700 amino acids, from about 100 amino acids to about 650 amino acids, from about 100 amino acids to about 600 amino acids, from about 100 amino acids to about 550 amino acids, from about 100 amino acids to about 500 amino acids, from about 100 amino acids to about 450 amino acids, from about 100 amino acids to about 400 amino acids, from about 100 amino acids to about 350 amino acids, from about 100 amino acids to about 300 amino acids, from about 100 amino acids to about 250 amino acids, from about 100 amino acids to about 200 amino acids, from about 100 amino acids to about 150 amino acids, from about 150 amino acids to about 1400 amino acids, from about 150 amino acids to about 1350 amino acids,about 150 amino acids to about 1300 amino acids, about 150 amino acids to about 1250 amino acids, about 150 amino acids to about 1200 amino acids, about 150 amino acids to about 1150 amino acids, about 150 amino acids to about 1100 amino acids, about 150 amino acids to about 1050 amino acids, about 150 amino acids to about 1000 amino acids, about 150 amino acids to about 950 amino acids, about 150 amino acids to about 900 amino acids, about 150 amino acids to about 850 amino acids, about 150 amino acids to about 800 amino acids, about 150 amino acids to about 750 amino acids, about 150 amino acids to about 700 amino acids, about 150 amino acids to about from about 150 amino acids to about 650 amino acids, from about 150 amino acids to about 600 amino acids, from about 150 amino acids to about 550 amino acids, from about 150 amino acids to about 500 amino acids, from about 150 amino acids to about 450 amino acids, from about 150 amino acids to about 400 amino acids, from about 150 amino acids to about 350 amino acids, from about 150 amino acids to about 300 amino acids, from about 150 amino acids to about 250 amino acids, from about 150 amino acids to about 200 amino acids, from about 200 amino acids to about 1400 amino acids, from about 200 amino acids to about 1350 amino acids, from about 200 amino acids to about 1300 amino acids, from about 200 amino acids to about 1250 amino acids, about 200 amino acids to about 1200 amino acids, about 200 amino acids to about 1150 amino acids, about 200 amino acids to about 1100 amino acids, about 200 amino acids to about 1050 amino acids, about 200 amino acids to about 1000 amino acids, about 200 amino acids to about 950 amino acids, about 200 amino acids to about 900 amino acids, about 200 amino acids to about 850 amino acids, about 200 amino acids to about 800 amino acids, about 200 amino acids to about 750 amino acids, about 200 amino acids to about 700 amino acids, about 200 amino acids to about 650 amino acids, about 200 amino acids to about 600 amino acids, about 200 amino acids to about from about 200 amino acids to about 500 amino acids, from about 200 amino acids to about 450 amino acids, from about 200 amino acids to about 400 amino acids, from about 200 amino acids to about 350 amino acids, from about 200 amino acids to about 300 amino acids, from about 200 amino acids to about 250 amino acids, from about 250 amino acids to about 1400 amino acids, from about 250 amino acids to about 1350 amino acids, from about 250 amino acids to about 1300 amino acids, from about 250 amino acids to about 1250 amino acids, from about 250 amino acids to about 1200 amino acids, from about 250 amino acids to about 1150 amino acids, from about 250 amino acids to about 1100 amino acids,from about 250 amino acids to about 1050 amino acids, from about 250 amino acids to about 1000 amino acids, from about 250 amino acids to about 950 amino acids, from about 250 amino acids to about 900 amino acids, from about 250 amino acids to about 850 amino acids, from about 250 amino acids to about 800 amino acids, from about 250 amino acids to about 750 amino acids, from about 250 amino acids to about 700 amino acids, from about 250 amino acids to about 650 amino acids, from about 250 amino acids to about 600 amino acids, from about 250 amino acids to about 550 amino acids, from about 250 amino acids to about 500 amino acids, from about 250 amino acids to about 450 amino acids, from about 250 amino acids to about 40 0 amino acids, about 250 amino acids to about 350 amino acids, about 250 amino acids to about 300 amino acids, about 300 amino acids to about 1400 amino acids, about 300 amino acids to about 1350 amino acids, about 300 amino acids to about 1300 amino acids, about 300 amino acids to about 1250 amino acids, about 300 amino acids to about 1200 amino acids, about 300 amino acids to about 1150 amino acids, about 300 amino acids to about 1100 amino acids, about 300 amino acids to about 1050 amino acids, about 300 amino acids to about 1000 amino acids, about 300 amino acids to about 950 amino acids, about 300 amino acids to about 900 amino acids. , about 300 amino acids to about 850 amino acids, about 300 amino acids to about 800 amino acids, about 300 amino acids to about 750 amino acids, about 300 amino acids to about 700 amino acids, about 300 amino acids to about 650 amino acids, about 300 amino acids to about 600 amino acids, about 300 amino acids to about 550 amino acids, about 300 amino acids to about 500 amino acids, about 300 amino acids to about 450 amino acids, about 300 amino acids to about 400 amino acids, about 300 amino acids to about 350 amino acids, about 350 amino acids to about 1400 amino acids, about 350 amino acids to about 1350 amino acids, about 350 amino acids to about 1 about 350 amino acids to about 1250 amino acids, about 350 amino acids to about 1200 amino acids, about 350 amino acids to about 1150 amino acids, about 350 amino acids to about 1100 amino acids, about 350 amino acids to about 1050 amino acids, about 350 amino acids to about 1000 amino acids, about 350 amino acids to about 950 amino acids, about 350 amino acids to about 900 amino acids, about 350 amino acids to about 850 amino acids, about 350 amino acids to about 800 amino acids, about 350 amino acids to about 750 amino acids, about 350 amino acids to about 700 amino acids, about 350 amino acids to about 650 amino acids,about 350 amino acids to about 600 amino acids, about 350 amino acids to about 550 amino acids, about 350 amino acids to about 500 amino acids, about 350 amino acids to about 450 amino acids, about 350 amino acids to about 400 amino acids, about 400 amino acids to about 1400 amino acids, about 400 amino acids to about 1350 amino acids, about 400 amino acids to about 1300 amino acids, about 400 amino acids to about 1250 amino acids, about 400 amino acids to about 1200 amino acids, about 400 amino acids to about 1150 amino acids, about 400 amino acids to about 1100 amino acids, about 400 amino acids to about 1050 amino acids, about 400 amino acids to about 1500 amino acids, about 400 amino acids to about 1050 amino acids, about 400 amino acids to about 1500 amino acids, about 400 amino acids to about 1050 amino acids, about 400 amino acids to about 1500 amino acids, about 400 amino acids to about 1050 amino acids, about 400 amino acids to about 1500 amino acids. The present invention relates to a polypeptide having at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. The polypeptide has at least one amino acid sequence and at least one amino acid sequence. about 450 amino acids to about 1300 amino acids, about 450 amino acids to about 1250 amino acids, about 450 amino acids to about 1200 amino acids, about 450 amino acids to about 1150 amino acids, about 450 amino acids to about 1100 amino acids, about 450 amino acids to about 1050 amino acids, about 450 amino acids to about 1000 amino acids, about 450 amino acids to about 950 amino acids, about 450 amino acids to about 900 amino acids, about 450 amino acids to about 850 amino acids, about 450 amino acids to about 800 amino acids, about 450 amino acids to about 750 amino acids, about 450 amino acids to about 700 amino acids, about 450 amino acids to about 6 from about 500 amino acids to about 1000 amino acids, from about 500 amino acids to about 1500 amino acids, from about 500 amino acids to about 1600 amino acids, from about 500 amino acids to about 1700 amino acids, from about 500 amino acids to about 1800 amino acids, from about 500 amino acids to about 1900 amino acids, from about 500 amino acids to about 1900 amino acids, from about 500 amino acids to about 1800 amino acids, from about 500 amino acids to about 1900 amino acids, from about 500 amino acids to about 1800 amino acids, from about 500 amino acids to about 1800 amino acids, from about 500 amino acids to about 1900 amino acids,from about 500 amino acids to about 900 amino acids, from about 500 amino acids to about 850 amino acids, from about 500 amino acids to about 800 amino acids, from about 500 amino acids to about 750 amino acids, from about 500 amino acids to about 700 amino acids, from about 500 amino acids to about 650 amino acids, from about 500 amino acids to about 600 amino acids, from about 500 amino acids to about 550 amino acids, from about 550 amino acids to about 1400 amino acids, from about 550 amino acids to about 1350 amino acids, from about 550 amino acids to about 1300 amino acids, from about 550 amino acids to about 1250 amino acids, from about 550 amino acids to about 1200 amino acids, from about 550 amino acids to about about 550 amino acids to about 1150 amino acids, about 550 amino acids to about 1100 amino acids, about 550 amino acids to about 1050 amino acids, about 550 amino acids to about 1000 amino acids, about 550 amino acids to about 950 amino acids, about 550 amino acids to about 900 amino acids, about 550 amino acids to about 850 amino acids, about 550 amino acids to about 800 amino acids, about 550 amino acids to about 750 amino acids, about 550 amino acids to about 700 amino acids, about 550 amino acids to about 650 amino acids, about 550 amino acids to about 600 amino acids, about 600 amino acids to about 1400 amino acids, about 600 amino acids to about 1350 amino acids, about 600 amino acids to about 1500 amino acids, about 600 amino acids to about 1600 amino acids, about 600 amino acids to about 1700 amino acids, about 600 amino acids to about 1800 amino acids, about 600 amino acids to about 1900 amino acids, about 600 amino acids to about 1950 amino acids, about 600 amino acids to about 1960 amino acids, about 600 amino acids to about 1970 amino acids, about 600 amino acids to about 1980 amino acids, about 600 amino acids to about 19 ... 00 amino acids to about 1300 amino acids, about 600 amino acids to about 1250 amino acids, about 600 amino acids to about 1200 amino acids, about 600 amino acids to about 1150 amino acids, about 600 amino acids to about 1100 amino acids, about 600 amino acids to about 1050 amino acids, about 600 amino acids to about 1000 amino acids, about 600 amino acids to about 950 amino acids, about 600 amino acids to about 900 amino acids, about 600 amino acids to about 850 amino acids, about 600 amino acids to about 800 amino acids, about 600 amino acids to about 750 amino acids, about 600 amino acids to about 700 amino acids, about 600 amino acids to about 6 about 650 amino acids to about 1500 amino acids, about 650 amino acids to about 1400 amino acids, about 650 amino acids to about 1350 amino acids, about 650 amino acids to about 1300 amino acids, about 650 amino acids to about 1250 amino acids, about 650 amino acids to about 1200 amino acids, about 650 amino acids to about 1150 amino acids, about 650 amino acids to about 1100 amino acids, about 650 amino acids to about 1050 amino acids, about 650 amino acids to about 1000 amino acids, about 650 amino acids to about 950 amino acids, about 650 amino acids to about 900 amino acids, about 650 amino acids to about 850 amino acids, about 650 amino acids to about 800 amino acids,about 650 amino acids to about 750 amino acids, about 650 amino acids to about 700 amino acids, about 700 amino acids to about 1400 amino acids, about 700 amino acids to about 1350 amino acids, about 700 amino acids to about 1300 amino acids, about 700 amino acids to about 1250 amino acids, about 700 amino acids to about 1200 amino acids, about 700 amino acids to about 1150 amino acids, about 700 amino acids to about 1100 amino acids, about 700 amino acids to about 1050 amino acids, about 700 amino acids to about 1000 amino acids, about 700 amino acids to about 950 amino acids, about 700 amino acids to about 900 amino acids, about 700 amino acids to about 850 amino acids, about 700 amino acids to about 800 amino acids, about 700 amino acids to about 750 amino acids, about 750 amino acids to about 1400 amino acids, about 750 amino acids to about 1350 amino acids, about 750 amino acids to about 1300 amino acids, about 750 amino acids to about 1250 amino acids, about 750 amino acids to about 1200 amino acids, about 750 amino acids to about 1150 amino acids, about 750 amino acids to about 1100 amino acids, about 750 amino acids to about 1050 amino acids, about 750 amino acids to about 1000 amino acids, about 750 amino acids to about 950 amino acids, about 750 amino acids to about 1 about 900 amino acids, about 750 amino acids to about 850 amino acids, about 750 amino acids to about 800 amino acids, about 800 amino acids to about 1400 amino acids, about 800 amino acids to about 1350 amino acids, about 800 amino acids to about 1300 amino acids, about 800 amino acids to about 1250 amino acids, about 800 amino acids to about 1200 amino acids, about 800 amino acids to about 1150 amino acids, about 800 amino acids to about 1100 amino acids, about 800 amino acids to about 1050 amino acids, about 800 amino acids to about 1000 amino acids, about 800 amino acids to about 950 amino acids, about 800 amino acids to about 900 amino acids, about 800 amino acids to about 850 amino acids, about 850 amino acids to about 1400 amino acids, about 850 amino acids to about 1350 amino acids, about 850 amino acids to about 1300 amino acids, about 850 amino acids to about 1250 amino acids, about 850 amino acids to about 1200 amino acids, about 850 amino acids to about 1150 amino acids, about 850 amino acids to about 1100 amino acids, about 850 amino acids to about 1050 amino acids, about 850 amino acids to about 1000 amino acids, about 850 amino acids to about 950 amino acids, about 850 amino acids to about 900 amino acids, about 900 amino acids to about 1400 amino acids,about 900 amino acids to about 1350 amino acids, about 900 amino acids to about 1300 amino acids, about 900 amino acids to about 1250 amino acids, about 900 amino acids to about 1200 amino acids, about 900 amino acids to about 1150 amino acids, about 900 amino acids to about 1100 amino acids, about 900 amino acids to about 1050 amino acids, about 900 amino acids to about 1000 amino acids, about 900 amino acids to about 950 amino acids, about 950 amino acids to about 1400 amino acids, about 950 amino acids to about 1350 amino acids, about 950 amino acids to about 1300 amino acids, about 950 amino acids to about 1250 amino acids, about 950 amino acids to about 1200 amino acids, about 950 amino acids to about 1150 amino acids, about 950 amino acids to about 1100 amino acids, about 950 amino acids to about 1050 amino acids, about 950 amino acids to about 1000 amino acids, about 1000 amino acids to about 1400 amino acids, about 1000 amino acids to about 1350 amino acids, about 1000 amino acids to about 1300 amino acids, about 1000 amino acids to about 1250 amino acids, about 1000 amino acids to about 1200 amino acids, about 1000 amino acids to about 1150 amino acids, about 1000 amino acids to about 1100 amino acids, about 1000 amino acids to about 1050 amino acids. , about 1050 amino acids to about 1400 amino acids, about 1050 amino acids to about 1350 amino acids, about 1050 amino acids to about 1300 amino acids, about 1050 amino acids to about 1250 amino acids, about 1050 amino acids to about 1200 amino acids, about 1050 amino acids to about 1150 amino acids, about 1050 amino acids to about 1100 amino acids, about 1100 amino acids to about 1400 amino acids, about 1100 amino acids to about 1350 amino acids, about 1100 amino acids to about 1300 amino acids, about 1100 amino acids to about 1250 amino acids, about 1100 amino acids to about 1200 amino acids, about 1100 amino acids about 1150 amino acids to about 1400 amino acids, about 1150 amino acids to about 1350 amino acids, about 1150 amino acids to about 1300 amino acids, about 1150 amino acids to about 1250 amino acids, about 1150 amino acids to about 1200 amino acids, about 1200 amino acids to about 1400 amino acids, about 1200 amino acids to about 1350 amino acids, about 1200 amino acids to about 1300 amino acids, about 1200 amino acids to about 1250 amino acids, about 1250 amino acids to about 1400 amino acids, about 1250 amino acids to about 1350 amino acids, about 1250 amino acids to about 1300 amino acids,From about 1300 amino acids to about 1400 amino acids, from about 1300 amino acids to about 1350 amino acids, or from about 1350 amino acids to about 1400 amino acids.

[0280] For example, the β-TCP binding polypeptide may include the amino acid sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRPW GILAESTHHKPWTATGGSGEGGTGASTGGSAGTGGSGGTTSGEAGG SSGAG (SEQ ID NO: 44). In some cases, the β-TCP binding polypeptide includes an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) sequence identity to SEQ ID NO: 44.

[0281] For example, the β-TCP binding polypeptide may include: LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHHR PWGLLADTTHHRPWTTGGSGEGGTGASTGGSAGTGGSGGTTSGEAG GSSGAG (SEQ ID NO: 45). In some cases, the β-TCP binding polypeptide includes an amino acid sequence having at least 70% (e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) sequence identity to SEQ ID NO: 45.

[0282] The K of any β-TCP binding polypeptide described herein for binding to β-TCP (e.g., any type of β-TCP described herein) can be determined using a variety of different methods known in the art. D values ​​(e.g., electrophoretic mobility shift assays, filter binding assays, surface plasmon resonance, and biomolecule binding kinetics assays, etc.).

[0283] Nucleic acid / vector

[0284] Also provided herein are nucleic acids encoding any of the chimeric polypeptides or β-TCP binding polypeptides described herein.

[0285] Also provided herein are vectors comprising any nucleic acid provided herein. A "vector" according to the present disclosure is a polynucleotide capable of inducing expression of a recombinant protein (e.g., any chimeric polypeptide or β-TCP binding polypeptide described) in a host cell. The vectors provided herein can be in, for example, circular or linearized form. Non-limiting examples of vectors include plasmids, SV40 vectors, adenoviral vectors, and adeno-associated virus (AAV) vectors. Non-limiting examples of vectors include lentiviral vectors or retroviral vectors, such as γ-retroviral vectors. See, for example, Carlens et al., Exp. Hematol. 28 (10): 1137-1146, 2000; Park et al., Trends Biotechnol. 29 (11): 550-557, 2011; and Alonso-Camino et al., Mol. Ther. Nucleic Acids 2: e93, 2013. Non-limiting examples of retroviral vectors include those derived from Moloney murine leukemia virus, myeloproliferative sarcoma virus, mouse embryonic stem cell virus, mouse stem cell virus, spleen focus forming virus or adeno-associated virus. Non-limiting examples of retroviral vectors are described in, for example, U.S. Pat. Nos. 5,219,740 and 6,207,453; Miller et al., BioTechniques 7:980-990, 1989; Miller et al., Human Gene Therapy 1:5-14, 1990; Scarpa et al., Virology 180:849-852, 1991; Burns et al., Proc. Natl. Acad. Sci. USA 90:8033-8037, 1993; and Boris-Lawrie et al., Cur. Opin. Genet. Develop. 3:102-109, 1993. Exemplary lentiviral vectors are described, e.g., in Wang et al., J. Immunother. 35(9):689-701, 2003; Cooper et al., Blood 101:1637-1644, 2003; Verhoeyen et al., Methods Mol. Biol. 506:97-114, 2009; and Cavalieri et al., Blood 102(2):497-505, 2003.

[0286] Exemplary vectors into which any of the nucleic acids provided herein can be inserted are described, e.g., in Ausubel et al., ed., "Current Protocols in Molecular Biology" Current Protocols, 1993; and Sambrook et al., ed., "Molecular Cloning: A Laboratory Manual", 2nd edition, Cold Spring Harbor Press, 1989.

[0287] In some embodiments, the vector further comprises a promoter and / or enhancer operably linked to any of the nucleic acids described herein. Non-limiting examples of promoters include promoters from human cytomegalovirus (CMV), mouse phosphoglycerate kinase 1, polyoma adenovirus, thyroid stimulating hormone alpha, vimentin, simian virus 40 (SV40), tumor necrosis factor, beta-globulin, alpha-fetoprotein, gamma-globulin, beta-interferon, gamma-glutamyl transferase, human ubiquitin C (UBC), mouse mammary tumor virus (MMTV), Rous sarcoma virus, glyceraldehyde-3-phosphate dehydrogenase, beta-actin, metallothionein II (MT II), amylase, human EF1α, cathepsin, MI muscarinic receptor, retroviral LTR (e.g., human T-cell leukemia virus HTLV), AAV ITR, interleukin-2, collagenase, platelet-derived growth factor, adenovirus E2, mesenchymal lysin, murine MX, rat insulin, glucose-regulated protein 78, human immunodeficiency virus, glucose-regulated protein 94, alpha-2-macroglobulin, class I MHC, HSP70, tetronic globulin, immunoglobulin light chain, T cell receptor, HLA DQα, HLA DQβ, interleukin-2 receptor, MHC class II, prealbumin (transthyretin), elastase I, albumin, c-fos, neural cell adhesion molecule (NCAM), H2B histone, rat growth hormone, human serum amyloid protein (SAA), muscle creatinine kinase, troponin I (TN I), and gibbon ape leukemia virus (GALV) promoter. In some embodiments, the promoter can be an inducible promoter or a constitutive promoter. Additional examples of promoters are known in the art.

[0288] In some examples, the vectors provided herein further include a poly(A) sequence operably linked and positioned 3' to the sequence encoding the chimeric polypeptide or the β-TCP binding polypeptide. Non-limiting examples of poly(A) sequences include those derived from bovine growth hormone (Woychik et al., Proc. Natl. Acad. Sci. USA 81(13):3944-3948, 1984, and U.S. Pat. No. 5,122,458), mouse-β-globulin, mouse-α-globulin (Orkin et al., EMBO J. 4(2):453-456, 1985), human collagen, polyomavirus (Batt et al., Mol. Cell Biol. 15(9):4783-4790, 1995), herpes simplex virus thymidine kinase gene (HSV TK), IgG heavy chain gene polyadenylation signal (U.S. Patent Application Publication No. 2006 / 0040354), human growth hormone (hGH) (Szymanski et al., Mol. Therapy 15(7):1340-1347, 2007), SV40 poly(A) site, for example, SV40 late and early poly(A) sites (Schek et al., Mol. Cell Biol. 12(12):5386-5393, 1992). In some embodiments, the poly(A) sequence includes a highly conserved upstream element (AATAAA). The AATAAA sequence can be replaced, for example, by other hexanucleotide sequences having homology to AATAAA, which are capable of emitting polyadenylation signals, including, for example, ATTAAA, AGTAAA, CATAAA, TATAAA, GATAAA, ACTAAA, AATATA, AAGAAA, AATGAAT, AAAAAA, AATGAA, AATCAA, AACAAA, AATCAA, AATTAAC, AATTAGA, AATTAAA, and AATAAG. See, for example, WO 06012414A2). The poly(A) sequence can be, for example, a synthetic polyadenylation site. See, e.g., Levitt et al., Genes Dev. 3(7): 1019-1025, 1989). In some examples, the poly(A) sequence can be the polyadenylation signal of soluble neuropilin-1: AAATAAAATACGAAATG. Additional examples of poly(A) sequences are known in the art. Other examples and aspects of vectors are also known in the art.

[0289] Methods for preparing chimeric polypeptides

[0290] Also provided herein is a method for preparing a chimeric polypeptide (e.g., any chimeric polypeptide described herein) or a β-TCP binding polypeptide (e.g., any β-TCP binding polypeptide described herein), comprising: introducing a nucleic acid sequence encoding the chimeric polypeptide or β-TCP binding polypeptide into a cell to produce a recombinant cell; and culturing the recombinant cell under conditions sufficient to express the chimeric polypeptide or β-TCP binding polypeptide. In some embodiments, the introducing step comprises introducing an expression vector into the cell to produce a recombinant cell, the expression vector comprising a nucleic acid sequence encoding the chimeric polypeptide or β-TCP binding polypeptide. In some embodiments, the expression vector comprises a chaperone (e.g., GroES, GroEL) and glutathione to assist in folding in vitro.

[0291] The chimeric polypeptides or β-TCP binding polypeptides described herein can be produced by any cell, such as a eukaryotic cell or a prokaryotic cell. As used herein, the term "eukaryotic cell" refers to a cell having a different membrane-bound nucleus. Such cells may include, for example, mammalian (e.g., rodent, non-human primate or human), insect, fungal or plant cells. In some embodiments, the eukaryotic cell is a yeast cell, such as Saccharomyces cerevisiae. In some embodiments, the eukaryotic cell is a higher eukaryotic organism, such as a mammalian, avian, plant or insect cell. In some embodiments, the eukaryotic cell is a mammalian cell (e.g., Chinese hamster ovary (CHO) cell). As used herein, the term "prokaryotic cell" refers to a cell that does not have a different membrane-bound nucleus. In some embodiments, the prokaryotic cell is a bacterial cell. In some embodiments, the bacterial cell is a chemically competent Escherichia coli K12 cell (e.g., T7; New England BioLabs) or BL21(DE3) pLysS chemically competent E. coli cells.

[0292] Methods for culturing cells are well known in the art. Cells can be maintained in vitro under conditions that are conducive to proliferation, differentiation and growth. In short, cells can be cultured by contacting cells (e.g., any cells) with a cell culture medium that includes essential growth factors and supplements to support cell survival and growth.

[0293] Methods for introducing nucleic acids and expression vectors into cells (e.g., eukaryotic cells) are known in the art. Non-limiting examples of methods that can be used to introduce nucleic acids into cells include lipofection, transfection, electroporation, microinjection, calcium phosphate transfection, dendrimer-based transfection, cationic polymer transfection, cell squeezing, sonoporation, optical transfection, impalection, hydrodynamic delivery, magnetofection, viral transduction (e.g., adenoviral and lentiviral transduction), and nanoparticle transfection.

[0294] Provided herein are methods that also include isolating a chimeric polypeptide or β-TCP binding polypeptide from a cell (e.g., a eukaryotic cell) using techniques well known in the art (e.g., ammonium sulfate precipitation, polyethylene glycol precipitation, cobalt columns, heparin columns, ion exchange chromatography (anionic or cationic), hydrophobic interaction-based chromatography, metal-affinity chromatography, ligand-affinity chromatography, and size exclusion chromatography).

[0295] In some embodiments, a hydrophobicity / hydrophilicity plot is constructed to identify variant β-TCP binding sequences that may be included in any of the chimeric polypeptides described herein.

[0296] β-TCP

[0297] Sintering of tricalcium phosphate, Ca3(PO4)2, results in its structural conversion to β-TCP. β-TCP is an osteoconductive material that supports bone mineralization by readily dissolving at low pH and serves as a rigid substrate for cell attachment (see, e.g., Muschler et al., J. Bone Joint Surgery 86:1541-1558, 2004; Fleming Jr. et al., "Intraoperative Harvest and Concentration of Human Bone Marrow Osteoprogenitors for Enhancement of Spinal Fusion," in Orthopedic Tissue Engineering: Basic Science and Practice, 2004).

[0298] β-TCP as described herein can be in a variety of different forms. The example of such form includes a granular form, a porous form, a powder, a putty (e.g., a moldable putty), a paste, a stent and / or a coating on a solid surface (e.g., a coating on a medical device). In addition, β-TCP can be used for various shapes (e.g., a cross, a trapezoid, a sphere, an ellipse, a square, a triangular pyramid, a rod, a cone, an annular ring or a wedge or any combination thereof) and a size (e.g., a maximum average diameter of about 1 mm, about 2 mm, about 3 mm, about 4 mm, about 5 mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, about 1 cm, about 2 cm, about 3 cm, about 4 cm, about 5 cm, about 6 cm, about 7 cm, about 8 cm, about 9 cm or 10 cm). In some embodiments, β-TCP can be porous (e.g., about 20% to about 80% porous, about 20% to about 75% porous, about 20% to about 70% porous, about 20% to about 65% porous, about 20% to about 60% porous, about 20% to about 55% porous, about 20% to about 50% porous, about 20% to about 45% porous, about 20% to about 40% porous, about 20% to about 35% porous, about 20% to about 30% porous, about 20% to about 25% porous, about 25% to about 80% porous, about 25% to about 100% porous, about 25% to about 30% porous, about 20% to about 25% porous, about 25% to about 80% porous, about 25% to about 100% porous, about 25% to about 150% porous, about 25% to about 10 ... % to about 75% porous, about 25% to about 70% porous, about 25% to about 65% porous, about 25% to about 60% porous, about 25% to about 55% porous, about 25% to about 50% porous, about 25% to about 45% porous, about 25% to about 40% porous, about 25% to about 35% porous, about 25% to about 30% porous, about 30% to about 80% porous, about 30% to about 75% porous, about 30% to about 70% porous, about 30% to about 65% porous, about 30% to about 60% porous, about 30% to about about 30% to about 50% porous, about 30% to about 45% porous, about 30% to about 40% porous, about 30% to about 35% porous, about 35% to about 80% porous, about 35% to about 75% porous, about 35% to about 70% porous, about 35% to about 65% porous, about 35% to about 60% porous, about 35% to about 55% porous, about 35% to about 50% porous, about 35% to about 45% porous, about 35% to about 40% porous, about 40% to about 80% porous, about 40% to about 75% porous porous, about 40% to about 70% porous, about 40% to about 65% porous, about 40% to about 60% porous, about 40% to about 55% porous, about 40% to about 50% porous, about 40% to about 45% porous, about 45% to about 80% porous, about 45% to about 75% porous, about 45% to about 70% porous, about 45% to about 65% porous, about 45% to about 60% porous, about 45% to about 55% porous, about 45% to about 50% porous, about 50% to about 80% porous, about 50% to about 75% porous,about 50% to about 70% porous, about 50% to about 65% porous, about 50% to about 60% porous, about 50% to about 55% porous, about 55% to about 80% porous, about 55% to about 75% porous, about 55% to about 70% porous, about 55% to about 65% porous, about 55% to about 60% porous, about 60% to about 80% porous, about 60% to about 75% porous, about 60% to about 70% porous, about 60% to about 65% porous, about 65% to about 80% porous, about 65% to about 75% porous, about 65% to about 70% porous, about 70% to about 80% porous, about 70% to about 75% porous, or about 75% to about 80% porous). For example, the average pore size of β-TCP can be from about 1 nm to about 50 mm, from about 1 nm to about 45 mm, from about 1 nm to about 40 mm, from about 1 nm to about 35 mm, from about 1 nm to about 30 mm, from about 1 nm to about 25 mm, from about 1 nm to about 20 mm, from about 1 nm to about 15 mm, from about 1 nm to about 10 mm, from about 1 nm to about 5 mm, from about 1 nm to about 1 mm, from about 1 nm to about 900 μm, from about 1 nm to about 800 μm, from about 1 nm to about 700 μm, from about 1 nm to about 600 μm, from about 1 nm to about 50 0 μm, about 1 nm to about 400 μm, about 1 nm to about 300 μm, about 1 nm to about 200 μm, about 1 nm to about 100 μm, about 1 nm to about 50 μm, about 1 nm to about 10 μm, about 1 nm to about 5 μm, about 1 nm to about 1 μm, about 1 nm to about 900 nm, about 1 nm to about 800 nm, about 1 nm to about 700 nm, about 1 nm to about 600 nm, about 1 nm to about 500 nm, about 1 nm to about 400 nm, about 1 nm to about 300 nm, about 1 nm to about 200 nm, about 1 nm to about 100 nm, about 1 nm to about 50 nm, about 1 nm to about 25 nm, about 1 nm to about 10 nm, about 1 nm to about 5 nm, about 5 nm to about 50 mm, about 5 nm to about 45 mm, about 5 nm to about 40 mm, about 5 nm to about 35 mm, about 5 nm to about 30 mm, about 5 nm to about 25 mm, about 5 nm to about 20 mm, about 5 nm to about 15 mm, about 5 nm to about 10 mm, about 5 nm to about 5 mm, about 5 nm to about 1 mm, about 5 nm to about 900 μm, about 5 nm to about 800 μm, about 5 nm to about 700 μm, about 5 nm to about 600 μm, about 5 nm to about 500 μm, about 5 nm to about 400 μm, about 5 nm to about 300 μm, about 5 nm to about 200 μm, about 5 nm to about 100 μm, about 5 nm to about 50 μm, about 5 nm to about 10 μm, about 5 nm to about 5 μm, about 5 nm to about 1 μm, about 5 nm to about 900 nm, about 5 nm to about 800 nm, about 5 nm to about 700 nm, about 5 nm to about 600 nm, about 5 nm to about 500 nm,from about 5 nm to about 400 nm, from about 5 nm to about 300 nm, from about 5 nm to about 200 nm, from about 5 nm to about 100 nm, from about 5 nm to about 50 nm, from about 5 nm to about 25 nm, from about 5 nm to about 10 nm, from about 10 nm to about 50 mm, from about 10 nm to about 45 mm, from about 10 nm to about 40 mm, from about 10 nm to about 35 mm, from about 10 nm to about 30 mm, from about 10 nm to about 25 mm, from about 10 nm to about 20 mm, from about 10 nm to about 15 mm, from about 10 nm to about 10 mm, from about 10 nm to about 5 mm, from about 10 nm to about 1 mm, from about 10 nm to about 900 μm, from about 10 nm to about 800 μm, from about 10 nm to about 700 μm , about 10 nm to about 600 μm, about 10 nm to about 500 μm, about 10 nm to about 400 μm, about 10 nm to about 300 μm, about 10 nm to about 200 μm, about 10 nm to about 100 μm, about 10 nm to about 50 μm, about 10 nm to about 10 μm, about 10 nm to about 5 μm, about 10 nm to about 1 μm, about 10 nm to about 900 nm, about 10 nm to about 800 nm, about 10 nm to about 700 nm, about 10 nm to about 600 nm, about 10 nm to about 500 nm, about 10 nm to about 400 nm, about 10 nm to about 300 nm, about 10 nm to about 200 nm, about 10 nm to about 100 nm, about 10 nm to about 50nm, about 10nm to about 25nm, about 25nm to about 50mm, about 25nm to about 45mm, about 25nm to about 40mm, about 25nm to about 35mm, about 25nm to about 30mm, about 25nm to about 25mm, about 25nm to about 20mm, about 25nm to about 15mm, about 25nm to about 10mm, about 25nm to about 5mm, about 25nm to about 1mm, about 25nm to about 900μm, about 25nm to about 800μm, about 25nm to about 700μm, about 25nm to about 600μm, about 25nm to about 500μm, about 25nm to about 400μm, about 25nm to about 300μm, about 25nm to about 200μm m, about 25 nm to about 100 μm, about 25 nm to about 50 μm, about 25 nm to about 10 μm, about 25 nm to about 5 μm, about 25 nm to about 1 μm, about 25 nm to about 900 nm, about 25 nm to about 800 nm, about 25 nm to about 700 nm, about 25 nm to about 600 nm, about 25 nm to about 500 nm, about 25 nm to about 400 nm, about 25 nm to about 300 nm, about 25 nm to about 200 nm, about 25 nm to about 100 nm, about 25 nm to about 50 nm, about 50 nm to about 50 mm, about 50 nm to about 45 mm, about 50 nm to about 40 mm, about 50 nm to about 35 mm, about 50 nm to about 30 mm,about 50nm to about 25mm, about 50nm to about 20mm, about 50nm to about 15mm, about 50nm to about 10mm, about 50nm to about 5mm, about 50nm to about 1mm, about 50nm to about 900μm, about 50nm to about 800μm, about 50nm to about 700μm, about 50nm to about 600μm, about 50nm to about 500μm, about 50nm to about 400μm, about 50nm to about 300μm, about 50nm to about 200μm, about 50nm to about 100μm, about 50nm to about 50μm, about 50nm to about 10μm, about 50nm to about 5μm, about 50nm to about 1μm, about 50nm to about 900nm, about 50nm to about 800nm, about 50nm to about 700nm, about 50nm to about 600nm, about 50nm to about 500nm, about 50nm to about 400nm, about 50nm to about 300nm, about 50nm to about 200nm, about 50nm to about 100nm, about 100nm to about 50mm, about 100nm to about 45mm, about 100nm to about 40mm, about 100nm to about 35mm, about 100nm to about 30mm, about 100nm to about 25mm, about 100nm to about 20mm, about 100nm to about 15mm, about 100nm to about 10mm, about 100nm to about 5mm, about 100nm to about 1mm, about 100 nm to about 900 μm, about 100 nm to about 800 μm, about 100 nm to about 700 μm, about 100 nm to about 600 μm, about 100 nm to about 500 μm, about 100 nm to about 400 μm, about 100 nm to about 300 μm, about 100 nm to about 200 μm, about 100 nm to about 100 μm, about 100 nm to about 50 μm, about 100 nm to about 10 μm, about 100 nm to about 5 μm, about 100 nm to about 1 μm, about 100 nm to about 900 nm, about 100 nm to about 800 nm, about 100 nm to about 700 nm, about 100 nm to about 600 nm, about 100 nm to about 500 nm, about 100 nm to about m to about 400nm, about 100nm to about 300nm, about 100nm to about 200nm, about 200nm to about 50mm, about 200nm to about 45mm, about 200nm to about 40mm, about 200nm to about 35mm, about 200nm to about 30mm, about 200nm to about 25mm, about 200nm to about 20mm, about 200nm to about 15mm, about 200nm to about 10mm, about 200nm to about 5mm, about 200nm to about 1mm, about 200nm to about 900μm, about 200nm to about 800μm, about 200nm to about 700μm, about 200nm to about 600μm, about 200nm to about 500μm,about 200nm to about 400μm, about 200nm to about 300μm, about 200nm to about 200μm, about 200nm to about 100μm, about 200nm to about 50μm, about 200nm to about 10μm, about 200nm to about 5μm, about 200nm to about 1μm, about 200nm to about 900nm, about 200nm to about 800nm, about 200nm to about 700nm, about 200nm to about 600nm, about 200nm to about 500nm, about 200nm to about 400nm, about 200nm to about 300nm, about 300nm to about 50mm, about 300nm to about 45mm, about 300nm to about 40mm, about 300nm to about m to about 35mm, about 300nm to about 30mm, about 300nm to about 25mm, about 300nm to about 20mm, about 300nm to about 15mm, about 300nm to about 10mm, about 300nm to about 5mm, about 300nm to about 1mm, about 300nm to about 900μm, about 300nm to about 800μm, about 300nm to about 700μm, about 300nm to about 600μm, about 300nm to about 500μm, about 300nm to about 400μm, about 300nm to about 300μm, about 300nm to about 200μm, about 300nm to about 100μm, about 300nm to about 50μm, about 300nm to about 10μm , about 300nm to about 5μm, about 300nm to about 1μm, about 300nm to about 900nm, about 300nm to about 800nm, about 300nm to about 700nm, about 300nm to about 600nm, about 300nm to about 500nm, about 300nm to about 400nm, about 400nm to about 50mm, about 400nm to about 45mm, about 400nm to about 40mm, about 400nm to about 35mm, about 400nm to about 30mm, about 400nm to about 25mm, about 400nm to about 20mm, about 400nm to about 15mm, about 400nm to about 10mm, about 400nm to about 5mm, about 400nm to about 1m m, about 400nm to about 900μm, about 400nm to about 800μm, about 400nm to about 700μm, about 400nm to about 600μm, about 400nm to about 500μm, about 400nm to about 400μm, about 400nm to about 300μm, about 400nm to about 200μm, about 400nm to about 100μm, about 400nm to about 50μm, about 400nm to about 10μm, about 400nm to about 5μm, about 400nm to about 1μm, about 400nm to about 900nm, about 400nm to about 800nm, about 400nm to about 700nm, about 400nm to about 600nm, about 400nm to about 500nm,from about 500 nm to about 50 mm, from about 500 nm to about 45 mm, from about 500 nm to about 40 mm, from about 500 nm to about 35 mm, from about 500 nm to about 30 mm, from about 500 nm to about 25 mm, from about 500 nm to about 20 mm, from about 500 nm to about 15 mm, from about 500 nm to about 10 mm, from about 500 nm to about 5 mm, from about 500 nm to about 1 mm, from about 500 nm to about 900 μm, from about 500 nm to about 800 μm, from about 500 nm to about 700 μm, from about 500 nm to about 600 μm, from about 500 nm to about 500 μm, from about 500 nm to about 400 μm, from about 500 nm to about 300 μm, from about 500 nm to about 20 0 μm, about 500 nm to about 100 μm, about 500 nm to about 50 μm, about 500 nm to about 10 μm, about 500 nm to about 5 μm, about 500 nm to about 1 μm, about 500 nm to about 900 nm, about 500 nm to about 800 nm, about 500 nm to about 700 nm, about 500 nm to about 600 nm, about 600 nm to about 50 mm, about 600 nm to about 45 mm, about 600 nm to about 40 mm, about 600 nm to about 35 mm, about 600 nm to about 30 mm, about 600 nm to about 25 mm, about 600 nm to about 20 mm, about 600 nm to about 15 mm, about 600 nm to about 10 mm, about 600 nm to about 5 mm, about 600nm to about 1mm, about 600nm to about 900μm, about 600nm to about 800μm, about 600nm to about 700μm, about 600nm to about 600μm, about 600nm to about 500μm, about 600nm to about 400μm, about 600nm to about 300μm, about 600nm to about 200μm, about 600nm to about 100μm, about 600nm to about 50μm, about 600nm to about 10μm, about 600nm to about 5μm, about 600nm to about 1μm, about 600nm to about 900nm, about 600nm to about 800nm, about 600nm to about 700nm, about 700nm to about 50mm, about 700 700nm to about 45mm, about 700nm to about 40mm, about 700nm to about 35mm, about 700nm to about 30mm, about 700nm to about 25mm, about 700nm to about 20mm, about 700nm to about 15mm, about 700nm to about 10mm, about 700nm to about 5mm, about 700nm to about 1mm, about 700nm to about 900μm, about 700nm to about 800μm, about 700nm to about 700μm, about 700nm to about 600μm, about 700nm to about 500μm, about 700nm to about 400μm, about 700nm to about 300μm, about 700nm to about 200μm, about 700nm to about 100μm,about 700nm to about 50μm, about 700nm to about 10μm, about 700nm to about 5μm, about 700nm to about 1μm, about 700nm to about 900nm, about 700nm to about 800nm, about 800nm ​​to about 50mm, about 800nm ​​to about 45mm, about 800nm ​​to about 40mm, about 800nm ​​to about 35mm, about 800nm ​​to about 30mm, about 800nm ​​to about 25mm, about 800nm ​​to about 20mm, about 800nm ​​to about 15mm, about 800nm ​​to about 10mm, about 800nm ​​to about 5mm, about 800nm ​​to about 1mm, about 800nm ​​to about 900μm, about 800nm ​​to about 800μm, about 8 00nm to about 700μm, about 800nm ​​to about 600μm, about 800nm ​​to about 500μm, about 800nm ​​to about 400μm, about 800nm ​​to about 300μm, about 800nm ​​to about 200μm, about 800nm ​​to about 100μm, about 800nm ​​to about 50μm, about 800nm ​​to about 10μm, about 800nm ​​to about 5μm, about 800nm ​​to about 1μm, about 800nm ​​to about 900nm, about 900nm to about 50mm, about 900nm to about 45mm, about 900nm to about 40mm, about 900nm to about 35mm, about 900nm to about 30mm, about 900nm to about 25mm, about 900nm to about 20m m, about 900nm to about 15mm, about 900nm to about 10mm, about 900nm to about 5mm, about 900nm to about 1mm, about 900nm to about 900μm, about 900nm to about 800μm, about 900nm to about 700μm, about 900nm to about 600μm, about 900nm to about 500μm, about 900nm to about 400μm, about 900nm to about 300μm, about 900nm to about 200μm, about 900nm to about 100μm, about 900nm to about 50μm, about 900nm to about 10μm, about 900nm to about 5μm, about 900nm to about 1μm, about 1μm to about 50mm, about 1μm to about 45mm , about 1 μm to about 40 mm, about 1 μm to about 35 mm, about 1 μm to about 30 mm, about 1 μm to about 25 mm, about 1 μm to about 20 mm, about 1 μm to about 15 mm, about 1 μm to about 10 mm, about 1 μm to about 5 mm, about 1 μm to about 1 mm, about 1 μm to about 900 μm, about 1 μm to about 800 μm, about 1 μm to about 700 μm, about 1 μm to about 600 μm, about 1 μm to about 500 μm, about 1 μm to about 400 μm, about 1 μm to about 300 μm, about 1 μm to about 200 μm, about 1 μm to about 100 μm, about 1 μm to about 50 μm, about 1 μm to about 10 μm, about 1 μm to about 5 μm, about 5 μm to about 50 mm,from about 5 μm to about 45 mm, from about 5 μm to about 40 mm, from about 5 μm to about 35 mm, from about 5 μm to about 30 mm, from about 5 μm to about 25 mm, from about 5 μm to about 20 mm, from about 5 μm to about 15 mm, from about 5 μm to about 10 mm, from about 5 μm to about 5 mm, from about 5 μm to about 1 mm, from about 5 μm to about 900 μm, from about 5 μm to about 800 μm, from about 5 μm to about 700 μm, from about 5 μm to about 600 μm, from about 5 μm to about 500 μm, from about 5 μm to about 400 μm, from about 5 μm to about 300 μm, from about 5 μm to about 200 μm, from about 5 μm to about 100 μm, from about 5 μm to about 50 μm, from about 5 μm to about 10 μm, from about 10 μm to about 50 mm , about 10μm to about 45mm, about 10μm to about 40mm, about 10μm to about 35mm, about 10μm to about 30mm, about 10μm to about 25mm, about 10μm to about 20mm, about 10μm to about 15mm, about 10μm to about 10mm, about 10μm to about 5mm, about 10μm to about 1mm, about 10μm to about 900μm, about 10μm to about 800μm, about 10μm to about 700μm, about 10μm to about 600μm, about 10μm to about 500μm, about 10μm to about 400μm, about 10μm to about 300μm, about 10μm to about 200μm, about 10μm to about 100μm, about 10μm to about 50μm, about 50 μm to about 50 mm, about 50 μm to about 45 mm, about 50 μm to about 40 mm, about 50 μm to about 35 mm, about 50 μm to about 30 mm, about 50 μm to about 25 mm, about 50 μm to about 20 mm, about 50 μm to about 15 mm, about 50 μm to about 10 mm, about 50 μm to about 5 mm, about 50 μm to about 1 mm, about 50 μm to about 900 μm, about 50 μm to about 800 μm, about 50 μm to about 700 μm, about 50 μm to about 600 μm, about 50 μm to about 500 μm, about 50 μm to about 400 μm, about 50 μm to about 300 μm, about 50 μm to about 200 μm, about 50 μm to about 100 μm, about 10 0μm to about 50mm, about 100μm to about 45mm, about 100μm to about 40mm, about 100μm to about 35mm, about 100μm to about 30mm, about 100μm to about 25mm, about 100μm to about 20mm, about 100μm to about 15mm, about 100μm to about 10mm, about 100μm to about 5mm, about 100μm to about 1mm, about 100μm to about 900μm, about 100μm to about 800μm, about 100μm to about 700μm, about 100μm to about 600μm, about 100μm to about 500μm, about 100μm to about 400μm, about 100μm to about 300μm, about 100μm to about 200μm,from about 200 μm to about 50 mm, from about 200 μm to about 45 mm, from about 200 μm to about 40 mm, from about 200 μm to about 35 mm, from about 200 μm to about 30 mm, from about 200 μm to about 25 mm, from about 200 μm to about 20 mm, from about 200 μm to about 15 mm, from about 200 μm to about 10 mm, from about 200 μm to about 5 mm, from about 200 μm to about 1 mm, from about 200 μm to about 900 μm, from about 200 μm to about 800 μm, from about 200 μm to about 700 μm, from about 200 μm to about 600 μm, from about 200 μm to about 500 μm, from about 200 μm to about 400 μm, from about 200 μm to about 300 μm, from about 300 μm to about 50 mm m, about 300 μm to about 45 mm, about 300 μm to about 40 mm, about 300 μm to about 35 mm, about 300 μm to about 30 mm, about 300 μm to about 25 mm, about 300 μm to about 20 mm, about 300 μm to about 15 mm, about 300 μm to about 10 mm, about 300 μm to about 5 mm, about 300 μm to about 1 mm, about 300 μm to about 900 μm, about 300 μm to about 800 μm, about 300 μm to about 700 μm, about 300 μm to about 600 μm, about 300 μm to about 500 μm, about 300 μm to about 400 μm, about 400 μm to about 50 mm, about 400 μm to about 45 mm, about 400 μm to about 40 mm, about 400 μm to about 35 mm, about 400 μm to about 30 mm, about 400 μm to about 25 mm, about 400 μm to about 20 mm, about 400 μm to about 15 mm, about 400 μm to about 10 mm, about 400 μm to about 5 mm, about 400 μm to about 1 mm, about 400 μm to about 900 μm, about 400 μm to about 800 μm, about 400 μm to about 700 μm, about 400 μm to about 600 μm, about 400 μm to about 500 μm, about 500 μm to about 50 mm, about 500 μm to about 45 mm, about 500 μm to about 40 mm, about 500 μm to about 35 mm, about 500 μm to about 30 mm, about 500 μm to about 25 mm, about 500 μm to about 20 mm, about 500 μm to about 15 mm, about 500 μm to about 10 mm, about 500 μm to about 5 mm, about 500 μm to about 1 mm, about 500 μm to about 900 μm, about 500 μm to about 800 μm, about 500 μm to about 700 μm, about 500 μm to about 600 μm, about 600 μm to about 50 mm, about 600 μm to about 45 mm, about 600 μm to about 40 mm, about 600 μm to about 35 mm, about 600 μm to about 30 mm, about 600 μm to about 25 mm, about 600 μm to about 20 mm, about 600 μm to about 15 mm, about 600 μm to about 10 mm, about 600 μm to about 5 mm,From about 600 μm to about 1 mm, from about 600 μm to about 900 μm, from about 600 μm to about 800 μm, from about 600 μm to about 700 μm, from about 700 μm to about 50 mm, from about 700 μm to about 45 mm, from about 700 μm to about 40 mm, from about 700 μm to about 35 mm, from about 700 μm to about 30 mm, from about 700 μm to about 25 mm, from about 700 μm to about 20 mm, from about 700 μm to about 15 mm, from about 700 μm to about 10 mm, from about 700 μm to about 5 mm, from about 700 μm to about 1 mm, from about 700 μm to about 900 μm, from about 700 μm to about 800 μm, from about 800 μm to about 50 mm, from about 800 μm to about 45 mm m, about 800 μm to about 40 mm, about 800 μm to about 35 mm, about 800 μm to about 30 mm, about 800 μm to about 25 mm, about 800 μm to about 20 mm, about 800 μm to about 15 mm, about 800 μm to about 10 mm, about 800 μm to about 5 mm, about 800 μm to about 1 mm, about 800 μm to about 900 μm, about 900 μm to about 50 mm, about 900 μm to about 45 mm, about 900 μm to about 40 mm, about 900 μm to about 35 mm, about 900 μm to about 30 mm, about 900 μm to about 25 mm, about 900 μm to about 20 mm, about 900 μm to about 15 mm, about 900 μm to about 10 mm, about 900 μm to about 5 mm, about 900 μm to about 1 mm, about 1 mm to about 50 mm, about 1 mm to about 45 mm, about 1 mm to about 40 mm, about 1 mm to about 35 mm, about 1 mm to about 30 mm, about 1 mm to about 25 mm, about 1 mm to about 20 mm, about 1 mm to about 15 mm, about 1 mm to about 10 mm, about 1 mm to about 5 mm, about 5 mm to about 50 mm, about 5 mm to about 45 mm, about 5 mm to about 40 mm, about 5 mm to about 35 mm, about 5 mm to about 30 mm, about 5 mm to about 25 mm, about 5 mm to about 20 mm, about 5 mm to about 15 mm, about 5 mm to about 10 mm, about 10 mm to about 50 mm, about 10 mm m to about 45mm, about 10mm to about 40mm, about 10mm to about 35mm, about 10mm to about 30mm, about 10mm to about 25mm, about 10mm to about 20mm, about 10mm to about 15mm, about 15mm to about 50mm, about 15mm to about 45mm, about 15mm to about 40mm, about 15mm to about 35mm, about 15mm to about 30mm, about 15mm to about 25mm, about 15mm to about 20mm, about 20mm to about 50mm, about 20mm to about 45mm, about 20mm to about 40mm, about 20mm to about 35mm, about 20mm to about 30mm, about 20mm to about 25mm, about 25mm to about 50mm,From about 25 mm to about 45 mm, from about 25 mm to about 40 mm, from about 25 mm to about 35 mm, from about 25 mm to about 30 mm, from about 30 mm to about 50 mm, from about 30 mm to about 45 mm, from about 30 mm to about 40 mm, from about 30 mm to about 35 mm, from about 35 mm to about 50 mm, from about 35 mm to about 45 mm, from about 35 mm to about 40 mm, from about 40 mm to about 50 mm, from about 40 mm to about 45 mm, or from about 45 mm to about 50 mm.

[0299] In some embodiments, β-TCP is about 50% to about 70% porous, wherein the pores have an average pore size of about 50 μm to about 70 μm (eg, about 55 μm to about 65 μm or about 60 μm).

[0300] Compositions and kits

[0301] Also provided herein are compositions (e.g., pharmaceutical compositions) comprising any chimeric polypeptide or β-TCP binding polypeptide described herein. In some examples, the compositions may also include β-TCP (e.g., any type of β-TCP described herein). In some examples, β-TCP is formulated.

[0302] In some examples, β-TCP can be placed on or in a stent, mesh, or sponge (e.g., an absorbable sponge).

[0303] In some cases, a composition (e.g., a pharmaceutical composition) is placed in a sterile vial or preloaded syringe.

[0304] In some cases, the composition (e.g., pharmaceutical composition) is formulated for different routes of administration (e.g., intra-articularly, injected into a joint, or injected near a bone crack or fracture). Single or multiple administrations of any pharmaceutical composition described herein may be given to a subject, depending on, for example, the dose and frequency required and tolerated by the subject. The dosage of the pharmaceutical composition should provide a sufficient amount of the chimeric polypeptide to effectively treat or improve the condition (e.g., bone defect, fracture, cartilage defect, or cartilage loss) or symptom.

[0305] Also provided herein is a kit, which includes any chimeric polypeptide described herein or any β-TCP binding polypeptide or any composition described herein (e.g., pharmaceutical composition). In some embodiments, the kit may include instructions for performing any method described herein. In some cases, the kit may include at least one dose of any composition described herein (e.g., pharmaceutical composition). In some embodiments, the kit may include a syringe for administering any pharmaceutical composition described herein. The kit described herein is not limited thereto; other variations are apparent to those of ordinary skill in the art.

[0306] Method for preparing the composition

[0307] Also provided herein is a method for producing any of the compositions described herein. Any of the compositions provided herein can be produced using methods described herein or methods known in the art. For example, in order to produce a β-TCP scaffold, granulated β-TCP powder can be sintered, sieved and manufactured into a desired shape (e.g., any shape described herein). In some examples, the purity of β-TCP present in any of the compositions described herein can be greater than about 75%, greater than 80%, greater than 85%, greater than 90%, greater than 95% or greater than 99% purity. In some examples, the purity of β-TCP present in any of the compositions described herein can be greater than 1%, greater than 2%, greater than 3%, greater than 4%, greater than 5%, greater than 6%, greater than 7%, greater than 8%, greater than 9%, greater than 10%, greater than 12%, greater than 14%, greater than 15%, greater than 16%, greater than 18%, greater than 20%, greater than 22%, greater than 24%, greater than 25%, greater than 26%, greater than 28%, greater than 30%, greater than 31%, greater than 32%, greater than 33%, greater than 34%, greater than 35%, greater than 36%, greater than 37%, greater than 38%, greater than 39%, greater than 40%, greater than 41%, greater than 42%, greater than 43%, greater than 44%, greater than 45%, greater than 46%, greater than 47%, greater than 48%, greater than 49%, greater than 50%, greater than 51%, greater than 52%, greater than 53%, greater than 54%, greater than 55%, greater than 56%, greater than 57%, greater than 58%, greater than 59%, greater than 60%, greater than 61%, greater than 62%, greater than 63%, greater than 64%, greater than 65%, greater than 66%, greater than In some examples, β-TCP is prepared using similar methods, but as a complex with other agents, such as a biocompatible polymer, for example, polylactide-co-glycolide.

[0308] It will be apparent to those skilled in the art that the composition may further include one or more pore formers. Examples of pore formers include, for example, inorganic salts such as sodium chloride, sugars (e.g., sucrose or glucose), gelatin (e.g., gelatin spheres), or paraffin (e.g., paraffin spheres).

[0309] The compositions described herein can be produced by contacting any chimeric polypeptide or any β-TCP binding polypeptide with any type of β-TCP described herein. In some embodiments, β-TCP can be in the form of a granular / powder form, a porous form, a putty (e.g., a moldable putty), a paste, a stent, and / or a coating on a solid surface (e.g., a coating on a medical device).

[0310] Treatment

[0311] Provided herein are methods for promoting bone or cartilage formation in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any composition described herein. Some embodiments of these methods may also include first selecting a subject in need of bone or cartilage formation. In some embodiments, the composition is administered to the subject near a desired bone or cartilage formation site in the subject.

[0312] Also provided herein are methods for replacing and / or repairing bone or cartilage in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any composition described herein. Some embodiments of these methods may also include first selecting a subject in need of bone replacement, bone repair, cartilage replacement, or cartilage repair. In some embodiments, the composition is administered to the subject near a desired bone or cartilage replacement or repair site in the subject.

[0313] Also provided herein are methods for treating a fracture or bone loss in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any composition described herein. Some embodiments of these methods may also include first selecting a subject with a fracture or bone loss. In some embodiments, the composition is administered to the subject near the fracture or bone loss site in the subject.

[0314] Also provided herein are methods for repairing soft tissue in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any composition described herein. Some embodiments of these methods may also include first selecting a subject with a fracture or bone loss. In some embodiments, the composition is administered to the subject near the site of the fracture or bone loss in the subject.

[0315] Also provided herein are methods for locally delivering a therapeutic agent to a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of any composition described herein. Some embodiments of these methods may also include first selecting a subject with a fracture or bone loss. In some embodiments, the composition is administered to the subject near the site of the fracture or bone loss in the subject.

[0316] In some cases, the subject suffers from a bone fracture or bone defect.

[0317] In some cases, the subject requires fusion of the vertebrae of the spine.

[0318] In some cases, the subject has a cartilage tear or cartilage defect.

[0319] In other cases, the subject suffers from cartilage loss.

[0320] Methods for determining the efficacy of treatment of a bone fracture or bone loss in a subject are known in the art and include, for example, imaging techniques (eg, magnetic resonance imaging, X-ray, or computed tomography).

[0321] Methods of detecting bone or cartilage formation, or bone or cartilage replacement or repair in a subject are also known in the art and include, for example, imaging techniques (eg, magnetic resonance imaging, X-ray, or computed tomography).

[0322] Suitable animal models for treating fractures or bone loss, bone or cartilage formation, or bone or cartilage replacement or repair are known in the art. Non-limiting examples of such animal models are described in the Examples and, for example, Drosse et al., Tissue Engineering Part C 14(1):79-88, 2008; Histing et al., Bone 49:591-599, 2011; and Poser et al., Hindawi Publishing Corporation, BioMed Research International; Article ID 348635, 2014.

[0323] The present invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0324] The present invention provides embodiments including but not limited to the following:

[0325] 1. A chimeric polypeptide comprising:

[0326] (i) one or more β-tricalcium phosphate (βTCP)-binding sequences, wherein at least one of the one or more β-TCP-binding sequences is selected from the group consisting of: VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19);

[0327] as well as

[0328] (ii) Mammalian growth factors.

[0329] 2. The chimeric polypeptide of embodiment 1, wherein the chimeric polypeptide comprises two or more β-TCP-binding sequences.

[0330] 3. The chimeric polypeptide of embodiment 2, wherein each adjacent pair of said two or more β-TCP binding sequences are directly adjacent to each other.

[0331] 4. The chimeric polypeptide of embodiment 2, wherein each adjacent pair of the two or more β-TCP binding sequences is separated by a linker sequence.

[0332] 5. The chimeric polypeptide of embodiment 2, wherein the chimeric polypeptide comprises at least two different β-TCP-binding sequences.

[0333] 6. The chimeric polypeptide of embodiment 2, wherein the chimeric polypeptide comprises two or more copies of the same β-TCP-binding sequence.

[0334] 7. The chimeric polypeptide of embodiment 5, wherein at least one of the two or more β-TCP-binding sequences is LLADTTHHRPWT (SEQ ID NO: 1) or GQVLPTTTPSSP (SEQ ID NO: 19).

[0335] 8. The chimeric polypeptide of embodiment 5, wherein the at least two different β-TCP-binding sequences comprise LLADTTHHRPWT (SEQ ID NO: 1) and VIGESTHHRPWS (SEQ ID NO: 2).

[0336] 9. The chimeric polypeptide of embodiment 8, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTVIGESTHHRPWS (SEQ ID NO: 20).

[0337] 10. The chimeric polypeptide of embodiment 5, wherein the at least two different β-TCP-binding sequences comprise LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), and IIGESSHHKPFT (SEQ ID NO: 6).

[0338] 11. The chimeric polypeptide of embodiment 10, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFT (SEQ ID NO: 21).

[0339] 12. The chimeric polypeptide of embodiment 5, wherein the at least two different β-TCP-binding sequences comprise LLADTTHHRPWT (SEQ ID NO: 1), VIGESTHHRPWS (SEQ ID NO: 2), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), and ILAESTHHKPWT (SEQ ID NO: 4).

[0340] 13. The chimeric polypeptide of embodiment 12, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRP WGILAESTHHKPWT (SEQ ID NO: 22).

[0341] 14. The chimeric polypeptide of embodiment 5, wherein the at least two β-TCP-binding sequences comprise LLADTTHHRPWT (SEQ ID NO: 1) and ILAESTHHKPWT (SEQ ID NO: 4).

[0342] 15. The chimeric polypeptide of embodiment 14, wherein the chimeric polypeptide comprises at least one copy of the sequence LLADTTHHRPWTILAESTHHKPWT (SEQ ID NO: 23).

[0343] 16. The chimeric polypeptide of embodiment 15, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTILAESTHHKPWTLLADTTHHRPWTILAESTHHK PWTLLADTTHHRPWT (SEQ ID NO: 24).

[0344] 17. The chimeric polypeptide of embodiment 5, wherein the at least two β-TCP-binding sequences comprise LLADTTHHRPWT (SEQ ID NO: 1) and GLGDTTHHRPWG (SEQ ID NO: 7).

[0345] 18. The chimeric polypeptide of embodiment 17, wherein the chimeric polypeptide comprises at least one copy of the sequence of LLADTTHHRPWTGLGDTTHHRPWG (SEQ ID NO: 25).

[0346] 19. The chimeric polypeptide of embodiment 18, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 26).

[0347] 20. The chimeric polypeptide of embodiment 18, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHH RPWGLLADTTHHRPWT (SEQ ID NO: 27).

[0348] 21. The chimeric polypeptide of embodiment 18, wherein the chimeric polypeptide comprises the sequence of LLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWTGLGDTTHH RPWGLLADTTHHRPWTGLGDTTHHRPWGLLADTTHHRPWT (SEQ ID NO: 28).

[0349] 22. The chimeric polypeptide of embodiment 5, wherein the at least two β-TCP-binding sequences comprise STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), and GSSGESTHHKPST (SEQ ID NO: 14).

[0350] 23. The chimeric polypeptide of embodiment 22, wherein the chimeric polypeptide comprises the sequence of STADTSHHRPSTSGGESTHHRPSTSGGESSHHKPSTGSGDSSHHRPS GSSGESTHHKPST (SEQ ID NO: 29).

[0351] 24. The chimeric polypeptide of embodiment 5, wherein the at least two β-TCP-binding sequences comprise VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18), and GGADTTHHRPGT (SEQ ID NO: 19).

[0352] 25. The chimeric polypeptide of embodiment 24, wherein the chimeric polypeptide comprises the sequence of VGADSTHHRPVTGAADTTHHRPVTAGADTTHHRPVTGGADTTHHRPATGGADTTHHRPGT (SEQ ID NO: 30).

[0353] 26. The chimeric polypeptide of embodiment 5, wherein the at least two different β-TCP-binding sequences comprise STADTSHHRPS (SEQ ID NO: 10), LLADTTHHRPWT (SEQ ID NO: 1), TSGGESTHHRPS (SEQ ID NO: 11), VGADSTHHRPVT (SEQ ID NO: 15), TSGGESSHHKPS (SEQ ID NO: 12), GAADTTHHRPVT (SEQ ID NO: 16), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPS (SEQ ID NO: 14), and TGGADTTHHRPAT (SEQ ID NO: 18).

[0354] 27. The chimeric polypeptide of embodiment 26, wherein the chimeric polypeptide comprises the sequence of STADTSHHRPSLLADTTHHRPWTTSGGESTHHRPSVGADSTHHRP VTTSGGESSHHKPSGAADTTHHRPVTTGSGDSSHHRPSGSSGESTHHKPSTGGADTTHHRPAT (SEQ ID NO: 31).

[0355] 28. The chimeric polypeptide according to any one of embodiments 1-27,The mammalian growth factor is selected from the group consisting of epidermal growth factor (EGF), platelet-derived growth factor (PDGF), insulin-like growth factor (IGF-1), fibroblast growth factor (FGF), fibroblast growth factor 2 (FGF2), fibroblast growth factor 18 (FGF18), transforming growth factor α (TGF-α), transforming growth factor β (TGF-β), transforming growth factor β1 (TGF-β1), transforming growth factor β3 (TGF-β3), osteogenic protein 1 (OP-1), osteogenic protein 2 (OP-2), osteogenic protein 3 (OP-3), bone morphogenetic protein 2 (BMP-2), bone morphogenetic protein 3 (BMP-3), bone morphogenetic ... 4 (BMP-4), bone morphogenetic protein 5 (BMP-5), bone morphogenetic protein 6 (BMP-6), bone morphogenetic protein 7 (BMP-7), bone morphogenetic protein (BMP-9), bone morphogenetic protein 10 (BMP-10), bone morphogenetic protein 11 (BMP-11), bone morphogenetic protein 12 (BMP-12), bone morphogenetic protein protein 13 (BMP-13), bone morphogenetic protein 15 (BMP-15), dentin phosphoprotein (DPP), plant-related growth factor (Vgr), growth differentiation factor 1 (GDF-1), growth differentiation factor 3 (GDF-3), growth differentiation factor 5 (GDF-5), growth differentiation factor 6 (GDF-6), Growth differentiation factor 7 (GDF-7), growth differentiation factor 8 (GDF8), growth differentiation factor 11 (GDF11), growth differentiation factor 15 (GDF15), vascular endothelial growth factor (VEGF), hyaluronan binding protein (HABP), collagen binding protein (CBP), fibroblast growth factor 18 (FGF-18), keratinocyte growth factor (KGF), tumor necrosis factor α (TNFα), tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), wnt family member 1 (WNT1), wnt family member 2 (WNT2), wnt family member 2B (WNT2B), wnt family member 3 (WNT3), wnt family Wnt family member 3A (WNT3A), wnt family member 4 (WNT4), wnt family member 5A (WNT5A), wnt family member 5B (WNT5B), wnt family member 6 (WNT6), wnt family member 7A (WNT7A), wnt family member 7B (WNT7B), wnt family member 8A (WNT8A), wnt family member 8B (WNT8B), wnt family member 9A (WNT9A), wnt family member 9B (WNT9B), wnt family member 10A (WNT10A), wnt family member 10B (WNT10B), wnt family member 11 (WNT11) and wnt family member 16 (WNT16).

[0356] 29. The chimeric polypeptide of embodiment 28, wherein the mammalian growth factor is BMP-2.

[0357] 30. The chimeric polypeptide of embodiment 29, wherein the BMP-2 comprises MVAGTRCLLALLLPQVLLGGAAGLVPELGRRKFAAASSGRPSSQPSDEVLSEFELRLLSMFGLKQRPTPSRDAVVPPYMLDLYRRHSGQPGSPAPDHRLERAASRANTVRSFHHEESLEELPETSGKTTRRFFFNLSSIPTEEFITSAELQVFREQMQDALGNNSSFHHRINIYEIIKPATANSKF PVTRLLDTRLVNQNASRWESFDVTPAVMRWTAQGHANHGFVVEVAHLEEKQGVSKRHVRISRSLHQDEHSWSQIRPLLVTFGHDGKGHPLHKREKRQAKHKQRKRLKS SCKRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLADHLNSTNHAIVQTLVNSVNSKIPKACCVPTELSAISMLYLDENEKVVLKNYQDMVVEGCGCR(SEQID NO: 32) amino acid sequence.

[0358] 31. The chimeric polypeptide of embodiment 30, wherein the BMP-2 consists of the amino acids of SEQ ID NO: 32.

[0359] 32. The chimeric polypeptide of any one of embodiments 1-31, wherein the chimeric polypeptide further comprises a linker sequence between the adjacent pair of β-TCP-binding sequences and the growth factor.

[0360] 33. The chimeric polypeptide of embodiment 32, wherein the linker sequence comprises TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33).

[0361] 34. The chimeric polypeptide of embodiment 33, wherein the linker sequence consists of SEQ ID NO: 33.

[0362] 35. The chimeric polypeptide of embodiment 32, wherein the linker sequence comprises GAGTG (SEQ ID NO: 34).

[0363] 36. The chimeric polypeptide of embodiment 35, wherein the linker sequence consists of SEQ ID NO: 34.

[0364] 37. The chimeric polypeptide of any one of embodiments 1-36, wherein the chimeric polypeptide has a K of about 1 picomolar to about 100 micromolar. D Binds to β-TCP.

[0365] 38. A composition comprising the chimeric polypeptide of any one of embodiments 1-37.

[0366] 39. The composition of embodiment 38, wherein the composition further comprises β-TCP.

[0367] 40. The composition of embodiment 39, wherein the β-TCP is formulated as a powder, putty, or paste.

[0368] 41. The composition of embodiment 39, wherein the β-TCP is disposed on or in a scaffold, mesh, or sponge.

[0369] 42. A composition according to any one of embodiments 38-41, wherein the composition is a pharmaceutical composition.

[0370] 43. A kit comprising the composition according to any one of embodiments 38-42.

[0371] 44. A chimeric polypeptide comprising:

[0372] (i) one or more β-TCP-binding sequences comprising the amino acid sequence of Formula I:

[0373] A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein:

[0374] A0 is V, L, I, G, S, T, or A;

[0375] B0 is I, L, V, Q, T, S, G, or A;

[0376] C0 is G, A, V or S;

[0377] D0 is E, D, L or G;

[0378] E0 is S, T, P, T, E, or D;

[0379] F0 is T or S;

[0380] G0 is H, T or S;

[0381] H0 is H or T;

[0382] I0 is R, S, K, P or H;

[0383] J0 is P, S, R or K;

[0384] K0 is W, F, S, P, V, A, or G; and

[0385] L0 does not exist or is S, T, G (or A); and

[0386] (ii) mammalian growth factors,

[0387] Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0388] 45. The chimeric polypeptide of embodiment 44, wherein the chimeric polypeptide comprises two or more β-TCP-binding sequences of Formula I.

[0389] 46. ​​The chimeric polypeptide of embodiment 45, wherein each adjacent pair of said two or more β-TCP binding sequences of Formula I are directly adjacent to each other.

[0390] 47. The chimeric polypeptide of embodiment 45, wherein each adjacent pair of said two or more β-TCP binding sequences of Formula I are separated by a linker sequence.

[0391] 48. The chimeric polypeptide of embodiment 45, wherein the chimeric polypeptide comprises at least two different β-TCP-binding sequences of Formula I.

[0392] 49. The chimeric polypeptide of embodiment 45, wherein the chimeric polypeptide comprises two or more copies of the same β-TCP-binding sequence of Formula I.

[0393] 50. The chimeric polypeptide of embodiment 44, wherein the chimeric polypeptide further comprises a β-TCP-binding sequence that does not include the sequence of Formula I.

[0394] 51. The chimeric polypeptide of any one of embodiments 44-50, wherein the mammalian growth factor is selected from the group consisting of: EGF, PDGF, IGF-1, FGF, FGF2, FGF18, TGF-α, TGF-β, TGF-β1, TGF-β3, OP-1, OP-2, OP-3, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-9, BMP-10, BMP-12, BMP-11, BMP-13, BMP-15, DPP, Vg r, GDF-1, GDF-3, GDF-5, GDF-6, GDF-7, GDF8, GDF11, GDF15, VEGF, HABP, CBP, FGF-18, KGF, TNFα, TRAIL, WNT1, WNT2, WNT 2B, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, and WNT16.

[0395] 52. The chimeric polypeptide of embodiment 51, wherein the mammalian growth factor is BMP-2.

[0396] 53. The chimeric polypeptide of embodiment 52, wherein the BMP-2 comprises MVAGTRCLLALLLPQVLLGGAAGLVPELGRRKFAAASSGRPSSQPSDEVLSEFELRLLSMFGLKQRPTPSRDAVVPPYMLDLYRRHSGQPGSPAPDHRLERAASRANTVRSFHHEESLEELPETSGKTTRRFFFNLSSIPTEEFITSAELQVFREQMQDALGNNSSFHHRINIYEIIKPATANSKF PVTRLLDTRLVNQNASRWESFDVTPAVMRWTAQGHANHGFVVEVAHLEEKQGVSKRHVRISRSLHQDEHSWSQIRPLLVTFGHDGKGHPLHKREKRQAKHKQRKRLKS SCKRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLADHLNSTNHAIVQTLVNSVNSKIPKACCVPTELSAISMLYLDENEKVVLKNYQDMVVEGCGCR(SEQID NO: 32) amino acid sequence.

[0397] 54. The chimeric polypeptide of embodiment 53, wherein the BMP-2 consists of the amino acids of SEQ ID NO: 32.

[0398] 55. The chimeric polypeptide of any one of embodiments 44-54, wherein the chimeric polypeptide further comprises a linker sequence between the adjacent pair of β-TCP-binding sequences and the growth factor.

[0399] 56. The chimeric polypeptide of embodiment 55, wherein the linker sequence comprises TGGSGEGGTGASTGGSAGTGGSGGTTSGEAGGSSGAG (SEQ ID NO: 33).

[0400] 57. The chimeric polypeptide of embodiment 56, wherein the linker sequence consists of SEQ ID NO: 33.

[0401] 58. The chimeric polypeptide of embodiment 55, wherein the linker sequence comprises GSGATG (SEQ ID NO: 34).

[0402] 59. The chimeric polypeptide of embodiment 58, wherein the linker sequence consists of SEQ ID NO: 34.

[0403] 60. The chimeric polypeptide of any one of embodiments 44-59, wherein the chimeric polypeptide has a K of about 1 picomolar to about 100 micromolar. D Binds to β-TCP.

[0404] 61. A composition comprising the chimeric polypeptide of any one of embodiments 44-60.

[0405] 62. The composition of embodiment 61, wherein the composition further comprises β-TCP.

[0406] 63. The composition of embodiment 62, wherein the β-TCP is formulated as a powder, putty, or paste.

[0407] 64. The composition of embodiment 62, wherein the β-TCP is disposed on or in a scaffold, mesh, or sponge.

[0408] 65. A composition according to any one of embodiments 61-64, wherein the composition is a pharmaceutical composition.

[0409] 66. A kit comprising the composition according to any one of embodiments 61-65.

[0410] 67. A method of promoting bone or cartilage formation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to any one of embodiments 38-42 and 61-65.

[0411] 68. A method of replacing and / or repairing bone or cartilage in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to any one of embodiments 38-42 and 61-65.

[0412] 69. A method of treating bone fracture or bone loss in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to any one of embodiments 38-42 and 61-65.

[0413] 70. A method according to any one of embodiments 67-69, wherein the subject suffers from a bone fracture.

[0414] 71. The method of any one of embodiments 67-69, wherein the subject has a bone defect.

[0415] 72. The method of embodiment 67 or 68, wherein the subject suffers from a cartilage tear or a cartilage defect.

[0416] 73. A method according to embodiment 67 or 68, wherein the subject suffers from cartilage loss.

[0417] 74. A beta-tricalcium phosphate-binding polypeptide comprising at least one sequence selected from the group consisting of VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17). NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19).

[0418] 75. A β-tricalcium phosphate-binding polypeptide comprising the amino acid sequence of formula I:

[0419] A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein:

[0420] A0 is V, L, I, G, S, T, or A;

[0421] B0 is I, L, V, Q, T, S, G, or A;

[0422] C0 is G, A, V or S;

[0423] D0 is E, D, L or G;

[0424] E0 is S, T, P, T, E, or D;

[0425] F0 is T or S;

[0426] G0 is H, T or S;

[0427] H0 is H or T;

[0428] I0 is R, S, K, P or H;

[0429] J0 is P, S, R or K;

[0430] K0 is W, F, S, P, V, A, or G; and

[0431] L0 does not exist or is S, T, G (or A);

[0432] Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

[0433] Example

[0434] Example 1. Preclinical study of tBMP-2 in a β-TCP carrier in a rat critical-size femoral defect model

[0435] The purpose of this study is to test the safety and effect of modified recombinant human bone morphogenetic protein (rhBMP) (referred to as tBMP) in a β-tricalcium phosphate (β-TCP) putty matrix in bone replacement and repair in a rat critical size defect model. The tBMP-2 variant exhibits a very tight binding to calcium phosphate ceramics, thus allowing tBMP-2 to be delivered to the implant site in a targeted manner. This is achieved by surgically creating a defect using the RatFix internal fixator system, and then filling the defect with tBMP+β-TCP putty or only β-TCP putty as a control. The positive control is a commercially available (Medtronic) formulation of rhBMP with an absorbable collagen sponge (ACS) for human use. Animals are monitored clinically and radiographed every two weeks until their predetermined endpoints of four weeks or eight weeks after the treatment is administered. Organs and tissues are examined ex vivo histologically, and μCT, tissue morphology, and / or mechanical strength tests are performed on the treated legs.

[0436] This report covers the arrival of the animals, grouping, surgically created defects, and administration of the test, reference, or control items. In addition, this report covers the postoperative period up to and including week 8, as well as endpoints measured in vivo and in vitro. Included are clinical summaries, radiographs, endpoint clinical pathology and organ histopathology, histomorphometry of the treated leg (odd-numbered animals), and mechanical strength testing of the treated leg (even-numbered animals).

[0437] Upon arrival at the testing facility, all animals received a veterinary health check and were weighed. Based on body weight, animals were divided into groups A): tBMP + β-TCP putty; B) rhBMP + ACS; or C) β-TCP putty only. The animals were then given at least five days to acclimate to their new environment and diet.

[0438] On the day of surgery, the animal is anesthetized, the defect is created, and the bone is supported with an internal fixator according to the manufacturer's protocol (with some refinements to the surgical procedure). The pre-prepared test, reference, or control item is then inserted in...

Claims

1. A chimeric polypeptide comprising: (i) one or more β-tricalcium phosphate (βTCP)-binding sequences, wherein at least one of the one or more β-TCP-binding sequences is selected from the group consisting of: VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), NO: 16), AGADTTHHRPVT (SEQ ID NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19); as well as (ii) Mammalian growth factors.

2. A composition comprising the chimeric polypeptide according to claim 1.

3. A kit comprising the composition according to claim 2.

4. A chimeric polypeptide comprising: (i) one or more β-TCP-binding sequences comprising the amino acid sequence of Formula I: A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein: A0 is V, L, I, G, S, T, or A; B0 is I, L, V, Q, T, S, G, or A; C0 is G, A, V or S; D0 is E, D, L or G; E0 is S, T, P, T, E, or D; F0 is T or S; G0 is H, T or S; H0 is H or T; I0 is R, S, K, P or H; J0 is P, S, R or K; K0 is W, F, S, P, V, A, or G; and L0 does not exist or is S, T, G (or A); and (ii) mammalian growth factors, Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

5. A composition comprising the chimeric polypeptide according to claim 4.

6. A method of promoting bone or cartilage formation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the composition according to claim 2 or 5.

7. A method of replacing and / or repairing bone or cartilage in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition according to claim 2 or 5.

8. A method of treating bone fracture or bone loss in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the composition of claim 2 or 5.

9. A β-tricalcium phosphate-binding polypeptide comprising at least one sequence selected from the group consisting of VIGESTHHRPWS (SEQ ID NO: 2), LIADSTHHSPWT (SEQ ID NO: 3), ILAESTHHKPWT (SEQ ID NO: 4), ILAETTHHRPWS (SEQ ID NO: 5), IIGESSHHKPFT (SEQ ID NO: 6), GLGDTTHHRPWG (SEQ ID NO: 7), VLGDTTHHKPWT (SEQ ID NO: 8), IVADSTHHRPWT (SEQ ID NO: 9); STADTSHHRPS (SEQ ID NO: 10), TSGGESTHHRPS (SEQ ID NO: 11), TSGGESSHHKPS (SEQ ID NO: 12), TGSGDSSHHRPS (SEQ ID NO: 13), GSSGESTHHKPST (SEQ ID NO: 14), VGADSTHHRPVT (SEQ ID NO: 15), GAADTTHHRPVT (SEQ ID NO: 16), AGADTTHHRPVT (SEQ ID NO: 17). NO: 17), GGADTTHHRPAT (SEQ ID NO: 18) and GGADTTHHRPGT (SEQ ID NO: 19).

10. A β-tricalcium phosphate-binding polypeptide comprising the amino acid sequence of formula I: A0B0C0D0E0F0G0H0I0J0K0L0 (Formula I) (SEQ ID NO: 35), wherein: A0 is V, L, I, G, S, T, or A; B0 is I, L, V, Q, T, S, G, or A; C0 is G, A, V or S; D0 is E, D, L or G; E0 is S, T, P, T, E, or D; F0 is T or S; G0 is H, T or S; H0 is H or T; I0 is R, S, K, P or H; J0 is P, S, R or K; K0 is W, F, S, P, V, A, or G; and L0 does not exist or is S, T, G (or A); Wherein Formula I does not include LLADTTHHRPWT (SEQ ID NO: 1).

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