Antibody-peptide fusion proteins for treating amyloid disorders

Abstract

Provided herein are amyloid-reactive peptides and antibody-peptide fusion proteins. Also provided herein are methods of treating amyloid-based diseases by administering an antibody-peptide fusion protein.

Description

Attorney Docket No. 165992001640ANTIBOD Y-PEPTIDE FUSION PROTEINS FOR TREATING AMYLOID DISORDERSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 691,858 filed September 6, 2024, hereby incorporated by reference in its entirety.INCORPORATION BY REFERENCE OF SEUQENCE LISTING

[0002] This present application is being filed along with a Sequence Listing in electronic format. The content of the electronic sequence listing (165992001640seqlist.xml; Size: 88,823 bytes; and Date of Creation: September 2, 2025) is herein incorporated by reference in its entirety.FIELD OF THE INVENTION

[0003] This application relates to amyloid-reactive peptides, antibody-peptide fusion proteins that bind to human amyloid fibrils, and methods of using the same.BACKGROUND

[0004] Systemic amyloidosis is characterized by the deposition of misfolded proteins, as amyloid fibrils, in the extracellular space of organs and tissues throughout the body, Merlini, G., Systemic amyloidosis: are we moving aheadd Neth J Med, 2004. 62(4): p. 104-5; Gertz, M.A. and A. Dispenzieri, Systemic Amyloidosis Recognition, Prognosis, and Therapy: A Systematic Review. JAMA, 2020. 324(1): p. 79-89. The major types of systemic amyloidosis, ATTR and AL, result from the deposition of transthyretin and monoclonal immunoglobulin light chains, respectively, Muchtar, E., et al., Systemic amyloidosis from A (AA) to T (ATTR): a review. J Intern Med, 2021. 289(3): p. 268-292. Amyloid deposits are composed of amyloid fibrils in association with numerous accessory molecules, notably extracellular matrix components such as heparan sulfate proteoglycans and serum proteins sequestered from the circulation, Vrana, J. A., et al., Classification of amyloidosis by laser microdissection and mass spectrometry-based proteomic analysis in clinical biopsy specimens. Blood, 2009.114(24): p. 4957-9. Despite the presence of misfolded, non-native proteins in the amyloid fibrils, amyloid deposits are immunologically inert and not recognized or cleared by the immune system.

[0005] In addition to the inability to clear tissue amyloid, the clinical manifestations are many and diverse, which makes early and accurate diagnosis challenging, McCausland, K.L.,1MOFO-360384346.1Attorney Docket No. 165992001640 et al., Light Chain (AL) Amyloidosis: The Journey to Diagnosis. Patient, 2018. 11(2): p. 207- 216. The major causes of mortality are cardiac or renal failure. Cardiac amyloidosis can manifest as left ventricular wall and intraventricular septal hypertrophy and functional abnormalities that result in impaired global longitudinal strain and reduced ejection volume, Rubin, J. and M.S. Maurer, Cardiac Amyloidosis: Overlooked, Underappreciated, and Treatable. Annu Rev Med, 2020. 71 : p. 203-219. Amyloid-related renal insufficiency, a consequence of tubular and glomerular nephropathy, leads to proteinuria and worsening glomerular filtration, which ultimately results in the need for dialysis. The positive correlation between increasing amyloid load and worsening clinical outcomes has been well established, based on biopsy studies and measurements of extracellular volume studies of the kidney and heart, respectively, Banypersad, S.M., et al., T1 mapping and survival in systemic light-chain amyloidosis. Eur Heart J, 2015. 36(4): p. 244-51; Kristen, A.V., et al., Cardiac Amyloid Load: A Prognostic and Predictive Biomarker in Patients With Light-Chain Amyloidosis. J Am Coll Cardiol, 2016. 68(1): p. 13-2; Kuroda, T., et al., Significant association between renal function and amyloid-positive area in renal biopsy specimens in AL amyloidosis. BMC Nephrol, 2012. 13: p. 118.

[0006] Current treatment strategies focus on reducing the availability of the amyloidogenic precursor protein, thereby slowing amyloid formation. For patients with AL amyloidosis, this is primarily achieved with plasma cell-directed chemo- and immunotherapeutics, protease inhibitors, and autologous stem cell transplantation, Al Hamed, R., et al., Comprehensive Review of AL amyloidosis: some practical recommendations. Blood Cancer J, 2021. 11(5): p. 97; Dima, D., et al., Diagnostic and Treatment Strategies for AL Amyloidosis in an Era of Therapeutic Innovation. JCO Oncology Practice, 2023. 19(5): p. 265-275. In patients with ATTR, transthyretin production by the liver is reduced using ribonucleic acid interference (RNAi), Adams, D., et al., Patisiran, an RNAi Therapeutic, for Hereditary Transthyretin Amyloidosis. N Engl J Med, 2018. 379(1): p. 11-21; Adams, D., et al., Long-term safety and efficacy of patisiran for hereditary transthyretin-mediated amyloidosis with polyneuropathy: 12-month results of an open-label extension study. Lancet Neurol, 2021. 20(1): p. 49-59, or ligand-conjugated antisense oligonucleotides, Benson, M.D., et al., Inotersen Treatment for Patients with Hereditary Transthyretin Amyloidosis. N Engl J Med, 2018. 379(1): p. 22-31. Additionally, for patients with ATTR amyloidosis, stabilization of the tetrameric transthyretin native state is achieved using small molecule ligands that prevent dissociation, which is a critical first step in amyloid formation, Coelho, T., et al., Design and Rationale of the Global Phase 3 NEURO -TTRansf or m Study of2MOFO-360384346.1Attorney Docket No. 165992001640Antisense Oligonucleotide AKCEA-TTR-L(Rx) (ION-682884-CS3) in Hereditary Transthyretin-Mediated Amyloid Polyneuropathy. Neurol Ther, 2021. 10(1): p. 375-38; Hammarstrom, P., et al., Prevention of transthyretin amyloid disease by changing protein misfolding energetics. Science, 2003. 299(5607): p. 713-6; Maurer, M.S., et al., Tafamidis Treatment for Patients with Transthyretin Amyloid Cardiomyopathy. N Engl J Med, 2018. 379(11): p. 1007-1016; Penchala, S.C., et al., AGIO inhibits amyloidogenesis and cellular toxicity of the familial amyloid cardiomyopathy-associated V122I transthyretin. Proc Natl Acad Sci U S A, 2013. 110(24): p. 9992-7. At present, there are no approved methods to clear amyloid from organs, and the prognosis often remains poor for these patients.Moreover, for patients with the many rare forms of amyloidosis, there are no therapeutic options. Therefore, novel reagents capable of clearing amyloid and thereby restoring organ dysfunction represent an important goal in the treatment of these disorders.

[0007] Opsonization of amyloid with immunostimulatory antibodies and antibody fragments that can induce recruitment and stimulation of macrophages to clear tissue amyloid is a novel approach to effect amyloid clearance and potentially improve patient outcomes, Bodin, K., et al., Antibodies to human serum amyloid P component eliminate visceral amyloid deposits. Nature, 2010. 468(7320): p. 93-7; Hmcic, R., et al., Antibody-mediated resolution of light chain-associated amyloid deposits. Am J Pathol, 2000. 157(4): p. 1239-46; Mi chai on, A., et al., A human antibody selective for transthyretin amyloid removes cardiac amyloid through phagocytic immune cells. Nat Commun, 2021. 12(1): p. 3142.

[0008] Accordingly, there is a need for effective treatments for amyloidosis and amyloid related diseases.SUMMARY OF THE INVENTION

[0009] Provided herein are antibody-peptide fusion proteins comprising an amyloidreactive peptide linked to an antibody, as well as methods of making and using the like.

[0010] Provided herein are amyloid-reactive peptides that binds to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K wherein n is at least 2, wherein the amyloid reactive-peptide binds to human amyloid fibrils with a Ec50 that is less than about 1000 nM. In some aspects, x is selected from a group consisting of A and G. In some aspects, n is between 2 and 20. In some aspects, y is R. In some aspects, y is K.

[0011] Also provided herein are amyloid-reactive peptides that binds to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amino acid sequence selected from3MOFO-360384346.1Attorney Docket No. 165992001640 the group consisting of SEQ ID NOs: 1-8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions.

[0012] In some aspects, the amyloid reactive-peptide binds to human amyloid fibrils with a Ec50 that is less than about 1000 nM. In some aspects, the amino acid substitutions are conservative substitutions.

[0013] In some aspects, the amyloid-reactive peptide binds to rVX6WIL fibrils, ATTRwt(PER125) extract, ATTRv(KEN) extract, ALX(SHI) liver extract, ALK(TAL) liver extract, AP(l-40), IAAP, ALK4, ALX1, or ATTR fibrils. In some aspects, the amyloidreactive peptide binds to A LX, ALK, ATTRV, or rVX6WIL fibrils.

[0014] Also provided herein are antibody-peptide fusion proteins, comprising: the amyloid-reactive peptides provided herein joined to an antibody or fragment thereof. In some aspects, the antibody or fragment thereof binds to human amyloid. In some aspects, the antibody or fragment thereof binds to human immunoglobulin light chain amyloid. In some aspects, the antibody is a full-length antibody, a Fab fragment, or a scFv.

[0015] In some aspects, the antibody or fragment thereof comprises a human Fc region. In some aspects, the human Fc region is of an IgGl, IgG2, IgG3, or IgG4 isotype.

[0016] In some aspects, the antibody or fragment thereof comprises a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), wherein: a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19; b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19; or c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH4MOFO-360384346.1Attorney Docket No. 165992001640 comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19.

[0017] In some aspects, the amyloid-reactive peptide and antibody or fragment thereof are joined together at the N-terminal end of the light chain, the C-terminal end of the light chain, the N-terminal end of the heavy chain and / or the C-terminal end of the heavy chain. In some aspects, the amyloid-reactive peptide and antibody or fragment thereof are joined via a linker. In some aspects, the linker comprises an amino acid sequence set forth in SEQ ID NO 82.

[0018] In some aspects, the amyloid-reactive peptide and antibody or fragment thereof comprise an N-terminal spacer. In some aspects, the N-terminal spacer comprises an amino acid sequence set forth in SEQ ID NO: 83 or SEQ ID NO: 103.

[0019] In some aspects, the amyloid-reactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and the antibody-peptide fusion protein comprises a secretory leader sequence. In some aspects, the secretory leader sequence comprises an amino acid sequence set forth in SEQ ID NO: 84.

[0020] In some aspects, the antibody or functional fragment thereof comprises human framework sequences. In some aspects, the antibody is humanized. In some aspects, the VL comprises one or more amino acid residues selected from the group consisting of: a. Tyr at position 36; b. Leu at position 37; c. Leu at position 46; d. Leu at position 85; and e. Phe at position 87, wherein the VH comprises one or more amino acid residues selected from the group consisting of: a. Vai at position 37; b. Leu at position 48; c. Leu at position 67; d. Ser at position 68; e. Lys at position 71; f. Ser at position 76; g. Vai at position 78; h. Leu at position 79; i. Phe at position 80; j. Thr at position 89; k. Vai at position 93; and 1. Thr at position 94, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some aspects, the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises Leu at position 48, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some aspects, the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19.5MOFO-360384346.1Attorney Docket No. 165992001640

[0021] In some aspects, the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32-42. In some aspects, the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63. In some aspects, the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO:55. In some aspects, the VL comprises an amino acid sequence set forth in SEQ ID NO:34, and the VH comprises an amino acid sequence set forth in SEQ ID NO:48.

[0022] In some aspects, the amyloid-reactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and wherein the light chain with the amyloid-reactive peptide fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 86, 87, and 93-97.

[0023] In some aspects, the light chain with the amyloid-reactive peptide fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NO: 93 or 86.

[0024] In some aspects, the amyloid-reactive peptide and antibody or fragment thereof are joined together at the N-terminal end of the light chain and wherein the light chain with the amyloid-reactive peptide fused to the N- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 85, 89, 91, and 92.

[0025] In some aspects, the amyloid-reactive peptide is conjugated to a detectable label. In some aspects, the detectable label is selected from the group consisting ofnC,18F,i nIn, "mTc,89Ga,123I,124I,125I, a biotin and a fluorophore. In some aspects, the detectable label is124I. In some aspects, the detectable label is "mTc.

[0026] Also provided herein are pharmaceutical compositions comprising an amyloid reactive peptide provided herein or an amyloid-peptide fusion protein described herein. In some aspects, the pharmaceutical composition comprises a pharmaceutical acceptable carrier. In some aspects, the pharmaceutical composition is formulated for intravenous or subcutaneous administration.

[0027] Also provided herein are nucleic acid(s) encoding an amyloid reactive peptide described herein or an amyloid-peptide fusion protein described herein. Also provided herein are vectors comprising the nucleic acid(s). Also provided herein are host cells comprising the vector. In some aspects, the host cell is a mammalian cell, optionally a Chinese hamster ovary (CHO) cell.

[0028] Also provided herein are methods of making an antibody-peptide fusion comprising culturing a host cell described herein under conditions suitable for expression of6MOFO-360384346.1Attorney Docket No. 165992001640 the vector encoding an antibody -peptide fusion protein and recovering the antibody -peptide fusion protein.

[0029] Also provided herein are methods of treating a subject having an amyloid-related disorder comprising an amyloid deposit, comprising administering to the subject an effective amount of an antibody-peptide fusion protein described herein.

[0030] In some aspects, treating the subject with the antibody-peptide fusion protein causes phagocytosis of the amyloid deposit, causes clearance of the amyloid deposit and or prevents formation of additional amyloid deposits.

[0031] Also provided herein are methods of identifying an amyloid deposit in a subject, comprising administering an amyloid-reactive peptide or an antibody-peptide fusion protein described herein to the subject, and detecting a signal from the detectable label. In some aspects, the methods further comprising diagnosing the subject with amyloid-related disorder if a signal is detected. In some aspects, the methods further comprising treating the subject for an amyloid-related disorder if a signal is detected.

[0032] In some aspects, the amyloid-related disorder is systemic amyloidosis or localized amyloidosis. In some aspects, the amyloid-related disorder is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, APrP, and Ap amyloidosis. In some aspects, the amyloid-related disorder is systemic amyloidosis. In some aspects, the amyloid- related disorder is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, and ACys, In some aspects, the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

[0033] In some aspects, the amyloid deposit is opsonized by the antibody-peptide fusion protein.

[0034] In some aspects, the subject is a human.BRIEF DESCRIPTION OF THE DRAWINGS

[0035] FIG. 1A-1C show circular dichroism (CD) spectra of peptides. FIG. 1A shows a CD spectrum of pl 12 titrated with enoxaparin. FIG. IB shows a CD spectrum of p5+14 titrated with enoxaparin. FIG. 1C shows an exemplary CD spectrum for a typical alphahelical protein bovine serum albumin (BSA).7MOFO-360384346.1Attorney Docket No. 165992001640

[0036] FIG. 2 shows percent of radioiodinated pl 12 and p5+14 binding to synthetic amyloid-like fibrils composed of a recombinant X6 light chain variable domain (rVX6WIL fibrils).

[0037] FIG. 3A and 3B show binding of biotinylated amyloid-reactive peptides to human AL, human ATTR amyloid extracts and synthetic fibrils bound on a microplate. FIG. 3A shows binding of pl 09 (using SuperBlock [SB] blocking agent) and FIG. 3B shows binding of pl 10 to the same substrates.

[0038] FIG. 4A and 4B show binding of APF4 and APF2 to synthetic amyloid fibrils on a microplate. FIG. 4A shows binding to rVk6WIL fibrils and FIG. 4B shows binding to human ALA extract.

[0039] FIG. 5A-5C shows microautoradiography (ARG) images (5x and lOx) of radio- labeled peptides colocalized with amyloid deposits in the heart, liver, spleen, and kidney of mice with systemic serum amyloid A-associated (AA) amyloidosis. FIG. 5A shows selective binding of125I-plO8 to amyloid. FIG. 5B shows selective binding of125I-plO9 to amyloid. FIG. 5C shows selective binding of125I-p 110 to amyloid.

[0040] FIG 6A- 6B show biodistribution of125I-labeled pl 12 peptides in mice with systemic AA amyloidosis and wild type mice measured as the percent of the injected dose per gram of tissue (%ID / g) detectable 1 hour (FIG. 6A) and 4 hours (FIG. 6B) after injection. As used in FIG. 6A and FIG. 6B, ID represents injected dose.

[0041] FIG. 7A-7J show immunohistochemistry (IHC) slides of biotinylated peptides p5, pl 09, and pl 10 binding to human AL and ATTRv tissue amyloid as well as normal tissue. In FIG. 9A-H Congo red staining of the tissue is used to show the presence of amyloid deposits. FIG. 7A shows binding to ALX(BAB) amyloid in heart tissue. FIG. 7B shows binding to ALX(BAB) amyloid in liver tissue. FIG. 7C shows binding to ATTRv(GRE) amyloid in heart tissue. FIG. 7D shows binding to ALK(LAU) amyloid in kidney tissue. FIG. 7E shows binding to ALL(SHI) amyloid in liver tissue. FIG. 7F shows binding to ALX(SHI) amyloid in spleen tissue. FIG. 7G shows binding to ALK(TAL) amyloid in heart tissue. FIG. 7H shows binding to ALK(TAL) amyloid in kidney tissue. FIG. 71 shows a lack of peptide binding to normal heart tissue. FIG. 71 shows binding in normal kidney tissue.

[0042] FIG. 8 shows antibody-peptide fusion 1 (APF1), APF2, and APF4, binding to heparin at varied concentrations.

[0043] FIG. 9 shows antibody-peptide fusion 1 (APF1), APF8, and APF6, binding to enoxaparin-coated microplates at varied concentrations.8MOFO-360384346.1Attorney Docket No. 165992001640

[0044] FIG. 10 shows pHRodo red fluorescence emission levels as a measure of phagocytosis of pHRodo red-labeled rV / LWIL fibrils and human ALX(BAL) amyloid extract by human THP-1 cells in the presence of human control (ctrl) IgGl-Fc and IgGl antibodies and antibody-peptide fusion proteins.

[0045] FIG. 11 shows pHRodo red fluorescence levels as a measure of phagocytosis of pHRodo red-labeled rV / LWIL fibrils and ALX(BAL) by human THP-1 cells in the presence of in the presence of a control (ctrl) human IgGl antibody and APF2.

[0046] FIG. 12A and 12B show schematic representations of antibody-peptide fusion proteins. FIG. 12A shows a schematic representation of an antibody-peptide fusion protein wherein the amyloid-reactive peptides are conjugated to each N-termini of the antibody light chain protein. FIG. 12B shows a schematic representation of an antibody-peptide fusion protein wherein the amyloid-reactive peptides are conjugated to the C-termini of the antibody light chain protein.

[0047] FIG 13 shows measured bioactivity of antibody-peptide fusion proteins measured as binding to a heparin plate following incubation in mouse serum at 37°C for 0 to 80 hours. The percent of bioactivity at each time point is shown relative to a sample not exposed to mouse serum (Time 0 [To] positive control).

[0048] FIG. 14 shows cumulated ELISA EC50 values for the binding of antibody- peptide fusion proteins bound to various synthetic amyloid fibrils or human AL or ATTR amyloid extracts. Ec50 values were calculated using a 4-parameter logistic regression of binding curves obtained for each antibody-peptide fusion.

[0049] FIG 15A and 15B show binding of APF7, APF11, APF9, APF3, APF13, APF1, and APF5 to synthetic fibrils associated with neurodegenerative diseases, assessed by ELISA binding studies. FIG. 15A shows binding of the antibody-peptide fusion proteins to synthetic Tau fibrils. FIG. 15B shows binding of the antibody-peptide binding proteins to a-synuclein fibrils.

[0050] FIG. 16A and 16B show pHRodo red fluorescence emission levels as a measure of phagocytosis of pHRodo red-labeled rV / LWIL fibrils or human ALL amyloid extract as a percent of fluorescence measured for APF1 phagocytosis. FIG. 16A shows phagocytosis of pHRodo red-labeled rVL6WIL fibrils by antibody-peptide fusion proteins relative to antibody-peptide fusion 1 (APF1) phagocytosis of pHRodo red-labeled rVL6WIL fibrils. FIG. 16B shows phagocytosis of pHRodo red-labeled human ALL(BAL) extract relative to antibody-peptide fusion 1 (APF1) of pHRodo red-labeled human ALL(BAL) extract. FIG.9MOFO-360384346.1Attorney Docket No. 16599200164016C shows phagocytosis of pHRodo red-labeled human ATTRwt extract relative to antibody- peptide fusion 1 (APF1) of pHRodo red-labeled human ATTRwt extract.

[0051] FIG. 17 shows serum levels of APF1 and APF15 from 0 hours to 48 hours post injection (pi) in mice. APF1 and APF15 were detected using a sandwich ELISA with antihuman Fc as the capture antibody and an anti-idiotype antibody (B-13F4) as the detection reagent.

[0052] FIG. 18A and 18B show serum levels of APF1 (IgG) and bioactive APF1 (Peptide) following intravenous injection in mice (FIG. 18A) serum levels of APF1 IgG and APF1 -associated bioactive peptide (based on a heparin binding assay) following intravenous injection of APF1 into wild type mice. (FIG. 18B) serum levels of APF15 IgG and APF15- associated bioactive peptide (based on a heparin binding assay) following intravenous injection of APF15 into wild type mice.

[0053] FIG. 19A-19B show changes in serum IgG levels and bioactive peptide levels (based on heparin binding) as a percent of the first time point for APF1 (FIG. 19A) and APF15 (FIG. 19B) from 0 hours and 48 hours post injection of the antibody -peptide fusion proteins into wild type mice.

[0054] FIG. 20A and 20B show serum IgG and bioactive peptide levels (based on heparin binding) following intravenous injection, into amyloid-free non-human primates, of APF1 (FIG. 20A) and APF5 (FIG. 20B) from 0 to 24 hours post injection of the antibody - peptide fusion proteins.

[0055] FIG. 21A and 21B show percent change in serum IgG and bioactive peptide levels (based on heparin binding) levels in non-human primate serum (relative to the 1 h pi time point) over 24 h for APF1 (FIG. 21 A) and APF5 (FIG. 21B).DETAILED DESCRIPTION

[0056] Described herein is a panel of peptides capable of binding many types of amyloid. The peptides are characterized by a rigid secondary structure and a high positive charge with regularly spaced proline and basic amino acids. Using these novel amyloid-reactive peptides, peptide-protein fusions have been generated by genetically incorporating peptides into a human immunoglobulin K light chain or a human IgGl crystallizable fragment (Fc) domain. The peptides are capable of binding hypersulfated heparan sulfate and amyloid fibrils, potentially through multivalent electrostatic interactions involving the positively charged side chains. The peptides have been genetically incorporated into a human IgGlK and human10MOFO-360384346.1Attorney Docket No. 165992001640IgGlFc to develop potential therapeutic agents capable of opsonizing amyloid and facilitating clearance of the deposits. The findings provide support for the use of these novel peptides as amyloid-targeting reagents and for the peptide-immunoglobulin fusions to serve as a novel platform for the development of novel amyloid-removing therapeutics.

[0057] Provided herein are amyloid-reactive peptides that bind to human amyloid fibrils and antibody-peptide fusion proteins that bind amyloid fibrils, as well as uses thereof. The amyloid-reactive peptides are capable of binding with high affinity to human amyloid fibrils of all types and thus the antibody-peptide fusion proteins made with the amyloid-reactive peptides have pan-amyloid reactivity.

[0058] In some aspects, antibody-peptide fusion proteins comprising the novel amyloidreactive peptides of the current application display unexpected increases in in vivo stability. The increased stability of the antibody-peptide fusion proteins can contribute to increased therapeutic efficacy because they are more effective at opsonizing amyloid fibrils for a longer period of time.I. Definitions

[0059] As used herein, the singular forms “a,” “an,” and “the,” refer to both the singular as well as plural, unless the context clearly indicates otherwise. The abbreviation, “e.g.” is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation “e.g.” is synonymous with the term “for example.” As used herein, the term “comprises” means “includes.”

[0060] Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, another aspect includes from the one particular value of the range and / or to the other particular value of the range. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another aspect. In certain example embodiments, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein can be modified by the term about. Further, terms used herein such as “example,” “exemplary,” or “exemplified,” are11MOFO-360384346.1Attorney Docket No. 165992001640 not meant to show preference, but rather to explain that the aspect discussed thereafter is merely one example of the aspect presented.

[0061] It is further to be understood that all base sizes or amino acid sizes, and all molecular weight or molecular mass values, given for nucleic acids or polypeptides are approximate, and are provided for description. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. In case of conflict, the present specification, including explanations of terms, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0062] To facilitate review of the various embodiments of this disclosure, the following explanations of specific terms are provided:

[0063] The terms amyloids, amyloid deposits, amyloid fibrils, and amyloid fibers refer to insoluble fibrous protein aggregates sharing specific structural traits. The protein aggregates have a tertiary structure, for example, that is formed by aggregation of any of several different proteins and that consists of an ordered arrangement of P sheets stacked perpendicular to a fiber axis. See Sunde et al., J. Mol. Biol. (1997) 273:729-39. Abnormal accumulation of amyloids in organs may lead to amyloidosis. Although they are diverse in their occurrence, all amyloids have common morphologic properties in that they stain with specific dyes such as Congo red and have a characteristic red-green birefringent appearance in polarized light after staining. Amyloids also share common ultrastructural features and common x-ray diffraction and infrared spectra.

[0064] Amyloidosis refers to a pathological condition or disease characterized by the presence of amyloids, such as the presence of amyloid deposits. “Amyloid diseases” or “amyloidosis” are diseases associated with the formation, deposition, accumulation or persistence of amyloid fibrils. Such diseases include, but are not limited to, Alzheimer’s disease, Down’s syndrome, hereditary cerebral hemorrhage with amyloidosis of the Dutch type, and cerebral beta-amyloid angiopathy. Other amyloid diseases such as AA amyloidosis, AL amyloidosis, ATTR amyloidosis, ALect2 amyloidosis, and IAPP amyloidosis of type II diabetes are also amyloid diseases.

[0065] Systemic amyloidosis refers to forms of amyloidosis wherein amyloid accumulates in multiple organs.

[0066] Amyloidogenic refers to producing or tending to produce amyloid deposits. For example, certain soluble monomeric proteins can undergo extensive conformational changes leading to their aggregation into well-ordered, unbranching, 8- to 10-nm wide fibrils, which12MOFO-360384346.1Attorney Docket No. 165992001640 culminate in the formation of amyloid aggregates. More than thirty proteins, for example, have been found to form amyloid deposits (or amyloids) in man. Not all proteins within the class of diverse proteins, such as immunoglobulin light chains, are capable of forming amyloid, i.e., some proteins are non-amyloidogenic, meaning that they do not tend to form amyloids. Other proteins of the class, however, can form amyloid deposits and are thus amyloidogenic. Furthermore, within the class of light chain protein, some may be deemed more “amyloidogenic” than others based upon the ease with which they form amyloid fibrils. Certain light chain proteins are deemed non-amyloidogenic or less amyloidogenic because of their inability to readily form amyloid fibrils in patients or in vitro.

[0067] Clearance: The terms “clear” or “clearance” refer to reducing or removing by a measurable degree. For example, the clearance of an amyloid deposit as described herein relates to reducing or removing the deposit to a measurable or discernable degree. Clearance may result in 100% removal, but is not required to. Rather, clearance may result in less than 100% removal, such as about 10%, 20%, 30%, 40%, 50%, 60% or more removal.

[0068] Conjugate: As used herein, the term “conjugate” refers to the product of coupling or joining of two or more materials, the resulting product having at least two distinct elements, such as at least two domains. The coupled materials may be the same or may be different. Such a coupling may be via one or more linking groups. A “protein conjugate,” for example, results from the coupling of two or more amino acid sequences. A conjugate of two proteins, for example, results in a single protein that has a domain corresponding to each of the individually joined proteins.

[0069] The term “antibody” herein is used in the broadest sense and specifically covers monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments. In some aspects, an antibody comprises an Fc domain such that it is able to interact with complement and opsonize fibrils.

[0070] An “isolated” antibody is one which has been identified and separated and / or recovered from a component of its natural environment. Contaminant components of its natural environment are materials which would interfere with research, diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In some embodiments, an antibody is purified (1) to greater than 95% by weight of antibody as determined by, for example, the Lowry method, and in some embodiments, to greater than 99% by weight; (2) to a degree sufficient to obtain at least 15 residues of N- terminal or internal amino acid sequence by use of, for example, a13MOFO-360384346.1Attorney Docket No. 165992001640 spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using, for example, Coomassie blue or silver stain, Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody’s natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step.

[0071] “Native antibodies” are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains,

[0072] The term “constant region” refers to the portion of an immunoglobulin molecule having a more conserved amino acid sequence relative to the other portion of the immunoglobulin, the variable domain, which contains the antigen binding site. The constant region contains the CR1, CR2 and CR3 domains (also termed CHI, CH2, and CH3; collectively, CH) of the heavy chain and the CHL (or CL or CL1) domain of the light chain.

[0073] The “variable region” or “variable domain” of an antibody refers to the aminoterminal domains of the heavy or light chain of the antibody. The variable domain of the heavy chain may be referred to as “VH .” The variable domain of the light chain may be referred to as “VL.” These domains are generally the most variable parts of an antibody and contain the antigen-binding sites.

[0074] The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called complementarity-determining regions (CDRs) both in the light-chain and the heavychain variable domains. The more highly conserved portions of variable domains are called the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three14MOFO-360384346.1Attorney Docket No. 165992001640CDRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md, (1991)). The constant domains are not involved directly in the binding of an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.

[0075] The “light chains” of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two clearly distinct types, called kappa (“K”) and lambda (“X”), based on the amino acid sequences of their constant domains,

[0076] The term IgG “isotype” or “subclass” as used herein is meant any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions.

[0077] Depending on the amino acid sequences of the constant domains of their heavy chains, antibodies (immunoglobulins) can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 5, a, y, and p, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and described generally in, for example, Abbas et al. Cellular and Mol. Immunology, 4th ed. (W.B. Saunders, Co., 2000). An antibody may be part of a larger fusion molecule, formed by covalent or non-covalent association of the antibody with one or more other proteins or peptides.

[0078] The terms “full length antibody,” “intact antibody” and “whole antibody” are used herein interchangeably to refer to an antibody in its substantially intact form, not antibodyfragments as defined below. The terms particularly refer to an antibody with heavy chains that contain an Fc region.

[0079] A “naked antibody” for the purposes herein is an antibody that is not conjugated to a cytotoxic moiety or radiolabel.

[0080] “Antibody fragments” comprise a portion of an intact antibody, preferably comprising the antigen binding region thereof. In some embodiments, the antibody fragment described herein is an antigen-binding fragment. Examples of antibody fragments include15MOFO-360384346.1Attorney Docket No. 165992001640Fab, Fab’, F(ab’)2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0081] Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen- binding site, and a residual “Fc” fragment, whose name reflects its ability to crystallize readily. Pepsin treatment yields an F(ab’)2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen.

[0082] “Fv” is the minimum antibody fragment which contains a complete antigenbinding site. In one embodiment, a two-chain Fv species consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association, In a single-chain Fv (scFv) species, one heavy- and one light-chain variable domain can be covalently linked by a flexible peptide linker such that the light and heavy chains can associate in a “dimeric” structure analogous to that in a two-chain Fv species. It is in this configuration that the three CDRs of each variable domain interact to define an antigen- binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0083] The Fab fragment contains the heavy- and light-chain variable domains and also contains the constant domain of the light chain and the first constant domain (CHI.) of the heavy chain. Fab’ fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region. Fab’-SH is the designation herein for Fab’ in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab’)2 antibody fragments originally were produced as pairs of Fab’ fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0084] “Single-chain Fv” or “scFv” antibody fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding. For a review of scFv, see, e.g., Pluckfhun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer- Verlag, New York, 1994), pp. 269-315.

[0085] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies16MOFO-360384346.1Attorney Docket No. 165992001640 comprising the population are identical except for possible mutations, e.g., naturally occurring mutations, that may be present in minor amounts. Thus, the modifier “monoclonal.” indicates the character of the antibody as not being a mixture of discrete antibodies. In certain embodiments, such a monoclonal antibody typically includes an antibody comprising a polypeptide sequence that binds a target, wherein the target-binding polypeptide sequence was obtained by a process that includes the selection of a single target binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that a selected target binding sequence can be further altered, for example, to improve affinity for the target, to humanize the target binding sequence, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc., and that an antibody comprising the altered target binding sequence is also a monoclonal antibody of this invention. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different, determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

[0086] The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the invention may be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein, Nature, 256:495-97 (1975); Hongo et al, Hybridoma, .1.4 (3): 253-260 (.1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al, in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981 )), recombinant DNA methods (see, e.g., U.S. Pat. No. 4,8.16,567), phage-display technologies (see, e.g., Clackson et al, Nature, 352: 624-628 (1991); Marks et al, J. Mol. Biol. 222: 581-597 ( 1992); Sidhu et al, J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073- 1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467- 12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004), and technologies for producing human or human-like antibodies in animals that have parts or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits17MOFO-360384346.1Attorney Docket No. 16599200164011 et al, Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al, Nature 362: 255-258 (1993); Bruggemann et al, Year in Immunol 7:33 (1993); U.S. Pat. Nos. 5,545,807;5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al, Bio / Technology 10: 779-783 (1992); Lonberg et al, Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813 (1994) ; Fishwild et al, Nature Biotechnol. 14: 845-851 (1996); Neuberger, Nature Biotechnol 14: 826 ( 1996); and Lonberg and Huszar, Intern. Rev. Immunol 13: 65-93 (1995).

[0087] The monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (see, e.g., U.S. Pat. No. 4,816,567; and Morrison et al, Proc. Natl Acad. Sci. USA 81 :6851-6855 (1984)). Chimeric antibodies include primatized antibodies wherein the antigen-binding region of the antibody is derived from an antibody produced by, e.g., immunizing macaque monkeys with the antigen of interest.

[0088] “Humanized” forms of non-human (e.g., murine) antibodies are antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a CDR of the recipient are replaced by residues from a CDR of a non- human species (donor antibody) such as mouse, rat, rabbit, or nonhuman primate having the desired specificity, affinity, and / or capacity. In some instances, FR residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to further refine antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non- human immunoglobulin, and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see, e.g., Jones et al, Nature 321 : 522-525 (1986);Riechmann et al, Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol 2:593-596 (1992), See also, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1 : 105-11518MOFO-360384346.1Attorney Docket No. 165992001640(1998); Harris, Biochem. Soc. Transactions 23: 1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994): and U.S. Pat. Nos. 6,982,321 and 7,087,409.

[0089] A “human antibody” is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and / or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage-display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al, J. Mol. Biol., 222:581 (1991). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al, J. Immunol., 147(l):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001), Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). See also, for example, Li et al, Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology.

[0090] The term “complementarity-determining region” or “CDR,” when used herein refers to the regions of an antibody -variable domain that bind to an epitope, such as human amyloid fibrils. Generally, antibodies comprise six CDRs; three in the VH (Hl, H2, H3), and three in the VL (LI, L2, L3). In native antibodies, H3 and L3 display the most diversity of the six CDRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu n Methods in Molecular Biology 248: 1-25 (Lo, ed., Human Press, Totowa, NJ, 2003)). Indeed, naturally occurring camelid antibodies consisting of a heavy chain only are functional and stable in the absence of light chain. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993) and Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0091] A number of CDR delineations are in use and are encompassed herein. In some embodiments, the CDRs may be Kabat CDRs, which are based on sequence variability and are the most commonly used (Kabat et al., supra). In some embodiments, the CDRs may be Chothia CDRs. Chothia refers instead to the location of the structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). In some embodiments, the CDRs may be AbM CDRs. The AbM CDRs represent a compromise between the Kabat CDRs and Chothia19MOFO-360384346.1Attorney Docket No. 165992001640 structural loops, and are used by Oxford Molecular’s AbM antibody -modeling software. In some embodiments, the CDRs may be “contact” CDRs. The “contact” CDRs are based on an analysis of the available complex crystal structures. The residues from each of these CDRs are noted below.

[0092] CDRs may comprise “extended CDRs” as follows: 24-36 or 24-34 (LI), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in the VL, and 26-35 (Hl), 50-65 or 49-65 (a preferred embodiment) (H2), and 93-102, 94-102, or 95-102 (H3) in the VH. The variable-domain residues are numbered according to Kabat et al., supra, for each of these extended-CDR definitions.

[0093] “Framework” or “FR” residues are those variable domain residues other than the CDR residues as herein defined.

[0094] As use herein, the term “specifically binds to” or is “specific for” refers to measurable and reproducible interactions such as binding between a target and an antibody, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules. For example, an antibody that specifically binds to a target (which can be an epitope) is an antibody that binds this target with greater affinity, avidity, more readily, and / or with greater duration than it binds to other targets. In one embodiment, the extent of binding of an antibody to an unrelated target is less than about 10% of the binding of the antibody to the target as measured, e.g., by a radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (Kd) of < 1 M, < 100 pM, < 10 nM, < 1 nM, or < 0.1 nM. In certain embodiments, an antibody specifically binds to an epitope on a protein that is conserved among the protein from different species. In another embodiment, specific binding can include, but does not require exclusive binding.

[0095] Effective amount or Therapeutically effective amount: The amount of agent that is sufficient to prevent, treat (including prophylaxis), reduce and / or ameliorate the symptoms and / or underlying causes of any of a disorder or disease, for example to prevent, inhibit,20MOFO-360384346.1Attorney Docket No. 165992001640 and / or treat amyloidosis. In some embodiments, an “effective amount” is sufficient to reduce or eliminate a symptom of a disease. An effective amount can be administered one or more times. For example, an effective amount of a peptide is an amount that is sufficient to bind an amyloid. A peptide may be effective, for example, when parenterally administered in amounts above about 1 pg per kg of body weight to about 30 mg / kg.

[0096] Inhibit: To reduce by a measurable degree. Inhibition does not, for example, require complete loss of function or complete cessation of the aspect being measured. For example, inhibiting plaque formation can mean stopping further growth of the plaque, slowing further growth of the plaque, or reducing the size of the plaque.

[0097] Inhibiting or treating a disease: Inhibiting the full development of a disease or condition, for example, inhibiting amyloidosis. “Treatment” refers to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop. The term “ameliorating,” with reference to a disease or pathological condition, refers to any observable beneficial effect of the treatment. The beneficial effect can be evidenced, for example, by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in severity of some or all clinical symptoms of the disease, a slower progression of the disease, an improvement in the overall health or well-being of the subject, or by other parameters well known in the art that are specific to the particular disease. A “prophylactic” treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing pathology.

[0098] With regard to amyloid deposit formation, “inhibition” refers to the prevention or reduction in the formation of the amyloid deposit, such as when compared to a control. For example, inhibition may result in a reduction of about 10%, 20%, 30%, 40%, 50%, 60% or more of an amyloid deposit as compared to a control.

[0099] Label refers to any detectable compound or composition that is conjugated directly or indirectly to another molecule to facilitate detection of that molecule. Specific, non-limiting examples of labels include fluorescent tags, chemiluminescent tags, haptens, enzymatic linkages, and radioactive isotopes. A protein that is “detectably-labeled,” for example, means that the presence of the protein can be determined by a label associated with the protein.

[0100] Isolated: An “isolated” biological component, such as a peptide (for example one or more of the peptides disclosed herein), cell, nucleic acid, or serum samples has been substantially separated, produced apart from, or purified away from other biological21MOFO-360384346.1Attorney Docket No. 165992001640 components in the cell of the organism in which the component naturally occurs, for instance, other chromosomal and extrachromosomal DNA and RNA, and proteins. Nucleic acids, peptides and proteins that have been “isolated” thus include nucleic acids and proteins purified by standard purification methods. The term also embraces nucleic acids, peptides and proteins prepared by recombinant expression in a cell as well as chemically synthesized peptide and nucleic acids. The term “isolated” or “purified” does not require absolute purity; rather, it is intended as a relative term. Thus, for example, an isolated peptide preparation is one in which the peptide or protein is more enriched than the peptide or protein is in its natural environment within a cell. Preferably, a preparation is purified such that the protein or peptide represents at least 50% of the total peptide or protein content of the preparation, such as at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or even at least 99% of the peptide or protein concentration.

[0101] Join: As used herein, the term “join,” “joined,” “link,” or “linked” refers to any method known in the art for functionally connecting proteins and / or protein domains. For example, one protein domain may be linked to another protein domain via a covalent bond, such as in a recombinant fusion protein, with or without intervening sequences or domains. Joined also includes, for example, the integration of two sequences together, such as placing two nucleic acid sequences together in the same nucleic acid strand so that the sequences are expressed together.

[0102] Nucleic acid: A polymer composed of nucleotide units (ribonucleotides, deoxyribonucleotides, related naturally occurring structural variants, and synthetic non- naturally occurring analogs thereof) linked via phosphodiester bonds, related naturally occurring structural variants, and synthetic non-naturally occurring analogs thereof. Thus, the term includes nucleotide polymers in which the nucleotides and the linkages between them include non-naturally occurring synthetic analogs, such as, for example and without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide- nucleic acids (PNAs), and the like. Such polynucleotides can be synthesized, for example, using an automated DNA synthesizer. The term “oligonucleotide” typically refers to short polynucleotides, generally no greater than about 50 nucleotides. It will be understood that when a nucleotide sequence is represented by a DNA sequence (i.e., A, T, G, C), this also includes an RNA sequence (i.e., A, U, G, C) in which “U” replaces “T.”

[0103] Nucleotide includes, but is not limited to, a monomer that includes a base linked to a sugar, such as a pyrimidine, purine or synthetic analogs thereof, or a base linked to an22MOFO-360384346.1Attorney Docket No. 165992001640 amino acid, as in a peptide nucleic acid (PNA). A nucleotide is one monomer in a polynucleotide. A nucleotide sequence refers to the sequence of bases in a polynucleotide.

[0104] Conventional notation is used herein to describe nucleotide sequences: the lefthand end of a single-stranded nucleotide sequence is the 5 ‘-end; the left-hand direction of a double-stranded nucleotide sequence is referred to as the 5’-direction. The direction of 5’ to 3’ addition of nucleotides to nascent RNA transcripts is referred to as the transcription direction. The DNA strand having the same sequence as an mRNA is referred to as the “coding strand;” sequences on the DNA strand having the same sequence as an mRNA transcribed from that DNA and which are located 5’ to the 5 ’-end of the RNA transcript are referred to as “upstream sequences;” sequences on the DNA strand having the same sequence as the RNA and which are 3’ to the 3’ end of the coding RNA transcript are referred to as “downstream sequences.”

[0105] cDNA refers to a DNA that is complementary or identical to an mRNA, in either single stranded or double stranded form.

[0106] Encoding refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (for example, rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA produced by that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and non-coding strand, used as the template for transcription, of a gene or cDNA can be referred to as encoding the protein or other product of that gene or cDNA. Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. Nucleotide sequences that encode proteins and RNA may include introns.

[0107] Pharmaceutically acceptable carriers: The pharmaceutically acceptable carriers of use are conventional. Remington ’s Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, PA, 19thEdition (1995), describes compositions and formulations suitable for pharmaceutical delivery of the fusion proteins herein disclosed.

[0108] In general, the nature of the carrier will depend on the particular mode of administration being employed. For instance, parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as23MOFO-360384346.1Attorney Docket No. 165992001640 water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle. For solid compositions (e.g., powder, pill, tablet, or capsule forms), conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate. In addition to biologically neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.

[0109] Polypeptide: A polymer in which the monomers are amino acid residues that are joined together through amide bonds. When the amino acids are alpha-amino acids, either the L-optical isomer or the D-optical isomer can be used, the L-isomers being preferred. The terms “polypeptide” or “protein” as used herein is intended to encompass any amino acid sequence and include modified sequences such as glycoproteins. The term “polypeptide” is specifically intended to cover naturally occurring proteins, as well as those that are recombinantly or synthetically produced. In some examples, a peptide is one or more of the peptides disclosed herein.

[0110] Purified: The term “purified” does not require absolute purity; rather, it is intended as a relative term. Thus, for example, a purified protein preparation is one in which the protein referred to is more pure than the protein in its natural environment within a cell or within a production reaction chamber (as appropriate).[oni] Recombinant: A recombinant nucleic acid is one that has a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two otherwise separated segments of sequence. This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques.

[0112] Sequence identity: The similarity between two nucleic acid sequences, or two amino acid sequences, is expressed in terms of the similarity between the sequences, otherwise referred to as sequence identity. Sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar the two sequences are.

[0113] Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith & Waterman Adv. AppL Math. 2: 482, 1981; Needleman & Wunsch J. Mol. Biol. 48: 443, 1970; Pearson & Lipman Proc. Natl. Acad. Sci. USA 85: 2444, 1988; Higgins & Sharp Gene 73: 237-244, 1988; Higgins & Sharp CABIOS 5: 151-153, 1989; Corpet et al. Nuc. Acids Res. 16, 10881-90,24MOFO-360384346.1Attorney Docket No. 1659920016401988; Huang et al. Computer Appls. In the Biosciences 8, 155-65, 1992; and Pearson et al. Meth. Mol. Bio. 24, 307-31, 1994. Altschul etal. (J. Mol. Biol. 215:403-410, 1990), presents a detailed consideration of sequence alignment methods and homology calculations.

[0114] The NCBI Basic Local Alignment Search Tool (BLAST) (Altschul et al. J. Mol. Biol. 215:403-410, 1990) is available from several sources, including the National Center for Biotechnology Information (NCBI, Bethesda, MD) and on the Internet, for use in connection with the sequence analysis programs blastp, blastn, blastx, tblastn and tblastx.

[0115] Operably linked: A first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein-coding regions, in the same reading frame.

[0116] Pharmaceutical agent: A chemical compound or composition capable of inducing a desired therapeutic or prophylactic effect when properly administered to a subject or a cell.

[0117] Vector: A nucleic acid molecule as introduced into a host cell, thereby producing a transformed host cell. Recombinant DNA vectors are vectors having recombinant DNA. A vector can include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector can also include one or more selectable marker genes and other genetic elements known in the art. Viral vectors are recombinant DNA vectors having at least some nucleic acid sequences derived from one or more viruses. The term vector includes plasmids, linear nucleic acid molecules, and as described throughout adenovirus vectors and adenoviruses.

[0118] A subject or an individual refers to a mammal, for example, a human. The subject may be a human patient. The subject may be a mouse. A subject may be a patient suffering from or suspected of suffering from a disease or condition and may be in need of treatment or diagnosis or may be in need of monitoring for the progression of the disease or condition. The patient may also be on a treatment therapy that needs to be monitored for efficacy. In some example embodiments, a subject includes an individual suffering from, or at risk of developing an amyloid diseases, including (but not limited to), or systemic amyloidosis such as seen in patients with light chain (AL) amyloidosis, monoclonal gammopathy of unknown significance (MGUS), multiple myeloma (MM), transthyretin amyloidosis (ATTR), heart failure with preserved ejection fraction (HFpEF), heart failure with reduced ejection fraction25MOFO-360384346.1Attorney Docket No. 165992001640(HFrEF) atrial fibrillation, aortic stenosis, transthyretin variant gene carriers, and type 2 diabetes.

[0119] Preferably, residue positions which are not identical differ by conservative amino acid substitutions. The term “conservative amino acid substitutions” refer to the interchangeability of residues having similar side chains. For example, a group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains is serine and threonine; a group of amino acids having amide- containing side chains is asparagine and glutamine; a group of amino acids having aromatic side chains is phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains is lysine, arginine, and histidine; and a group of amino acids having sulfur- containing side chains is cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalaninetyrosine, lysine-arginine, alanine-valine, glutamic- aspartic, and asparagine-glutamine.

[0120] As discussed herein, minor variations in the amino acid sequences of antibodies or immunoglobulin molecules are contemplated as being encompassed by the present invention, providing that the variations in the amino acid sequence maintain at least 75%, more preferably at least 80%, 90%, 95%, and most preferably 99%. In particular, conservative amino acid replacements are contemplated. Conservative replacements are those that take place within a family of amino acids that are related in their side chains. Genetically encoded amino acids are generally divided into families: (1) acidic amino acids are aspartate, glutamate; (2) basic amino acids are lysine, arginine, histidine; (3) non-polar amino acids are alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan, and (4) uncharged polar amino acids are glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine. The hydrophilic amino acids include arginine, asparagine, aspartate, glutamine, glutamate, histidine, lysine, serine, and threonine. The hydrophobic amino acids include alanine, cysteine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, tyrosine and valine. Other families of amino acids include (i) serine and threonine, which are the aliphatic-hydroxy family; (ii) asparagine and glutamine, which are the amide containing family; (iii) alanine, valine, leucine and isoleucine, which are the aliphatic family; and (iv) phenylalanine, tryptophan, and tyrosine, which are the aromatic family. For example, it is reasonable to expect that an isolated replacement of a leucine with an isoleucine or valine, an aspartate with a glutamate, a threonine with a serine, or a similar replacement of an amino acid with a structurally related amino acid will not have a major effect on the binding or properties of the resulting molecule, especially if the replacement does not involve an amino26MOFO-360384346.1Attorney Docket No. 165992001640 acid within a framework site. Whether an amino acid change results in a functional peptide can readily be determined by assaying the specific activity of the polypeptide derivative assays are described in detail herein. Fragments or analogs of antibodies or immunoglobulin molecules can be readily prepared by those of ordinary skill in the art. Preferred amino- and carboxy -termini of fragments or analogs occur near boundaries of functional domains. Structural and functional domains can be identified by comparison of the nucleotide and / or amino acid sequence data to public or proprietary sequence databases. Preferably, computerized comparison methods are used to identify sequence motifs or predicted protein conformation domains that occur in other proteins of known structure and / or function. Methods to identify protein sequences that fold into a known three-dimensional structure are known. (Bowie et al. Science 253 : 164 (1991). Thus, the foregoing examples demonstrate that those of skill in the art can recognize sequence motifs and structural conformations that may be used to define structural and functional domains in accordance with the invention.

[0121] Preferred amino acid substitutions are those which: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for forming protein complexes, (4) alter binding affinities, and (4) confer or modify other physicochemical or functional properties of such analogs. Analogs can include various muteins of a sequence other than the naturally-occurring peptide sequence. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) may be made in the naturally- occurring sequence (preferably in the portion of the polypeptide outside the domain(s) forming intermolecular contacts). A conservative amino acid substitution should not substantially change the structural characteristics of the parent sequence (e.g., a replacement amino acid should not tend to break a helix that occurs in the parent sequence, or disrupt other types of secondary structure that characterizes the parent sequence). Examples of art-recognized polypeptide secondary and tertiary structures are described in Proteins, Structures and Molecular Principles (Creighton, Ed., W. H. Freeman and Company, New York (1984)); Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); and Thornton et al. Nature 354: 105 (1991).

[0122] With the exception of CDR1 in VH, CDRs generally comprise the amino acid residues that form the hypervariable loops. CDRs also comprise “specificity determining residues,” or “SDRs,” which are residues that contact antigen. SDRs are contained within regions of the CDRs called abbreviated-CDRs, or a-CDRs. Exemplary a-CDRs (a-CDR-Ll, a- CDR-L2, a- CDR-L3, a-CDR-Hl, a-CDR-H2, and a-CDR-H3) occur at amino acid residues27MOFO-360384346.1Attorney Docket No. 16599200164031-34 of LI, 50-55 of L2, 89-96 of L3, 31-35B of HI, 50-58 ofH2, and 95-102 of H3. (See Almagro and Fransson, Front. Biosci. 13: 1619-1633 (2008).)II. Amyloid-reactive peptides

[0123] Provided herein are amyloid-reactive peptides that bind to human amyloid fibrils. As used herein an amyloid-reactive peptide is a peptide that binds to fibrils in amyloid, including non-covalent bonding via electrostatic interactions or hydrophobic interactions. The amyloid-reactive peptides that bind to human amyloid fibrils described herein comprise periodic amino acid residues and positively charged amino acids. In some embodiments, the amyloid-reactive peptides comprise periodic proline residues and positively charged amino acids. In some embodiments, the positively charged amino acids are arginine. In some embodiments, the amyloid-reactive peptides are positively charged and bind to hypersulfated glycosaminoglycans and fibrils in amyloid. In some aspects, the periodicity of the amyloidreactive peptides facilitate the interactions between the positively charged amino acids and negatively charged fibrils and hypersulfated heparan sulfate in amyloid. In some embodiments, the amyloid-reactive peptides have a polyproline-coil structure.

[0124] As used herein amyloid-reactive is a property of a peptide that binds to human amyloid fibrils. The binding may be non-covalent bonding via electrostatic interactions or hydrophobic interactions.

[0125] As used herein, an amyloid binding motif is a periodic amino acid sequence in a amyloid- reactive peptide. The amyloid binding motif may contribute to binding of the amyloid reactive peptide to fibrils in the amyloid. The binding includes non-covalent bonding via electrostatic interactions or hydrophobic interactions.

[0126] The amyloid-reactive peptides and antibody-peptide fusion proteins comprising the amyloid-reactive peptides can be used for diagnosing and treating amyloid related disorders, such as systemic amyloidosis. Such amyloid-reactive peptides that bind to human amyloid fibrils comprise an amyloid binding motif, (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, such as any naturally occurring amino acid, wherein y is R or K, and wherein n is at least 2, and wherein the amyloid reactive-peptide binds to human amyloid fibrils with a half-maximal binding at a concentration (Ec50) that is less than about 1000 nM.

[0127] The amyloid-reactive peptides and antibody-peptide fusion proteins comprising the amyloid reactive peptides can be used for diagnosing and treating amyloid related disorders, such as systemic amyloidosis. Such amyloid-reactive peptides that bind to human amyloid fibrils comprise an amyloid binding motif, (Pxy)n (SEQ ID NO:9), wherein x is any28MOFO-360384346.1Attorney Docket No. 165992001640 amino acid, such as any naturally occurring amino acid, wherein y is R, and wherein n is at least 2, and wherein the amyloid reactive-peptide binds to human amyloid fibrils with a half- maximal binding at a concentration (Ec50) that is less than about 1000 nM.

[0128] The amyloid-reactive peptides and antibody-peptide fusion proteins comprising the amyloid reactive peptides can be used for diagnosing and treating amyloid related disorders, such as systemic amyloidosis. Such amyloid-reactive peptides that bind to human amyloid fibrils comprise an amyloid binding motif, (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, such as any naturally occurring amino acid, wherein y is K, and wherein n is at least 2, and wherein the amyloid reactive-peptide binds to human amyloid fibrils with a half- maximal binding at a concentration (Ec50) that is less than about 1000 nM.

[0129] In some embodiments, the amyloid-reactive peptides comprise an amyloid binding motif. In some embodiments, the amyloid-reactive peptides comprise an amyloid binding motif comprising a periodic proline and positively charged amino acids. In some embodiments, the positively charged amino acid is arginine.

[0130] In some embodiments, the amyloid— reactive peptide comprises an amyloid binding motif. In some embodiments, the amyloid binding motif is (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, wherein y is R or K. In some embodiments, the x is selected from a group consisting of A and G. In some embodiments, y is R. In some embodiments, the x is A. In some embodiments, the x is G. In some embodiments, the amyloid binding motif comprises PAR. In some embodiments, the amyloid binding motif comprises PGR. In some embodiments, n is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20. In some embodiments, n is between 2 and 20, 3 and 20, 4 and 20, 5 and 20, 6 and 20, 7 and 20, 8 and 20, 9 and 20, 10 and 20, 11 and 20, 12 and 20, 13 and 20, 14 and 20, 15 and 20, 16 and 20, 17 and 20, 18 and 20, or 19 and 20. In some embodiments, n is between 2 and 19, 2 and 18, 2 and 17, 2 and 16, 2 and 15, 2 and 14, 2 and 13, 2 and 12, 2 and 11, 2 and 10, 2 and 9, 2 and 8, 2 and 7, 2 and 6, 2 and 5, 2 and 4, or 2 and 3. In some embodiments, n is 9.

[0131] In some embodiments, the amyloid— reactive peptide comprises an amyloid binding motif. In some embodiments, the amyloid binding motif is (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, wherein y is R or K. In some embodiments, the x is selected from a group consisting of A and G. In some embodiments, y is K. In some embodiments, the x is G. In some embodiments, the amyloid binding motif comprises PGK. In some embodiments, n is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at29MOFO-360384346.1Attorney Docket No. 165992001640 least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20. In some embodiments, n is between 2 and 20, 3 and 20, 4 and 20, 5 and 20, 6 and 20, 7 and 20, 8 and 20, 9 and 20, 10 and 20, 11 and 20, 12 and 20, 13 and 20, 14 and 20, 15 and 20, 16 and 20, 17 and 20, 18 and 20, or 19 and 20. In some embodiments, n is between 2 and 19, 2 and 18, 2 and 17, 2 and 16, 2 and 15, 2 and 14, 2 and 13, 2 and 12, 2 and 11, 2 and 10, 2 and 9, 2 and 8, 2 and 7, 2 and 6, 2 and 5, 2 and 4, or 2 and 3. In some embodiments, n is 9.

[0132] In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif. In some embodiments, the amyloid binding motif is (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, wherein y is R or K. In some embodiments, the x is selected from a group consisting of A and G. In some embodiments, y is R. In some embodiments, the x is A. In some embodiments, the x is G. In some embodiments, the amyloid binding motif comprises PAR. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 8.

[0133] In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif. In some embodiments, the amyloid binding motif is (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, wherein y is R or K. In some embodiments, the x is selected from a group consisting of A and G. In some embodiments, y is R. In some embodiments, the amyloidreactive motif comprises PGR. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 4.

[0134] In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif. In some embodiments, the amyloid binding motif is (Pxy)n (SEQ ID NO:9), wherein x is any amino acid, wherein y is R or K. In some embodiments, the x is selected from a group consisting of A and G. In some embodiments, y is K. In some embodiments, the amyloidreactive motif comprises PGK. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 5.

[0135] In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif. In some embodiments, the amyloid binding motif is (PRx)n (SEQ ID NO: 107), wherein x is any amino acid. In some embodiments, the x is selected from a group consisting30MOFO-360384346.1Attorney Docket No. 165992001640 of A and G. In some embodiments, the amyloid-reactive motif comprises PRA. In some embodiments, the amyloid-reactive motif comprises PRG.

[0136] Also provided herein are amyloid-reactive peptides that bind to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to any of SEQ ID NOs: 1-8. In some embodiments, the amino acid substitutions are conservative substitutions. In some embodiments, the amyloid-reactive peptide comprise an amino acid selected from the groups consisting of SEQ ID NOs: 1-8. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 8.

[0137] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 1. In some embodiments, the amino acid substitutions are conservative substitutions.

[0138] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 2 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 2. In some embodiments, the amino acid substitutions are conservative substitutions.

[0139] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 3 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 3. In some embodiments, the amino acid substitutions are conservative substitutions.

[0140] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 4 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 4. In some embodiments, the amino acid substitutions are conservative substitutions.

[0141] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 5 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 5. In some embodiments, the amino acid substitutions are conservative substitutions.31MOFO-360384346.1Attorney Docket No. 165992001640

[0142] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 6 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 6. In some embodiments, the amino acid substitutions are conservative substitutions.

[0143] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 7 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 7. In some embodiments, the amino acid substitutions are conservative substitutions.

[0144] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 8. In some embodiments, the amino acid substitutions are conservative substitutions.

[0145] In some embodiments, the amyloid-reactive peptide comprises an amino acid having at least about 80%, about 85%, about 90%, about 95% or more sequence identity to any one of the amino acid sequences set forth as Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence having at least about 80%, about 85%, about 90%, about 95% or more sequence identity to any one of the amino acid sequences set forth as SEQ ID NOs: 1-8. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence having at least about 80%, about 85%, about 90%, about 95% or more sequence identity the amino acid sequences set forth as SEQ ID NOs: 4. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence having at least about 80%, about 85%, about 90%, about 95% or more sequence identity the amino acid sequences set forth as SEQ ID NOs: 8.Table 1: Amyloid reactive peptides32MOFO-360384346.1Attorney Docket No. 165992001640

[0146] In some embodiments, the amyloid-reactive peptide is a pan amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide binds to human amyloid fibrils. In some embodiments, the amyloid-reactive peptide binds to human amyloid fibrils with a Ec50 that is less than about 500 nM. In some embodiments, the amyloid-reactive peptide binds to human amyloid fibrils with a Ec50 that is from about 0.1 nM to about 500 nM. In some embodiments, the amyloid-reactive peptide binds to at least 2 types of human amyloid fibrils with a Ec50 that is less than about 500 nM. In some embodiments, the amyloidreactive peptide binds to at least 2 types of human amyloid fibrils with a Ec50 that is from about 0.1 nM to about 500 nM. In some embodiments, the amyloid-reactive peptide binds to at least 3 types of human amyloid fibrils with a Ec50 that is less than about 500 nM. In some embodiments, the amyloid-reactive peptide binds to at least 3 types of human amyloid fibrils with a Ec50 that is from about 0.1 nM to about 500 nM. In some embodiments, the amyloidreactive peptide binds to at least 4 types of human amyloid fibrils with a Ec50 that is less than about 500 nM. In some embodiments, the amyloid-reactive peptide binds to at least 4 types of human amyloid fibrils with a Ec50 that is from about 0.1 nM to about 500 nM. In some embodiments, the types of human amyloid are selected from a group consisting of rVX6WIL fibrils, ATTRwt extract, ATTRv extract, ALZ. liver extract, ALK liver extract, AP(l-40), IAAP, ALK4, AL , or ATTR fibrils. In some embodiments, the one or more human amyloid are selected from a group consisting of rVX6WIL fibrils, ATTRwt(PER125) extract, ATTRv(KEN) extract, ALX(SHI) liver extract, ALK(TAL) liver extract, AP(l-40), IAAP, ALK4, AL , ATTR, Tau or Synuclein fibrils.

[0147] In some embodiments, amyloid-reactive peptides exhibit an Ec50 binding affinity for one or more human amyloid fibrils. In some embodiments, the Ec50 binding affinity is less than about 500 nM, about 400 nM, about 300 nM, about 200 nM, about 100 nM, about 50 nM, about 40 nM, about 30 nM, about 20 nM, about 10 nM, about 5 nM, about 4 nM, about 3 nM, about 2 nM, about 1.5 nM, about 1 nM, about .5 nM, or about .1 nM. In some33MOFO-360384346.1Attorney Docket No. 165992001640 embodiments, the Ec50 binding affinity is about or less than about 100 nM. In some embodiments, the Ec50 binding affinity is about or less than 10 nM. In some embodiments, the Ec50 binding affinity is about or less than 1.5 nM. In some embodiments, the Ec50 binding affinity is from about 0.1 nM to about 500 nM. In some embodiments, the Ec50 binding affinity is from about 0.1 nM to about 500 nM, from about 0.1 nM to about 400 nM, from about 0.1 nM to about 300 nM, from about 0.1 nM to about 200 nM, or from about 0.1 nM to about 100 nM. In some embodiments, the Ec50 binding affinity is the same or different for different amyloid substrates. In some embodiments, the Ec50 binding affinity is the same or different for human amyloid substrates. In some embodiments, the Ec50 binding affinity is the same or different for synthetic amyloid substrates. In some embodiments, the one or more human amyloid are selected from a group consisting of rVL6WIL fibrils, ATTRwt extract, ATTRv extract, ALL liver extract, ALK liver extract, AP(l-40), IAAP, ALK4, ALLI , or ATTR. In some embodiments, the one or more human amyloid are selected from a group consisting of rVL6WIL fibrils, ATTRwt(PER125) extract, ATTRv(KEN) extract, ALL(SHI) liver extract, ALK(TAL) liver extract, AP(l-40), IAAP, ALK4, AL , ATTR, Tau or Synuclein fibrils.

[0148] In some embodiments, the amyloid-reactive peptide is a pan amyloid amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide binds to rVL6WIL fibrils, ATTRwt extract, ATTRv extract, ALL liver extract, ALK liver extract, AP(l-40), IAAP, ALK4, ALLI, and / or ATTR fibrils. In some embodiments, the amyloid-reactive peptide binds to rVL6WIL fibrils, ATTRwt(PER125) extract, ATTRv(KEN) extract, ALL(SHI) liver extract, ALK(TAL) liver extract, AP(l-40), IAAP, ALK4, ALLI, and / or ATTR fibrils. In some embodiments, the amyloid-reactive peptide binds to ALL, ALK, ATTRv, and / or rVL6WIL fibrils. In some embodiments, the amyloid-reactive peptide binds to Tau and / or Synuclein.

[0149] In some embodiments, the amyloid-reactive peptide binds to one or more amyloidogenic peptides in amyloids. In some embodiments, amyloids bound by the amyloidreactive peptide comprise an amyloidogenic L6 variable domain protein (VL6Wil) or an amyloidogenic immunoglobulin light chain (AL), AP(l-40) amyloid-like fibril or an amyloidogenic Ap precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloids bound by the amyloid-reactive peptide comprise amyloidogenic forms of immunoglobulin heavy chain (AH), P2-microglobulin (AP2M), transthyretin variants (ATTRv), transthyretin wild type (ATTRwt), apolipoprotein Al (AApoAI), apolipoprotein All (AApoAII), gelsolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALECT2),34MOFO-360384346.1Attorney Docket No. 165992001640 fibrinogen a variants (AFib), cystatin variants (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (Alns), prior protein (APrP); a- synuclein (AaSyn), tau (ATau), atrial natriuretic factor (AANF), ALK4, AL / other amyloidogenic peptides. The amyloidogenic peptides bound by the amyloid-reactive peptide can be a protein, a protein fragment, or a protein domain. In some embodiments, the amyloid deposits or amyloid fibrils comprise recombinant amyloidogenic proteins. In some embodiments, the amyloids are part of the pathology of a disease.

[0150] In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid deposits or human amyloid fibrils. In some embodiments, the amyloid deposits or human amyloid fibrils are located in one or more organ. In some embodiments, the amyloid deposits are located in one or more tissue type. In some embodiments, the amyloid deposits or fibrils are located in one or more of the liver, spleen, heart, kidney, brain, muscle, pancreas, stomach, upper intestine, lower intestine, and / or blood. In some embodiments, the antibody-peptide fusion proteins bind to amyloid deposits or human amyloid fibrils located in at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 organs or tissue types. In some embodiments, the antibody-peptide fusion proteins exhibit pan amyloid reactivity. In some embodiments, the antibody-peptide fusion proteins exhibit reactivity toward amyloid deposits or human amyloid fibrils located in the liver, spleen, heart, kidney, brain, muscle, pancreas, stomach, upper intestine, lower intestine, and / or blood.

[0151] In some embodiments, the amyloid-reactive peptides, comprise or consist of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69 or 70 amino acids.

[0152] In some embodiments, the amyloid-reactive peptides, comprise or consist of between about 10 and about 70 amino acids. In some embodiments, the amyloid-reactive peptides, comprise or consist of between about 10 and about 70, about 10 and about 65, about 10 and about 60, about 10 and about 55. about 10 and about 50, about 10 and about 45, about 10 and about 40, about 10 and about 35, about 10 and about 30, about 10 and about 25, about 10 and about 20, or about 10 and about 15 amino acids. In some embodiments, the amyloidreactive peptides, comprise or consist of between about 15 and about 70, about 20 and about 70, about 25 and about 70, about 30 and about 70, about 35 and about 70, about 40 and about 70, about 45 and about 70, about 50 and about 70, about 55 and about 70, about 60 and about 70, or about 65 and about 70 amino acids. In some embodiments, the amyloid-reactive35MOFO-360384346.1Attorney Docket No. 165992001640 peptides, comprise or consist of between about 20 and about 40, about 25 and about 40, about 30 and about 40, about 35 and about 40, about 20 and about 35, about 20 and about 30, or about 25 and about 30, amino acids. In some embodiments, the amyloid-reactive peptides, comprise or consist of between about 20 and about 30 amino acids.

[0153] In some embodiments, the amyloid reactive peptides have a positive charge. The positive charge contributes to binding of the amyloid reactive peptides to amyloid fibrils. In some embodiments, the positive charge is between +2 and + 20. In some embodiments, the positive charge is at least + 2, at least +3, at least +4, at least +5, at least +6, at least +7, at least +8, at least +9, at least +10, at least +11, at least +12, at least +13, at least +14, at least +15, at least +16, at least +17, at least +18, at least +19, or at least +20. In some embodiments, the positive charge is between +2 and +19, +2 and +18, +2 and +17, +2 and +16, +2 and +15, +2 and +14, +2 and +13, +2 and +12, +2 and +11, +2 and +10, +2 and +9, +2 and +8, +2 and +7, +2 and +6, +2 and +5, +2 and +4, or +2 and +3. In some embodiments, the positive charge is between +3 and +20, +4 and +20, +5 and +20, +6 and +20, +7 and +20, +8 and +20, +9 and +20, +10 and +20, +11 and +20, +12 and +20, +13 and +20, +14 and +20, +15 and +20, +16 and +20, +17 and +20, +18 and +20, or +19 and +20. In some embodiments, the positive charge is +8. In some embodiments, the positive charge is +9. In some embodiments, the positive charge is +17.

[0154] In some embodiments, the amyloid-reactive peptide has a polyproline-coil secondary structure. In some embodiments, the amyloid-reactive peptide has a spectrum reminiscent of a polyproline-coil secondary structure in PBS. In some embodiments, the amyloid-reactive peptide has a rigid secondary structure. In some embodiments, the amyloidreactive peptide is not an a-helical peptide.

[0155] The amino acids forming all or a part of the amyloid-reactive peptides may be stereoisomers and modifications of naturally occurring amino acids, non-naturally occurring amino acids, post-translationally modified amino acids, enzymatically synthesized amino acids, derivatized amino acids, constructs or structures designed to mimic amino acids, and the like. The amino acids forming the amyloid-reactive peptides of the present invention may be one or more of the 20 common amino acids found in naturally occurring proteins, or one or more of the modified and unusual amino acids. The amyloid-reactive peptide may be made by any technique known to those of skill in the art, including chemical synthesis or recombinant means using standard molecular biological techniques.III. Antibody-peptide fusion proteins36MOFO-360384346.1Attorney Docket No. 165992001640Provided herein are antibody-peptide fusion proteins that target amyloids. Such antibody - peptide fusion protein include, for example, amyloid-reactive peptides that are joined to an antibody or fragment thereof. In some embodiments, the antibody or fragment there of comprises an immunoglobulin (Ig). In some embodiments, the antibody-peptide fusion protein comprises an amyloid reactive peptide joined to the N-terminal end of the light chain, the C-terminal end of the light chain, the N-terminal end of the heavy chain and / or the C- terminal end of the heavy chain. The antibody-peptide fusion proteins can be used to treat a subject suffering from an amyloid-related disorder, such as systemic amyloidosis, for example, such as by administering the immunoglobulin-peptide fusions to the subject. In some embodiments, the antibody-peptide fusion protein is a fusion protein comprising an antibody joined to an amyloid-reactive peptide.

[0156] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, y is R. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions. In some embodiment, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1.

[0157] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, the amyloid binding motif is (PRx)n (SEQ ID NO: 107), wherein x is any amino acid. In some embodiments, x is selected from a group consisting of A and G. In some embodiments, the amyloid binding motif comprises PRA. In some embodiments, the amyloid binding motif comprises PRG.

[0158] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 1. In some embodiments, the amino acid substitutions are conservative substitutions.37MOFO-360384346.1Attorney Docket No. 165992001640

[0159] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 2 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 2. In some embodiments, the amino acid substitutions are conservative substitutions.

[0160] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 3. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 3 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 3. In some embodiments, the amino acid substitutions are conservative substitutions.

[0161] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 4 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 4. In some embodiments, the amino acid substitutions are conservative substitutions.

[0162] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 5. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 5 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 5. In some embodiments, the amino acid substitutions are conservative substitutions.

[0163] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 6. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 6 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 6. In some embodiments, the amino acid substitutions are conservative substitutions.

[0164] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino38MOFO-360384346.1Attorney Docket No. 165992001640 acid sequence set forth in SEQ ID NO: 7. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 7 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 7. In some embodiments, the amino acid substitutions are conservative substitutions.

[0165] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to SEQ ID NO: 8. In some embodiments, the amino acid substitutions are conservative substitutions.

[0166] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any non-polar amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is selected from a group consisting of A and G, wherein y is R or K, and wherein n is at least 2. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is selected from a group consisting of A and G, wherein y is R, and wherein n is at least 2. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is selected from a group consisting of A and G, wherein y is K, and wherein n is at least 2. In some embodiments, n is 9.

[0167] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is G, wherein y is R, wherein n is at least 2. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises the amino acid set forth in SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 4.

[0168] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid39MOFO-360384346.1Attorney Docket No. 165992001640 binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is G, wherein y is K, wherein n is at least 2. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises the amino acid set forth in SEQ ID NO: 5. In some embodiments, the amyloid-reactive peptide has the amino acid sequence set forth in SEQ ID NO: 5.

[0169] The antibody-peptide fusion proteins described herein comprise an amyloidreactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is A, wherein y is R, and wherein n is at least 2. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises the amino acid set forth in SEQ ID NO: 8. In some embodiments, the amyloidreactive peptide has the amino acid sequence set forth in SEQ ID NO: 8.

[0170] In some embodiments, the antibody-peptide fusion protein exhibits one or more in vivo features selected from among improved biodistribution, pan amyloid reactivity, and enhanced phagocytosis compared to a reference antibody. In some embodiments, the antibody-peptide fusion protein exhibits improved biodistribution compared to reference antibody, wherein the antibody-peptide fusion protein is detectable in organs across the body. In some embodiments, the antibody-peptide fusion protein exhibits improved biodistribution, wherein the antibody-peptide fusion protein is detectable in one or more of the liver, spleen, heart, kidney, brain, muscle, pancreas, stomach, upper intestine, lower intestine, and blood. In some embodiments, the antibody-peptide fusion protein exhibits pan amyloid reactivity compared to reference antibody, wherein the antibody-peptide fusion protein is reactive towards one or more distinct amyloid substrates in vivo. In some embodiments, the antibody- peptide fusion protein is reactive towards amyloid substrates in the liver, spleen, heart, kidney, brain, muscle, pancreas, stomach, upper intestine, lower intestine, and blood, In some embodiments, the antibody-peptide fusion protein exhibits enhanced phagocytosis compared to reference antibody, wherein contacting an amyloid substrate in vivo with the antibody- peptide fusion protein results in increased levels of phagocytosis. In some embodiments, contacting an amyloid substrate in vivo with the antibody-peptide fusion protein results in clearance of the amyloid substrate. In some embodiments, contacting an amyloid substrate in vivo with the antibody-peptide fusion protein results in enhanced phagocytosis and clearance of the amyloid substrate. In some embodiments, contacting an amyloid substrate in vivo with the antibody-peptide fusion protein provides therapeutic benefit for an individual having an amyloid related disorder. In some embodiments, the reference antibody is not engineered to bind amyloid substrates. In some embodiments, the reference antibody does not comprise an40MOFO-360384346.1Attorney Docket No. 165992001640 amyloid-reactive peptide. In some embodiments, the reference antibody is not fused to an amyloid-reactive peptide. In some embodiments, the reference antibody serves as a negative control. In some embodiments, the reference antibody is an IgG antibody. In some embodiments, the reference antibody is an IgGl, IgG2, IgG3, or IgG4 isotype. In some embodiments, the reference antibody is an IgGl isotype.

[0171] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide joined to an antibody or a fragment thereof. In some embodiments, the antibody or fragment thereof binds to human amyloid. In some embodiments, the antibody or fragment thereof binds to human immunoglobulin light chain amyloid. In some embodiments, the antibody or functional fragment thereof comprises human framework sequences. In some embodiments, the antibody or fragment thereof is humanized. In some embodiments, the antibody is a full-length antibody, a Fab fragment, or a scFv. wherein the antibody is a full-length antibody, a Fab fragment, or a scFv. In some embodiments, the antibody or fragment thereof comprises a human Fc region. In some embodiments, the human Fc region is of an IgGl, IgG2, IgG3, or IgG4 isotype.

[0172] As those skilled in the art will appreciate, the fragment antigen binding (or Fab region) is the head of an antibody that naturally interacts with target antigen. Components of the Fab region, for example, allow antibodies to bind to specific ligands and, through that interaction, to further activate the immune system. For IgG, IgA, IgD, IgE, and IgM antibody isotypes, the Ig is composed of two proteins, the heavy chain and light chain that interact in pairs to form an intact Ig comprising 2 heavy chains and 2 light chains. Both the heavy and light chains are further divided into variable domains and constant domains - the light and heavy variable domains comprising the Fab functional region and the heavy chains forming the fragment crystallizable (Fc) domains that interact with cell receptors and complement. The Fc regions of Ig bears a highly conserved N-glycosylation site.

[0173] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide; and an antibody that induces phagocytosis or acts as an opsonin. In some embodiments, an opsonin is a protein that binds to a target and induces phagocytosis of that target. In some embodiments, the opsonin comprises antibody or an antibody-peptide fusion protein. In some embodiments, the antibody-peptide fusion proteins provided herein act as opsonins by binding to amyloid and promoting phagocytosis of the amyloid. In some embodiments, the antibody comprises a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL). In some41MOFO-360384346.1Attorney Docket No. 165992001640 embodiments, the amyloid-reactive peptide and the antibody are linked at the N- and / or C- terminal end of the light chain and / or the N- and / or C-terminal end of the heavy chain. In some embodiments, the antibody-peptide fusion protein comprises two or more amyloid reactive peptides, wherein each amyloid reactive peptide is bound to the N- and / or C-terminal end of the light chain and / or the N- and / or C-terminal end of the heavy chain. In some embodiments, the antibody-peptide fusion protein comprises more than one amyloid-reactive peptide linked to the antibody. In some embodiments, the one or more amyloid-reactive peptides are linked to the antibody at the N-terminal end of one or both light chains. In some embodiments, the one or more amyloid-reactive peptides are linked to the antibody at the C- terminal end of one or both light chains. In some embodiments, the one or more amyloidreactive peptides are linked to the antibody at the N-terminal end of one or both light chain. In some embodiments, the one or more amyloid-reactive peptides are linked to the antibody at the C-terminal end of one or both heavy chains. In some embodiments, the antibody is a full length antibody. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the one or more amyloid-reactive peptides each comprises an amino acid sequence as shown in Table 1. In some embodiments, the one or more amyloid-reactive peptides each comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1.

[0174] In some embodiments, the antibody-peptide fusion protein comprises a heavy chain in N-to C-terminal direction comprising in order an amyloid reactive peptide, a spacer, a VH, a CHI, a CH2, and a CH3. In some embodiments, the spacer is any spacer or linker sequence as described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain in N- to C- terminal direction in order a VH, a CHI, a CH2, a CH3, a spacer, and an amyloid reactive peptide. In some embodiments, the antibody-peptide fusion protein comprises a light chain in N- to C- terminal direction in order an amyloid reactive peptide, a spacer, a VL, and a CL. In some embodiments, the antibody-peptide fusion protein comprises a light chain in N- to C- terminal direction in order a VL, and a CL, a spacer, and an amyloid-reactive peptide. In some embodiments, the antibody also comprises a linker amino acid sequence between the amyloid-reactive peptide and the N-terminus or C-terminus of the antibody light chain or the N- or C-terminus of the heavy chain. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 84. In some embodiments, the amyloid-reactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and the antibody-peptide fusion protein42MOFO-360384346.1Attorney Docket No. 165992001640 comprises a secretory leader sequence. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1.

[0175] In some embodiments, the antibody-peptide fusion protein comprises an antibody joined to the amyloid-reactive peptide. In some embodiments, the antibody-peptide fusion protein comprises an antibody that comprises one, two, three, four, five, or six CDRs according to SEQ ID NO: 17-22. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid-binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, the amyloidreactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any non-polar amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, n is 9. In some embodiments, y is R. In some embodiments, the amyloidreactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0176] In some embodiments, the antibody-peptide fusion protein comprises an antibody joined to the amyloid-reactive peptide. In some embodiments, the antibody-peptide fusion protein comprises an antibody that comprises one, two, three, four, five, or six CDRs according to SEQ ID NO: 17-22. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, the amyloidreactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any non-polar amino acid, wherein y is R or K, and wherein n is at least 2. In some43MOFO-360384346.1Attorney Docket No. 165992001640 embodiments, n is 9. In some embodiments, y is K. In some embodiments, the amyloidreactive peptide comprises SEQ ID NO: 5.

[0177] In some embodiments, the antibody-peptide fusion protein comprises an antibody, wherein the antibody comprises a VH that comprises (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 17, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 18, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 19, wherein the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0178] In some embodiments, the antibody-peptide fusion protein comprises an antibody, wherein the antibody comprises a VH that comprises (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 17, (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 18, and (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 19, wherein the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0179] In some embodiments, the antibody-peptide fusion protein comprises an antibody, wherein the antibody comprises a VL that comprises (a) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:20; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID44MOFO-360384346.1Attorney Docket No. 165992001640N0:21; and (c) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:22, wherein the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0180] In some embodiments, the antibody-peptide fusion protein comprises an antibody, wherein the antibody comprises a VL that comprises (a) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:20; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:21; and (c) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:22, wherein the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0181] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), wherein the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 18, and a CDR-H3 comprising45MOFO-360384346.1Attorney Docket No. 165992001640 the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0182] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), wherein the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0183] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in SEQ ID NO:20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino46MOFO-360384346.1Attorney Docket No. 165992001640 acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0184] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in SEQ ID NO:20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.47MOFO-360384346.1Attorney Docket No. 165992001640

[0185] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR- L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR- H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0186] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR- L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR- H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table48MOFO-360384346.1Attorney Docket No. 1659920016401. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0187] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:73; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, y is R. In some embodiments, x is any non-polar amino acid. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0188] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:73; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or49MOFO-360384346.1Attorney Docket No. 165992001640 antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, y is K. In some embodiments, x is any non-polar amino acid. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0189] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), the VL comprises a CDR- L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:73; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody or antibody fragment thereof is joined to an amyloid reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, the amyloid binding motif is (PRx)n (SEQ ID NO: 107), wherein x is any amino acid. In some embodiments, the x is selected from a group consisting of A and G. In some embodiments, the amyloid-reactive motif comprises PRA. In some embodiments, the amyloid binding motif comprises PRG.

[0190] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising human framework sequences. In some embodiments, the antibody is humanized. In some embodiments, the humanized antibody comprises a humanized light chain variable region (VL) as shown in Table 2. In some embodiments, the humanized antibody comprises a humanized heavy chain variable region (VH) as shown in Table 3.50MOFO-360384346.1Attorney Docket No. 165992001640Table 2: Amino Acid sequences of humanized light chain variable region sequencesTable 3: Amino Acid sequences of humanized heavy chain variable region sequences51MOFO-360384346.1Attorney Docket No. 16599200164052MOFO-360384346.1Attorney Docket No. 165992001640

[0191] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution in the FR2 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the amino acid substitution in the FR2 is selected from the group consisting of an amino acid substitution at position 36, an amino acid substitution at position 37, and an amino acid substitution at position 46, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some embodiments, the amino acid substitution in the FR2 is selected from the group consisting of Tyr at position 36, Leu at position 37, and Leu at position 46, wherein the amino acid positions are numbered according to the numbering system of Kabat.

[0192] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution in the FR3 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the amino acid substitution in the FR3 is selected from the group consisting of an amino acid substitution at position 85, and an amino acid substitution at position 87, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some embodiments, the amino acid substitution in the FR3 is selected from the group consisting of Leu at position 85, and Phe at position 87, wherein the amino acid positions are numbered according to the numbering system of Kabat.

[0193] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution in the FR2 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the amino acid substitution in the FR2 is selected from the group consisting of an amino acid substitution at position 37 and an amino acid substitution at position 48, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some embodiments, the amino acid substitution in the FR2 is53MOFO-360384346.1Attorney Docket No. 165992001640 selected from the group consisting of Vai at position 37 and Leu at position 48, wherein the amino acid positions are numbered according to the numbering system of Kabat.

[0194] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution in the FR3 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the amino acid substitution in the FR3 is selected from the group consisting of an amino acid substitution at position 67, an amino acid substitution at position 48, an amino acid substitution at position 71, an amino acid substitution at position 71, an amino acid substitution at position 76, an amino acid substitution at position 78, an amino acid substitution at position 79, an amino acid substitution at position 80, an amino acid substitution at position 89, an amino acid substitution at position 93, and an amino acid substitution at position 94, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some embodiments, the amino acid substitution in the FR3 is selected from the group consisting of Leu at position 67, Ser at position 48, Lys at position 71, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Vai at position 93, and Thr at position 94, wherein the amino acid positions are numbered according to the numbering system of Kabat.

[0195] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising one or more amino acid substitutions at one or more positions in the VL compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32, wherein the amino acid positions are numbered starting from the N-terminus of SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution at position 33 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody- peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution at position 34 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution at position 41 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized54MOFO-360384346.1Attorney Docket No. 165992001640 antibody comprises a VL comprising an amino acid substitution at position 42 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution at position 51 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution at position 90 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising an amino acid substitution at position 92 compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to a VL comprising an amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to a VL1, as shown in Table 2.

[0196] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising one or more amino acid substitutions at one or more positions in the VH compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43, wherein the amino acid positions are numbered starting from the N-terminus of SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 37 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody- peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 48 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 67 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 68 compared to a55MOFO-360384346.1Attorney Docket No. 165992001640VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 71 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 76 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 78 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 79 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 80 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 92 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 96 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising an amino acid substitution at position 97 compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid substitutions compared to a VH comprising an amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid substitutions compared to VH1, as shown in Table 3.

[0197] In some embodiments, the antibody-peptide fusion comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising one or more amino56MOFO-360384346.1Attorney Docket No. 165992001640 acid residues at one or more positions in the VL, wherein the amino acid positions are numbered starting from the N-terminus of the VL. In some embodiments, the antibody- peptide fusion comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Ser, Gin, Glu, His, or Ala at position 33, Ala or Vai at position 34, Tyr at position 41, Leu at position 42, Leu at position, 51 Leu at position 90 and / or Phe at position 92.

[0198] In some embodiments, the antibody-peptide fusion comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising one or more amino acid residues at one or more positions in the VH, wherein the amino acid positions are numbered starting from the N-terminus of the VH. In some embodiments, the antibody- peptide fusion comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising Vai at position 37, a Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, Thr at position 92, Vai at position 96, and / or Thr position 97. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising one or more amino acid substitutions at one or more positions in the VL, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and / or Phe at position 87.

[0199] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising one or more amino acid residues at one or more positions in the VH, wherein the amino acid positions are numbered according to the numbering system of Kabat. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH comprising Vai at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Vai at position 93, and / or Thr at position 94.

[0200] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and a VH57MOFO-360384346.1Attorney Docket No. 165992001640 comprising Vai at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Vai at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Leu at position 46 and Phe at position 87, and a VH comprising Leu at position 48, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the antibody- peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Leu at position 46 and Phe at position 87, and a VH comprising Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Vai at position 78, Leu at position 79, Vai at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Leu at position 46 and Phe at position 87, and a VH comprising Lys at position 71, Ser at position 76, Vai at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VL comprising Leu at position 46 and Phe at position 87, and a VH comprising Lys at position 71, Vai at position 78, Leu at position 79, Vai at position 93, and Thr at position 94.

[0201] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32- 42. In some embodiments, the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising the VL of VL4 as shown in Table 2, and the VH of VH9 as shown in Table 3. In some embodiments, the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, y is R. In some embodiments, x is any nonpolar amino acid. In some embodiments, n is 9. In some embodiments, the amyloid-reactive58MOFO-360384346.1Attorney Docket No. 165992001640 peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0202] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32- 42. In some embodiments, the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising the VL of VL4 as shown in Table 2, and the VH of VH9 as shown in Table 3. In some embodiments, the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, y is K. In some embodiments, x is any nonpolar amino acid. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0203] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO:55. In some embodiments, the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any nonpolar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some59MOFO-360384346.1Attorney Docket No. 165992001640 embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0204] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO:55. In some embodiments, the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any nonpolar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0205] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4.

[0206] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0207] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:34 and the VH comprises the60MOFO-360384346.1Attorney Docket No. 165992001640 amino acid sequence set forth in SEQ ID NO:48. In some embodiments, the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any nonpolar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0208] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:34 and the VH comprises the amino acid sequence set forth in SEQ ID NO:48. In some embodiments, the antibody is joined to an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any nonpolar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0209] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:34 and the VH comprises the amino acid sequence set forth in SEQ ID NO:48. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4.

[0210] In some embodiments, the antibody-peptide fusion protein comprises an antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL)61MOFO-360384346.1Attorney Docket No. 165992001640 and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:34 and the VH comprises the amino acid sequence set forth in SEQ ID NO:48. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0211] In certain embodiments, the antibody-peptide fusion protein may include linker and / or spacer sequences of amino acids between the C- or N-terminus of the light chain or C- or N-terminus of the heavy and the amyloid-reactive peptide. In certain embodiments, the antibody-peptide fusion proteins may include linker and / or spacer sequences of amino acids between the N-terminal of the amyloid-reactive peptide and a leader sequence required for secretion of antibody-peptide fusion protein from cells expressing the reagent.

[0212] In some embodiments, the linker and / or spacer is uncharged. In some embodiments a linker and / or spacer peptide may comprise or consist of from about 3 to about 55 amino acids. The linker and / or spacer peptides of the present invention may comprise or consist of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 amino acids. As used herein, a nucleic acid sequence or amino acid sequence is “adjacent” to another nucleic acid sequence or amino acid sequence if such nucleic acid sequences or amino acid sequences are close to each other in sequence. For example, two nucleic acid sequences can be adjacent to each other as described herein but still include an intervening linker and / or spacer sequence.

[0213] In some embodiments, the amyloid-reactive peptide and antibody or fragment thereof are joined via a linker. In some embodiments, the linker may comprise an amino-acid set forth in SEQ ID NOs: 82. In some embodiments, the amyloid-reactive peptide and antibody or fragment thereof comprise an N-terminal spacer. In some embodiments, the spacer is an N-terminal spacer. In some embodiments, the N-terminal spacer comprises an amino acid sequence set forth in SEQ ID: NO: 83 or SEQ ID NO: 103.

[0214] In some embodiments, the amyloid-reactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and the antibody-peptide fusion protein comprises a secretory leader sequence. In some embodiments, the secretory leader sequence comprises an amino acid sequence set forth in SEQ ID NO: 84.

[0215] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the62MOFO-360384346.1Attorney Docket No. 165992001640 light chain. In some embodiments, the amyloid-reactive peptide is linked to the C-terminus of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the light chain with the amyloid-reactive peptide fused to the C-terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 86, 87, and 93-97.

[0216] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the light chain with the amyloid-reactive peptide fused to the C-terminus of the light chain comprises an C-terminal tyrosine residue. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the light chain with the amyloid-reactive peptide fused to the C-terminus of the light chain comprising the C-terminal tyrosine residue comprises an amino acid sequence selected from any one of SEQ ID NOs: 93, 95, or 97.

[0217] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the light chain with the amyloid-reactive peptide63MOFO-360384346.1Attorney Docket No. 165992001640 fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NO: 93 or 86.

[0218] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO:55.

[0219] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 86.

[0220] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 93.64MOFO-360384346.1Attorney Docket No. 165992001640

[0221] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 87.

[0222] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain comprises the amino acid sequence set forth in SEQ ID NO: 97.

[0223] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain with the amyloid-reactive peptide fused to the C-terminus of the light chain comprises the amino acid sequence set forth in SEQ ID NO:95.

[0224] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain with the amyloid-reactive peptide65MOFO-360384346.1Attorney Docket No. 165992001640 fused to the C-terminus of the light chain comprises the amino acid sequence set forth in SEQ ID NO:96.

[0225] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody-peptide fusion protein comprises a secretory leader sequence. In some embodiments, the secretory leader sequence comprises the amino acid sequence test forth in SEQ ID NO: 84. In some embodiments, the secretory leader sequence is joined to the amyloid-reactive peptide via a N-terminal spacer. In some embodiments, the N-terminal spacer comprises the amino acid sequence set forth in SEQ ID NO: 83. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the light chain with the amyloid-reactive peptide fused to the N- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 85, 89, 91, and 92.

[0226] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the light chain with the amyloid-reactive peptide fused to the N- terminus of amyloid-reactive peptide comprises a tyrosine reside at the N-terminus of the amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody-peptide fusion protein comprises a secretory leader sequence. In some embodiments, the secretory leader sequence comprises the amino acid sequence test forth in SEQ ID NO: 84. In some embodiments, the secretory leader sequence is joined to the amyloid-reactive peptide via a N- terminal spacer. In some embodiments, the N-terminal spacer comprises the amino acid sequence set forth in SEQ ID NO: 83. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared66MOFO-360384346.1Attorney Docket No. 165992001640 to the amino acid sequences shown in Table 1. In some embodiments, the light chain with the amyloid-reactive peptide fused to the N-terminus of the light chain comprising the N-terminal tyrosine residue comprises an amino acid sequence selected from any one of SEQ ID NOs: 89 or 91.

[0227] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH). In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:55 and the VH comprises the amino acid sequence set forth in SEQ ID NO:36.

[0228] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain with the amyloid-reactive peptide fused to the N-terminus of the light chain comprises the amino acid sequence set forth in SEQ ID NO:85.

[0229] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain with the amyloid-reactive peptide67MOFO-360384346.1Attorney Docket No. 165992001640 fused to the N-terminus of the light chain comprises the amino acid sequence set forth in SEQ ID NO:89.

[0230] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain with the amyloid-reactive peptide fused to the N-terminus of the light chain comprises the amino acid sequence set forth in SEQ ID NO:91.

[0231] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the N-terminal end of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a linker. In some embodiments, the linker comprises the amino acid sequence set forth in SEQ ID NO: 82. In some embodiments, the antibody or fragment thereof comprising a light chain and a heavy chain. In some embodiments, the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 104 and the light chain with the amyloid-reactive peptide fused to the N-terminus of the light chain comprises the amino acid sequence set forth in SEQ ID NO:92.

[0232] In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and antibody or fragment thereof joined together at the C-terminal end of the heavy chain. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the heavy chain with the amyloid-reactive peptide fused to the C-terminus of the heavy chain comprises an amino acid sequence set forth in SEQ ID NO: 88.

[0233] In some embodiments, the antibody-peptide fusion proteins described herein are more stable than antibody-peptide fusion proteins previously described. In some embodiments, stability of the antibody-peptide fusion proteins refers to in vivo stability of the68MOFO-360384346.1Attorney Docket No. 165992001640 amyloid-reactive peptide and / or antibody or fragment thereof. In some embodiments, in vivo stability is measured using a pharmacokinetics (PK) profile. Without being bound to this theory, the increased stability of the antibody-peptide fusion proteins described herein may be based on the primary structure (amino acid sequence) of the amyloid-reactive peptides. The inclusion of the proline amino acids may hinder the activity of proteases that can degrade the amyloid-reactive peptides.

[0234] In some embodiments, the amyloid-reactive peptides are less likely to be cleaved from the antibody or fragment thereof when the antibody-peptide fusion protein is administered to a subject. In some embodiments, the lower cleavage rate may contribute to an increased in vivo stability compared antibody-peptide fusion proteins previously described. In some embodiments, the antibody-peptide fusion proteins are stable in vivo for at least 24 hours.

[0235] In some embodiments, about 24 hours after injection of the antibody-peptide fusion protein to a subject, about 45% of the antibody-peptide fusion protein is bioactive in blood collected from a subject. In some embodiments, about 48 hours after injection of the antibody-peptide fusion protein to a subject, about 11% of the antibody-peptide fusion protein is bioactive in blood collected from a subject. Without being bound to this theory, the amount of the antibody-peptide fusion protein remaining after injection is due to the PK of the antibody-peptide fusion. In some embodiments, the PK is due to the natural clearance of the antibody. In some embodiments, the antibody-peptide fusion protein detectable in blood is characterized by bioactivity of the antibody-bound amyloid-reactive peptide relative to the antibody or fragment thereof forming the antibody-peptide fusion protein.

[0236] In some embodiments, about 1 hours after injection of the antibody-peptide fusion protein to a subject, about 100% of the antibody-peptide fusion protein is bioactive in blood collected from a subject. In some embodiments, about 2 hours after injection of the antibody- peptide fusion protein to a subject, about 80% of the antibody-peptide fusion protein is bioactive in blood collected from a subject. In some embodiments, about 8 hours after injection of the antibody-peptide fusion protein to a subject, about 50% of the antibody- peptide fusion protein is bioactive in a blood sample collected from a subject. In some embodiments, about 24 hours after injection of the antibody-peptide fusion protein to a subject, about 10% of the antibody-peptide fusion protein is bioactive in blood collected from a subject. In some embodiments, the antibody-peptide fusion protein detectable in blood is69MOFO-360384346.1Attorney Docket No. 165992001640 characterized by bioactivity of the amyloid-reactive peptide and the antibody or fragment thereof from the antibody-peptide fusion protein.

[0237] In some embodiments, a difference in PK for the antibody -bound amyloidreactive peptides and the antibody or fragment thereof that make up the antibody-peptide fusion proteins is not detected. In some embodiments, the antibody-bound amyloid-reactive peptide and antibody or fragment thereof are detectable for up to about 24 or about 48 hours post injection (pi). In some embodiments, similar concentrations of the antibody -bound amyloid-reactive peptides and the antibody or fragment thereof that make up the antibody- peptide fusion protein are detected for up to about 24 or about 48 hours PI.

[0238] Also provided herein are pharmaceutical compositions comprising any of the amyloid-reactive peptide and / or antibody-peptide fusion proteins described herein. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions include at least one additional therapeutic agent.

[0239] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier may be a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, or a combination thereof.

[0240] In various embodiments, the pharmaceutical compositions according to the disclosure may be formulated for delivery via any route of administration. This may include e.g., aerosol, nasal, oral, transmucosal, transdermal, parenteral or enteral. In some embodiments, administration is intravenous or subcutaneous.

[0241] Provided are pharmaceutical formulations including the amyloid-reactive peptide and / or antibody-peptide fusion proteins described herein. The pharmaceutical compositions and formulations generally include one or more optional pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical compositions include at least one additional therapeutic agent.

[0242] The term “pharmaceutical formulation” refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered.70MOFO-360384346.1Attorney Docket No. 165992001640

[0243] Buffering agents in some aspects are included in the compositions. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more buffering agents is used. The buffering agent or mixtures thereof are typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0244] In certain example embodiments, the amyloid-reactive peptides and / or antibody- peptide fusion protein may be obtained by isolation or purification. Protein purification techniques involve, at one level, the homogenization and crude fractionation of cells, tissue, or organ to peptide and non-peptide fractions. Other protein purification techniques include, for example, precipitation with ammonium sulfate, polyethylene glycol (PEG), antibodies and the like, or by heat denaturation, followed by: centrifugation; chromatography steps such as ion exchange, gel filtration, reverse phase, hydroxylapatite and affinity chromatography; isoelectric focusing; gel electrophoresis, for example polyacrylamide gel electrophoresis; and combinations of these and other techniques.

[0245] Various chromatographic techniques include but are not limited to ion-exchange chromatography, gel exclusion chromatography, affinity chromatography, immuno-affinity chromatography, and reverse phase chromatography. A particularly efficient method of purifying peptides is fast performance liquid chromatography (FPLC) or even high- performance liquid chromatography (HPLC).IV. Diagnostic and detection methods

[0246] Also provided herein are methods of identifying an amyloid deposit in a subject. In some embodiments, the methods are for diagnosing an amyloid-related disorder in the subject based on the presence of an amyloid deposit.

[0247] In certain example embodiments, the antibody-peptide fusion proteins or amyloidreactive peptides can be labeled with various agents to allow their detection in vivo and in in vitro assays, such as after the fusion peptides are purified. Without being limited this may include radionuclides (e.g., I-125, 1-123, 1-124, 1-131, Zr-89, Tc-"m, Cu-64, Br-76, F-18); enzymes (horse radish peroxidase); biotin; fluorophores, etc. Any means known in the art for detectably labeling a protein can be used and / or adapted for use with the methods described71MOFO-360384346.1Attorney Docket No. 165992001640 herein. For example, the antibodies or fragments thereof, and / or the amyloid-reactive peptides, can be radiolabeled with a radioisotope, or labeled with a fluorescent tag or a chemiluminescent tag. Example radioisotopes include, for example,18F,U 1ln, "mTc, Zr89, I124,123I, and125I. These and other radioisotopes can be attached to the isolated immunoglobulin light or heavy chain using well known chemistry that may or not involve the use of a chelating agent, such as DTPA or DOTA covalently linked to the light or heavy chain protein of the antibody, for example. Example fluorescent or chemiluminescent tags include fluorescein, Texas red, rhodamine, Alexa dyes, and luciferase that can be conjugated to the protein by reaction with lysine, cysteine, glutamic acid, and aspartic acid side chains. In one example embodiment, the label is detected using a fluorescent microplate reader, or fluorimeter, using the excitation and emission wavelengths appropriate for the tag that is used. Radioactive labels can be detected, for example, using a gamma or scintillation counter depending on the type of radioactive emission and by using energy windows suitable for the accurate detection of the specific radionuclide. However, any other suitable technique for detection of radioisotopes can also be used to detect the label. In some embodiments, the detection is in vitro. In some embodiments, the detection is in vivo.

[0248] With regard to amyloidosis, such labeling, for example, can be used to diagnose the presence of amyloid, to determine the amyloid protein load, to monitor the ability of the modified immunoglobulin, humanized antibodies, or antibody-peptide fusion proteins to bind amyloid in a particular subject, to monitor the progression of amyloidosis, and / or to monitor a subject’s response to an amyloid treatment (including treatments associated with the administration of the antibody-peptide fusion proteins to the subject). For example, antibody- peptide fusion protein and / or amyloid-reactive peptide are labeled with a detectable label as described herein and thereafter administered to a subject that is suffering from, or suspected to be suffering from, an amyloid-related disorder disease (e.g., amyloidosis, monoclonal gammopathy of unknown significance (MGUS), multiple myeloma (MM), or related plasma cell diseases). Thereafter, the subject can be imaged, for example, to detect the presence of the detectably-labeled antibody-peptide fusion proteins and / or detectably-labeled amyloidreactive peptides.

[0249] In certain example embodiments, the signals from the detectably-labeled amyloidreactive peptide can be quantified, thereby providing an indication of the level of amyloid deposit in the subject. For example, the signal intensity may be compared to a standard signal threshold, above which amyloidosis is present but below which amyloidosis is absent or at a72MOFO-360384346.1Attorney Docket No. 165992001640 low level. The subject can be diagnosed as having amyloid, in which case a treatment can be administered, such as such as chemotherapy, corticosteroid medicines (lenalidomide or thalidomide) and / or bortezomib (Velcade). Additionally or alternatively, the amyloid-reactive peptide described herein can be administered to the subject in an effort to treat the subject as described herein. In certain example embodiments, the subject may be stratified into one or more groups, such as a low amyloid load, medium amyloid load, or high amyloid load, and then treated accordingly. To monitor treatment progress, the subject may be re-administered the detectably-labeled amyloid-reactive peptide, and hence reassessed for their amyloid load.

[0250] In some embodiments, the amyloid-related disorder is systemic amyloidosis or localized amyloidosis. In some embodiments, the amyloid-related disorder is selected from the group including, but not limited to, AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, APrP, and Ap amyloidosis. In some embodiments, the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

[0251] In some embodiments, the amyloid-related disorder is systemic amyloidosis. In some embodiments, the amyloid-related disorder is selected is selected from the group including, but not limited to, AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, and ACys, In some embodiments, the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

[0252] In some embodiments, the amyloid reactive peptide or antibody-peptide fusion protein is conjugated to a detectable label. In some embodiments, the detectable label is selected from the group consisting ofnC,18F,inIn, "mTc,89Ga,123I,124I,1251,89Zr, a biotin or a fluorophore. In some embodiments, the detectable label is124I. In some embodiments, the detectable label is "mTc.

[0253] In some embodiments, the antibody-peptide fusion protein conjugated to a detectable label comprises an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide is conjugated to a detectable label. In some embodiments, the amyloidreactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, n is 9. In some embodiments, the amyloidreactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive73MOFO-360384346.1Attorney Docket No. 165992001640 peptide comprises SEQ ID NO: 8. In some embodiments, the detectable label is124I. In some embodiments, the detectable label is "mTc.

[0254] In some embodiments, the methods comprise identifying an amyloid deposit in a subject, comprising administering an amyloid-reactive peptide conjugated to a detectable label or an amyloid-reactive peptide conjugated to a detectable label and detecting a signal from the detectable label. In some embodiments, the subject is diagnosed with an amyloid related disorder if a signal is detected. In some embodiments, a subject is treated for the amyloid-related disorder if the signal is detected.

[0255] In some embodiments, the amyloid-reactive peptides are conjugated to a detectable label. In some embodiments, the detectable label is conjugated to the C-terminus and / or N-terminus of the amyloid-reactive peptide. In some embodiments, the detectable label is selected from the group consisting ofnC,18F,niIn, "mTc,89Ga,123I,124I,1251,89Zr, a biotin or a fluorophore. In some embodiments, the detectable label is124I. In some embodiments, the detectable label is "mTc. In some embodiments, the amyloid reactive- peptide comprises a biotinylated N-terminal.

[0256] In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises SEQ ID NO: 8. In some embodiments, the detectable label is124I. In some embodiments, the detectable label is "mTc.

[0257] In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide conjugated to a detectable label comprises SEQ ID NO: 5. In some embodiments, the detectable label is124I. In some embodiments, the detectable label is "mTc.74MOFO-360384346.1Attorney Docket No. 165992001640

[0258] In some embodiments, the subject is a mammal such as primate, bovine, rodent, or pig. In some embodiments, the subject is a human.V. Methods of treatment

[0259] Also provided herein are methods of treating a subject having an amyloid related disorder, comprising administering to the subject an effective amount of an antibody-peptide fusion protein of the present disclosure.

[0260] In some embodiments, an effective amount of antibody-peptide fusion protein described herein is administered to a subject, thereby treating the subject. In certain example aspects, provided is a method for clearing amyloid deposits in a subject. The method includes, for example, selecting a subject with amyloidosis and administering to the subject an effective amount of an antibody-peptide fusion protein as described herein. The antibody- peptide fusion protein include, for example, an amyloid-reactive peptide that binds to amyloid deposits that is joined with an antibody or fragment thereof through the N-terminal of the light chain protein or the C-terminal of the heavy chain of the antibody or fragment thereof. Administration of the antibody-peptide fusion peptide thereby results in clearance of the amyloid and hence treatment of the subject.

[0261] In some embodiments, the antibody-peptide fusion proteins bind amyloids in an individual. In some embodiments, the amyloid deposits may contribute to the pathology of a disease. In other embodiments, the amyloid deposits may be indicative of amyloidosis or an amyloid-related disorder in an individual. In some embodiments, the antibody-peptide fusion proteins bind to amyloids in an individual with an amyloidosis. In some embodiments, the amyloidosis is localized to a specific tissue or organ system, such as the liver, the heart, or the central nervous system. In other embodiments, the amyloidosis is a systemic amyloidosis. In some embodiments, the amyloidosis is a familial amyloidosis. In other embodiments, the amyloidosis is a sporadic amyloidosis. In some embodiments, the amyloidosis or amyloid- related disease selected from the group including, but not limited to, AA amyloidosis, AL amyloidosis, AH amyloidosis, Ap amyloidosis, ATTR amyloidosis, ALECT2 amyloidosis, and IAPP amyloidosis of type II diabetes, , ATTRwt, familial hemodialysis, senile systemic aging, aging pituitary disorder, iatrogenic syndrome. In some embodiments, the antibody- peptide fusion proteins bind to amyloids associated with normal aging. In other embodiments, the antibody-peptide fusion proteins are used in the diagnosis, treatment, or prognosis of an amyloidosis or amyloid-related disorder in a subject.75MOFO-360384346.1Attorney Docket No. 165992001640

[0262] In some embodiments, the amyloid-related disorder is systemic amyloidosis or localized amyloidosis. In some embodiments, the amyloid-related disorder is selected from including, but not limited to AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, APrP, , and Ap amyloidosis. In some embodiments, the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

[0263] In some embodiments, the amyloid-related disorder is systemic amyloidosis. In some embodiments, the amyloid-related disorder is selected is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, and ACys, In some embodiments, the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

[0264] In some embodiments, the methods comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0265] In some embodiments, the methods comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ76MOFO-360384346.1Attorney Docket No. 165992001640ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 5.

[0266] In some embodiments, the methods comprise treating AL amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody -peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0267] In some embodiments, the methods comprise treating AL amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0268] In some embodiments, the methods comprise treating ATTR amyloidosis. In some embodiments, the methods comprise administering an affecting amount of an antibody- peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide. In some embodiments, the amyloid-reactive77MOFO-360384346.1Attorney Docket No. 165992001640 peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments the amyloidreactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0269] In some embodiments, the methods comprise treating ATTR amyloidosis. In some embodiments, the methods comprise administering an affecting amount of an antibody- peptide fusion protein to the subject. In some embodiments, the antibody -peptide fusion protein comprises an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments the amyloidreactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0270] In some embodiments, the methods comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein is APF4.

[0271] In some embodiments, the methods comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein is APF5.

[0272] In some embodiments, the methods comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the methods78MOFO-360384346.1Attorney Docket No. 165992001640 comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein is APF11.

[0273] In some embodiments, the methods comprise treating systemic amyloidosis in a subject. In some embodiments, the methods comprise administering an affecting amount of an antibody-peptide fusion protein to the subject. In some embodiments, the antibody-peptide fusion protein is APF12.

[0274] In some embodiments, the antibody-peptide fusion protein is administered intravenous or subcutaneous administration.

[0275] In some embodiments, the antibody-peptide fusion protein is administered in sufficient amounts to induce phagocytosis of the amyloid by cells of the immune system (e.g., macrophages).

[0276] In some embodiments, treating an amyloid related disorder comprises clearing an amyloid deposit. In some embodiments, the amyloid deposit is removed. In some embodiments, the amyloid deposit is opsonized by the antibody-peptide fusion protein. In some embodiments, binding of the antibody-peptide fusion protein to human amyloid fibrils promotes the phagocytosis of the human amyloid fibrils and the removal of the amyloid deposit. In some embodiments, the antibody-peptide fusion protein opsonizes human amyloid fibrils, thereby removing of the amyloid deposit. In some embodiments, binding of the antibody-peptide fusion protein to human amyloid fibrils promotes the phagocytosis and / or opsonization of human amyloid fibrils to an equal or greater extent than a control antibody.

[0277] In some embodiments, the subject is a mammal such as primate, bovine, rodent, or pig. In some embodiments, the subject is a human.VI. Nucleic acids, vectors, host cells, and methods of making peptide-antibody fusion proteins and amyloid-reactive peptides

[0278] Also provided herein are nucleic acid(s) encoding an antibody-peptide fusion protein and / or amyloid reactive peptide of the present disclosure. The antibody-peptide fusion protein may be any of the antibody-peptide fusion proteins described herein. The amyloidreactive peptide may be any of the amyloid-reactive peptides described herein.

[0279] In some embodiments, the nucleic acid provided herein are in one or more vectors. For example, in some embodiments, provided herein is a vector comprising a heavy79MOFO-360384346.1Attorney Docket No. 165992001640 chain and light chain of an antibody, wherein the light chain is linked to a peptide. In some embodiments, the heavy chain and the light chain linked to a peptide are in different vectors.

[0280] In some embodiments, the vector comprises the nucleic acid(s) encoding an antibody-peptide fusion protein of the present disclosure. In some embodiments, the vector comprises the nucleic acid(s) encoding an amyloid-reactive peptide of the present disclosure.

[0281] For antibody production, the heavy chain and light chain linked to a peptide expression vectors may be introduced into appropriate production cell lines know in the art. Introduction of the expression vectors may be accomplished by co-transfection via electroporation or any other suitable transformation technology available in the art. Antibody producing cell lines can then be selected and expanded and antibodies purified. The purified antibodies can then be analyzed by standard techniques such as SDS-PAGE.

[0282] Also provided is a host cell comprising a nucleic acid encoding any of the antibody-peptide fusion proteins and / or amyloid-reactive peptides described herein. Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells described herein. For example, the antibody-peptide fusion protein may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed. For expression of antibody fragments and polypeptides in bacteria, see, e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, describing expression of antibody fragments in E. coh.) After expression, the antibody linked to a peptide may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.

[0283] In some embodiments, the host cell comprising a vector comprising a nucleic acid(s) encoding an antibody-peptide fusion protein of the present disclosure. In some embodiments, the host cell comprising a vector comprising a nucleic acid(s) encoding an amyloid-reactive peptides of the present disclosure.

[0284] Suitable host cells for the expression of glycosylated antibody are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

[0285] Plant cell cultures can also be utilized as hosts. See, e.g., US Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIES™ technology for producing antibodies in transgenic plants).80MOFO-360384346.1Attorney Docket No. 165992001640

[0286] Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et cd., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse Sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3 A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, c.g, in Mather et al., Annals N Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0 and Sp2 / 0. For a review of certain mammalian host cell lines suitable for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).VII. Purification Methods

[0287] Also provided herein are methods of making an antibody-peptide fusion protein of the present disclosure. In some embodiments, the method comprises culturing a host cell of the present disclosure under conditions suitable for expression of the vector encoding the antibody-peptide fusion protein and recovering the antibody-peptide fusion protein.

[0288] In some embodiments, the method of making an antibody-peptide fusion protein comprises i) culturing a host cell comprising a vector encoding an antibody-peptide fusion protein under perfusion cell culture conditions suitable for expression of the antibody-peptide fusion protein; and ii) recovering the antibody-peptide fusion protein about every 12-36 hours. In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and an antibody or fragment thereof. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.81MOFO-360384346.1Attorney Docket No. 165992001640

[0289] In some embodiments, the method of making an antibody-peptide fusion protein comprises i) culturing a host cell comprising a vector encoding an antibody-peptide fusion protein under perfusion cell culture conditions suitable for expression of the antibody-peptide fusion protein; and ii) recovering the antibody-peptide fusion protein about every 12-36 hours. In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and an antibody or fragment thereof. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0290] In some embodiments, the method comprises culturing a host cell using a fed- batch culture method. Fed batch culture refers to a method of culturing cells, wherein the cell culture is supplemented with fresh medium, i.e., the cells are “fed” with new medium while spent medium is not removed. Typically, “fed-batch” culture process is performed in a bioreactor and additional components (e.g., nutritional supplements) are added to the culture at some time after initiation of the culture process. The controlled addition of nutrients directly affects the growth rate of the culture and allows for avoidance of the buildup of overflow metabolites (see, for example, Wlaschin, K. F. et al., “Fedbatch culture and dynamic nutrient feeding,” Cell Culture Engineering, 101 :43-74 (2006) and Lee, J. et al., “Control of fed-batch fermentations,” Biotechnol. Adv., 17:29-48 (1999)). A fed-batch culture is typically terminated at some point and the cells and / or components in the medium are harvested and optionally purified.

[0291] In some embodiments, the method comprises culturing a host cell using a perfusion culture method. Perfusion refers to a method of culturing cells, wherein additional fresh medium is provided to the culture and spent medium is removed from the culture. Perfusion is initiated after the culture is seeded and can occur either continuously or intermittently, as desired, over a period of time. The fresh medium added during perfusion typically provides nutritional supplements for the cells that have been depleted during the culturing process. Perfusion also allows for removal of cellular waste products and toxic byproducts from the cell culture. Perfusion is performed during the growth phase of the cells, but can also be continued after the cells have been transferred to a fed-batch cell culture.82MOFO-360384346.1Attorney Docket No. 165992001640

[0292] In some embodiments, the method of making an antibody-peptide fusion protein comprises culturing a host cell comprising a vector encoding an antibody-peptide fusion protein under perfusion cell culture conditions suitable for expression of the antibody-peptide fusion protein. In some embodiments, the method comprises using a fed-batch culture method for the production of the antibody-peptide fusion protein. In some embodiments, the method comprises using a perfusion culture method for the production of the antibody-peptide fusion protein. In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and an antibody or fragment thereof. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0293] In some embodiments, the method of making an antibody-peptide fusion protein comprises culturing a host cell comprising a vector encoding an antibody-peptide fusion protein under perfusion cell culture conditions suitable for expression of the antibody-peptide fusion protein. In some embodiments, the method comprises using a fed-batch culture method for the production of the antibody-peptide fusion protein. In some embodiments, the method comprises using a perfusion culture method for the production of the antibody-peptide fusion protein. In some embodiments, the antibody-peptide fusion protein comprises an amyloidreactive peptide and an antibody or fragment thereof. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any non-polar amino acid. In some embodiments, y is K. In some83MOFO-360384346.1Attorney Docket No. 165992001640 embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0294] In some embodiments, the method of making an antibody-peptide fusion protein comprises isolating the antibody-peptide fusion protein from the host cells. In some embodiments, the methods comprise isolating the antibody-peptide fusion proteins using Protein A affinity chromatography. In some embodiments, a low pH buffer is used to elute the reagents. In some embodiments, the methods of making an antibody-peptide fusion protein comprises dialyzing isolated products of a Protein A affinity chromatography with phosphate buffered saline (PBS).

[0295] In some embodiments, the method of making an antibody-peptide fusion protein further comprises a continuous cell culture method. In some embodiments, the method comprises culturing cells under perfusion conditions. In some embodiments, the method comprises recovering the antibody-peptide fusion protein about every 12-36 hours. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide and an antibody or fragment thereof. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any nonpolar amino acid. In some embodiments, y is R. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 4. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 8.

[0296] In some embodiments, the method of making an antibody-peptide fusion protein further comprises a continuous cell culture method. In some embodiments, the method comprises culturing cells under perfusion conditions. In some embodiments, the method comprises recovering the antibody-peptide fusion protein about every 12-36 hours. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide and an antibody or fragment thereof. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence as shown in Table 1. In some embodiments, the amyloidreactive peptide comprises an amino acid sequence as shown in Table 1 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions compared84MOFO-360384346.1Attorney Docket No. 165992001640 to the amino acid sequences shown in Table 1. In some embodiments, the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K, and wherein n is at least 2. In some embodiments, x is any nonpolar amino acid. In some embodiments, y is K. In some embodiments, n is 9. In some embodiments, the amyloid-reactive peptide comprises SEQ ID NO: 5.

[0297] In some embodiments, the method of making an antibody-peptide fusion protein comprises culturing a host cell, wherein the host cell is a mammalian cell. In some embodiments, the host cell is a CHO cell. In some embodiments, the host cell comprises a vector encoding an antibody-peptide fusion protein, including any of the antibody-peptide fusion proteins described herein.

[0298] In some embodiments, the method of making an antibody-peptide fusion protein further comprises determining the purity of the antibody-peptide fusion protein. For example, sodium dodecyl sulfate capillary electrophoresis (CE-SDS) is an analytical method used to assess the purity of proteins, including the quantitative analysis of monoclonal antibodies. Antibody samples are mixed with a replaceable SDS-gel buffer and then electrophoresed through an SDS-gel filled capillary. Samples are injected into the capillary inlets capillary using high voltage. Protein migration through the separation matrix occurs in an anodic direction, and quantitative detection occurs near the distal end of the capillary using a UV absorbance detection system. In some embodiments, the purity of the antibody-peptide fusion protein is determined using one or more analytical methods comprising sodium dodecyl sulfate capillary electrophoresis (CE-SDS), liquid chromatography (LC), mass spectrometry (MS), or a combination thereof. In some embodiments, the purity of the antibody-peptide fusion protein is determined using sodium dodecyl sulfate capillary electrophoresis (CE- SDS).

[0299] In some embodiments, the method further comprises determining the purity, wherein the antibody-peptide fusion protein is purified to at least 80% intact antibody-peptide fusion protein. In some embodiments, the antibody-peptide fusion protein has a purity that is defined by the amount of the intact antibody-peptide fusion protein present. In some embodiments, the intact antibody-peptide fusion protein consists of the full length antibody- peptide fusion.

[0300] In some embodiments, the method further comprises determining the purity, wherein the antibody-peptide fusion protein is purified to at least 80% intact antibody-peptide85MOFO-360384346.1Attorney Docket No. 165992001640 fusion protein. In some embodiments, the antibody-peptide fusion protein has a purity that is defined by the amount of the intact antibody-peptide fusion protein present. In some embodiments, the intact antibody-peptide fusion protein consists of the full length antibody- peptide fusion. In some embodiments, the antibody-peptide fusion protein comprises no more than 20% of a cleavage product. In some embodiments, the cleavage product comprises a light chain comprising a light chain lacking one or more amino acid residues from the N- terminus or the C-terminus. In some embodiments, the cleavage product comprises a heavy chain comprising a heavy chain lacking one or more amino acid residues from the N-terminus or the C-terminus.VIII. Kits

[0301] The present disclosure also provides kits containing an amyloid-reactive peptide and / or an antibody-peptide fusion protein of the present disclosure. Kits of the present disclosure may include one or more containers comprising a purified amyloid-reactive peptide and / or a purified antibody-peptide fusion protein. In some embodiments, the kits further include instructions for use in accordance with the methods of this disclosure. In some embodiments, these instructions comprise a description of administration of the antibody- peptide fusion protein as described herein to treat an amyloid-related disorder, according to any methods of this disclosure.

[0302] In some embodiments, the instructions comprise a description of how to detect an amyloid deposit, for example in a subject, in a tissue sample, or in a cell. The kit may further comprise a description of selecting an individual suitable for treatment based on identifying whether that individual has an amyloid disease, as described herein.

[0303] The label or package insert indicates that the composition is used for treating, e.g., a disease of the present disclosure. Instructions may be provided for practicing any of the methods described herein.IX. EXEMPLARY EMBODIMENTS

[0304] Various embodiments provided herein are included in the following non-limiting list of embodiment.

[0305] Embodiment 1. An amyloid-reactive peptide that binds to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K wherein n is at least 2, wherein the amyloid reactive-peptide binds to human amyloid fibrils with a Ec50 that is less than about 1000 nM.86MOFO-360384346.1Attorney Docket No. 165992001640

[0306] Embodiment 2. The amyloid-reactive peptide of embodiment 1, wherein x is selected from a group consisting of A and G.

[0307] Embodiment 3. The amyloid-reactive peptide of embodiment 1 or 2, wherein n is between 2 and 20.

[0308] Embodiment 4. The amyloid-reactive peptide of any of embodiments 1-3, wherein y is R.

[0309] Embodiment 5. The amyloid-reactive peptide of any of embodiments 1-3, wherein y is K.

[0310] Embodiment 6. An amyloid-reactive peptide that binds to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions.

[0311] Embodiment 7. The amyloid- reactive peptide of embodiment 6, wherein the amyloid reactive-peptide binds to human amyloid fibrils with a Ec50 that is less than about1000 nM.

[0312] Embodiment 8. The amyloid-reactive peptide of embodiment 6 or 7, wherein the amino acid substitutions are conservative substitutions.

[0313] Embodiment 9. The amyloid-reactive peptide of any of embodiments 1-8, wherein the amyloid-reactive peptide binds to rVk6WIL fibrils, ATTRwt(PER125) extract, ATTRv(KEN) extract, ALX(SHI) liver extract, ALK(TAL) liver extract, AP(l-40), IAAP, ALK4, ALM, or ATTR fibrils.

[0314] Embodiment 10. The amyloid-reactive peptide of any of embodiments 1-9, wherein the amyloid-reactive peptide binds to A LX, ALK, ATTRV, or rVX6WIL fibrils.

[0315] Embodiment 11. An antibody -peptide fusion protein, comprising: the amyloidreactive peptide of claim any of embodiments 1-10 joined to an antibody or fragment thereof.

[0316] Embodiment 12. The antibody-peptide fusion protein of embodiment 11, wherein the antibody or fragment thereof binds to human amyloid.

[0317] Embodiment 13. The antibody-peptide fusion protein of embodiment 11 or 12, wherein the antibody or fragment thereof binds to human immunoglobulin light chain amyloid.

[0318] Embodiment 14. The antibody-peptide fusion protein of any of embodiments 12- 13, wherein the antibody is a full-length antibody, a Fab fragment, or a scFv.87MOFO-360384346.1Attorney Docket No. 165992001640

[0319] Embodiment 15. The antibody-peptide fusion protein of any of embodiments 11- 14, wherein the antibody or fragment thereof comprises a human Fc region.

[0320] Embodiment 16. The antibody-peptide fusion protein of embodiment 15, wherein the human Fc region is of an IgGl, IgG2, IgG3, or IgG4 isotype.

[0321] Embodiment 17. The antibody-peptide fusion protein of any of embodiments 11- 16, wherein the antibody or fragment thereof comprises a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), wherein: a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19; b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR- H3 comprising the amino acid sequence set forth in SEQ ID NO: 19; or c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR- H3 comprising the amino acid sequence set forth in SEQ ID NO: 19.

[0322] Embodiment 18. The antibody-peptide fusion protein of embodiment 17, wherein the amyloid-reactive peptide and antibody or fragment thereof are joined together at the N-terminal end of the light chain, the C-terminal end of the light chain, the N-terminal end of the heavy chain and / or the C-terminal end of the heavy chain.

[0323] Embodiment 19. The antibody-peptide fusion protein of any of embodiments I lls, wherein the amyloid-reactive peptide and antibody or fragment thereof are joined via a linker.

[0324] Embodiment 20. The antibody-peptide fusion protein of embodiment 19, wherein the linker comprises an amino acid sequence set forth in SEQ ID NO 82.88MOFO-360384346.1Attorney Docket No. 165992001640

[0325] Embodiment 21. The antibody-peptide fusion protein of any of embodiments 11- 20, wherein the amyloid-reactive peptide and antibody or fragment thereof comprise an N- terminal spacer.

[0326] Embodiment 22. The antibody-peptide fusion protein of embodiment 21, wherein the N-terminal spacer comprises an amino acid sequence set forth in SEQ ID NO: 83 or SEQ ID NO: 103.

[0327] Embodiment 23. The antibody-peptide fusion protein of any of embodiments 11- 22, wherein the amyloid-reactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and the antibody-peptide fusion protein comprises a secretory leader sequence.

[0328] Embodiment 24. The antibody-peptide fusion protein of embodiment 23, wherein the secretory leader sequence comprises an amino acid sequence set forth in SEQ ID NO: 84.

[0329] Embodiment 25. The antibody-peptide fusion protein of any of embodiments 11-24, wherein the antibody or functional fragment thereof comprises human framework sequences.

[0330] Embodiment 26. The antibody-peptide fusion protein of any of embodiments 11-25, wherein the antibody is humanized.

[0331] Embodiment 27. The antibody-peptide fusion protein of any of embodiments 17-26, wherein the VL comprises one or more amino acid residues selected from the group consisting of: a. Tyr at position 36; b. Leu at position 37; c. Leu at position 46; d. Leu at position 85; and e. Phe at position 87, wherein the VH comprises one or more amino acid residues selected from the group consisting of: a. Vai at position 37; b. Leu at position 48; c. Leu at position 67; d. Ser at position 68; e. Lys at position 71; f. Ser at position 76; g. Vai at position 78; h. Leu at position 79; i. Phe at position 80; j. Thr at position 89; k. Vai at position 93; and 1. Thr at position 94, wherein the amino acid positions are numbered according to the numbering system of Kabat.

[0332] Embodiment 28. The antibody-peptide fusion protein of embodiment 27, wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises Leu at position 48, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, and Thr at position 94.

[0333] Embodiment 29. The antibody-peptide fusion protein of embodiment 27, wherein the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a89MOFO-360384346.1Attorney Docket No. 165992001640CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19.

[0334] Embodiment 30. The antibody-peptide fusion protein of embodiment 27, wherein the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32-42.

[0335] Embodiment 31. The antibody-peptide fusion protein of embodiment 27 or 30, wherein the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63.

[0336] Embodiment 32. The antibody-peptide fusion protein of embodiment 27, wherein the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO:55.

[0337] Embodiment 33. The antibody-peptide fusion protein of embodiment 27, wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:34, and the VH comprises an amino acid sequence set forth in SEQ ID NO:48.

[0338] Embodiment 34. The antibody-peptide fusion protein of any of embodiments 11- 33, wherein the amyloid-reactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and wherein the light chain with the amyloidreactive peptide fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 86, 87, and 93-97.

[0339] Embodiment 35. The antibody-peptide fusion protein of embodiment 34, wherein the light chain with the amyloid-reactive peptide fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NO: 93 or 86.

[0340] Embodiment 36. The antibody-peptide fusion protein of any of embodiments 11- 35, wherein the amyloid-reactive peptide and antibody or fragment thereof are joined together at the N-terminal end of the light chain and wherein the light chain with the amyloidreactive peptide fused to the N- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 85, 89, 91, and 92.

[0341] Embodiment 37. The amyloid-reactive peptide of any of embodiments 1-10, or the antibody-peptide fusion of embodiments 9-34, wherein the amyloid-reactive peptide is conjugated to a detectable label.90MOFO-360384346.1Attorney Docket No. 165992001640

[0342] Embodiment 38. The amyloid-reactive peptide or antibody -peptide fusion of embodiment 37, wherein the detectable label is selected from the group consisting ofnC,18F,i nIn, "mTc,89Ga,123I,124I,125I, a biotin and a fluorophore.

[0343] Embodiment 39. The amyloid-reactive peptide or antibody-peptide fusion of embodiment 37 or 38, wherein the detectable label is124I.

[0344] Embodiment 40. The amyloid-reactive peptide or antibody-peptide fusion of embodiment 37 or 38, wherein the detectable label is "mTc.

[0345] Embodiment 41. A pharmaceutical composition comprising the amyloid reactive peptide of any one of embodiments 1-8 and 35-38 or the amyloid-peptide fusion protein of any one of embodiments 11-39.

[0346] Embodiment 42. The pharmaceutical composition of embodiment 41, further comprising a pharmaceutical acceptable carrier.

[0347] Embodiment 43. The pharmaceutical composition of any of embodiments 40-42, wherein the pharmaceutical composition is formulated for intravenous or subcutaneous administration.

[0348] Embodiment 44. Nucleic acid(s) encoding the amyloid reactive peptide of any one of embodiments 1-10 and 35-38 or the amyloid-peptide fusion protein of any one of embodiments 11-40.

[0349] Embodiment 45. A vector comprising the nucleic acid(s) of embodiment 44.

[0350] Embodiment 46. A host cell comprising the vector of embodiment 45.

[0351] Embodiment 47. The host cell of embodiment 46, wherein the host cell is a mammalian cell, optionally a Chinese hamster ovary (CHO) cell.

[0352] Embodiment 48. A method of making an antibody-peptide fusion comprising culturing the host cell of embodiment 47 under conditions suitable for expression of the vector encoding the antibody-peptide fusion protein and recovering the antibody-peptide fusion protein.

[0353] Embodiment 49. A method of treating a subject having an amyloid-related disorder comprising an amyloid deposit, comprising administering to the subject an effective amount of the antibody-peptide fusion protein of any one of embodiments 11-40.

[0354] Embodiment 50. The method of embodiment 49, wherein treating the subject with the antibody-peptide fusion protein causes phagocytosis of the amyloid deposit, causes clearance of the amyloid deposit and or prevents formation of additional amyloid deposits.91MOFO-360384346.1Attorney Docket No. 165992001640

[0355] Embodiment 51. A method of identifying an amyloid deposit in a subj ect, comprising administering the amyloid-reactive peptide or the antibody-peptide fusion protein of embodiments 37-40 to the subject, and detecting a signal from the detectable label.

[0356] Embodiment 52. The method of embodiment 51, further comprising diagnosing the subject with amyloid-related disorder if a signal is detected.

[0357] Embodiment 53. The method of embodiment 51 or 52, further comprising treating the subject for an amyloid-related disorder if a signal is detected.

[0358] Embodiment 54. The method of any of embodiments 49, 50, 52 and 53, wherein the amyloid-related disorder is systemic amyloidosis or localized amyloidosis.

[0359] Embodiment 55. The method of any of embodiments 49, 51, and 52-54 wherein the amyloid-related disorder is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, APrP, and Ap amyloidosis.

[0360] Embodiment 56. The method of any of embodiments 49, 50, and 52-55, wherein the amyloid-related disorder is systemic amyloidosis.

[0361] Embodiment 57. The method of any of embodiments 49, 50, and 52-56, wherein the amyloid- related disorder is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, and ACys,

[0362] Embodiment 58. The method of any of embodiments 49, 50, and 52- 57, wherein the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

[0363] Embodiment 59. The method of any of embodiments 49, 50, and 53-58, wherein the amyloid deposit is opsonized by the antibody-peptide fusion protein.

[0364] Embodiment 60. The method of any of embodiments 49-59, wherein the subject is a human.X. EXAMPLES

[0365] The following examples further illustrate the invention but should not be construed as in any way limiting its scope. In light of the present disclosure and the general level of skill in the art, those of skill will appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications, and alterations can be employed without departing from the scope of the presently disclosed subject matter. The attached figures are meant to be considered as integral parts of the specification and description of the disclosure.92MOFO-360384346.1Attorney Docket No. 165992001640Example 1. Design of amyloid-reactive peptides

[0366] Amyloid-reactive peptides pl 08 (SEQ ID NO: 1), pl 09 (SEQ ID NO: 2), pl 10 (SEQ ID NO: 3), pl 12 (SEQ ID NO: 4), pl 13 (SEQ ID NO:5) and pl 14 (SEQ ID NO: 6) were chemically synthesized and received as lyophilized material with greater than 85% purity. In addition, amyloid-reactive peptides pl08 (SEQ ID NO: 1), pl09 (SEQ ID NO: 2), pl 10 (SEQ ID NO: 3) were synthesized with a biotinylated N-terminal to allow detection enzyme linked immunosorbent assay (ELISA) and immunohistochemical studies.Table 4: Exemplary amyloid-reactive peptide sequenceExample 2. Structural determination by Circular dichroism for amyloid-reactive peptides

[0367] Structural determination of peptide pl 12 was assessed using circular dichroism (CD) in the absence and presence of increasing concentrations of low molecular weight heparin (enoxaparin).

[0368] Circular dichroism (CD) spectra of peptide pl 12 (0.05 mg / mL in PBS) were acquired using a DSM 1000 CD instrument (Olis Inc., Bogart, Georgia) with a peptide sample volume of 3 mL and a 1 cm cuvette path length. Data were collected in triplicate over the 200 nm - 250 nm wavelength range with 1 nm increments. The effect of enoxaparin on the secondary structure was assessed by addition of enoxaparin up to 66 pg / mL. Spectra93MOFO-360384346.1Attorney Docket No. 165992001640 were corrected for background by subtraction of a PBS-only CD spectrum. Mean residue ellipticity [9] was calculated according t Equation

[0369] In Equation 1, 9 is elipticity (millidegrees); MW is the molecular weight of the peptide; no. AA, is the number of amino acid residues; cone, is the peptide concentration (mg / mL), and; 1, is the cuvette pathlength (cm).

[0370] As shown in FIG. 1A-1C, the CD spectrum of peptide pl 12 in PBS differs dramatically from that seen for classical a-helical proteins and peptide (see e.g. the CD spectrum of bovine serum albumin (BSA) (FIG. 1C)). The spectrum is reminiscent of the polyproline coil structure adopted by collagen, Lopes, J.L., et al., Distinct circular dichroism spectroscopic signatures of polyproline II and unordered secondary structures: applications in secondary structure analyses. Protein Sci, 2014. 23(12): p. 1765-72. Addition of enoxaparin, from 13 pg / mL to 66 pg / mL did not significantly alter the secondary structure of pl 12 which demonstrated the rigidity of the peptide (FIG. 1A). This differed significantly from the enoxaparin-induced a-helix formation seen with other polybasic pan-amyloid binding peptides, such as peptide p5+14 (SEQ ID NO: 12) (FIG. IB). These data suggested that the polybasic peptide pl 12 is not an a-helical peptide and differs from other helical and P-sheet amyloid-reactive peptides.Example 3: In vitro bioactivity assays for amyloid-reactive peptides rV 6WIL fibril pulldown assay

[0371] Binding of radioiodinated biotinylated pl 08, pl 09, and pl 10 and non-biotinylated pl 12 with synthetic rV+6WIL fibrils was assessed in a pulldown assay. The binding of radioiodinated peptide p5 (+8 charge) and p5+14 (+12 charge) served as positive controls.

[0372] Synthetic amyloid like fibrils were prepared from a recombinant immunoglobulin +6 light chain variable domain (rV+6WIL) and used as the substrate to assess the binding of radioiodinated peptides and peptide fusions.

[0373] Methods'. Peptides biotinyl-pl08, -pl09, -pl 10 as well as non-biotinylated-pl 12 (100 pg) and p5 and p5+14 (as positive controls) were radioiodinated with ~2 mCi of iodine- 125 (Perkin Elmer, Waltham, MA, USA), using 20 pg chloramine T as the oxidizing agent. After quenching the reaction with 20 pg sodium metabisulfite, the radiolabeled peptide was diluted into 0.1% sterile gelatin in PBS and free radioiodide removed by size exclusion chromatography on a 5 mL PD10 (GE Healthcare) solid phase equilibrated with 0.1%94MOFO-360384346.1Attorney Docket No. 165992001640 gelatin / PBS. Fractions of 200 pL were collected and those containing the maximal radioactivity (indicative of125I-labeled peptide) were pooled, and the product’s radiochemical purity was established by SDS polyacrylamide gel electrophoresis (PAGE) analyzed by phosphor imaging (Cyclone Storage Phosphor System, Perkin Elmer, Shelton, CT, USA).

[0374] Binding to the synthetic amyloid-like fibrils was performed using, as the substrate, 25 pg of rVk6WIL fibrils suspended in PBS with 0.05 % (v / v) Tween 20. The fibrils were pelleted by centrifugation at 13,000 x g for 3 min before being resuspended in 200 pL PBS.

[0375] The radiolabeled peptides (~10 ng and ~ 100,000 cpm activity) was added to the fibril suspension and the tubes incubated at RT with end over end mixing for 1 hour. The amyloid and any bound125I-peptide was isolated by centrifugation and the pellet washed once in a 200 pL volume of PBS.

[0376] The two supernatants obtained following the centrifugation steps were combined in 12 x 75 mm glass tube and served as a measure of the unbound, “free” peptide. The pelleted amyloid was transferred to a separate glass tube by resuspension in 200 pL of PBS and the microcentrifuge tube washed to recover any residue. The 1-125 was quantitated in an automated Wizard 1480 gamma scintillation counter and the percent bound calculated.

[0377] The bioactivity value, % bound, was calculated according using Equation 2.Equation 2: Bioactivity (% bound)= (pellet cpm / (pellot cpm + supernatant cpm)) * 100

[0378] Result'. Peptides pl09, pl 10, and pl 12 bound rVk6WIL fibrils with greater than 95% bound peptide in this assay (Table 5 and FIG. 2). In each case the binding was greater then, or equivalent to the pan-amyloid reactive peptide, p5 and p5+14. Radiolabeled peptide pl 08 bound the amyloid-like fibrils but with a lower percent peptide binding relative to p5, pl 09 and pl 10 (Table 5). p5 is an exemplary a -helical pan-amyloid binding peptide was used as a positive control. These data demonstrated that peptides pl 08, pl 09, pl 10, and pl 12 are capable of binding amyloid fibrils.Table 5: Bioactivity ofI25I-labeled peptides in rV26WIL fibrils in pulldown assay95MOFO-360384346.1Attorney Docket No. 165992001640Amyloid extract binding microplate assay

[0379] Biotinylated peptides pl 09 and pl 10 were assessed for binding to rVk6WIL fibrils as well as human AL and ATTRv amyloid extracts. These extracts contained, in addition to amyloid fibrils, more than 30 accessory proteins that are associated with tissue amyloid in patients. Amyloid extracts are a complex heterogeneous matrix that are more representative of tissue amyloid as compared to synthetic fibrils.

[0380] Methods'. Binding of biotinylated pl 09 and pl 10 to fibrils and amyloid extracts was assessed using a europium -linked immunosorbent assay (ELISA). Briefly, wells of a 96- well polystyrene microplate (Coming, Coming, NY, USA) were coated with amyloid-like fibrils (0.83 pM stock), or human amyloid extracts (0.06 mg / mL), 50 pL of stock preparation per well. Substrate-coated wells were then treated with 200 pL of blocking buffer (PBS containing 1% bovine serum albumin; BSA) for 1 hour at RT before washing with PBS and addition of the appropriate concentrations of peptide in PBS with 1% (w / v) BSA and 0.05% (v / v) tween 20. Following a wash step, the bound peptide was detected by addition of 100 pL of europium-streptavidin (Perkin Elmer, Waltham, MA, USA) and 100 pL of enhancement solution (Perkin Elmer). Time-resolved fluorescence emission was then measured using a Wallac Victor 3 plate reader (Perkin Elmer).

[0381] Both biotinylated pl 09 and pl 10 bound fibrils and amyloid extracts when dried onto the wells of a microplate (FIG. 3A and FIG. 3B). In some cases, a significant prozone effect (excess peptide or Hook effect) was observed at higher concentrations. Binding studies were performed in Superblock (purified glycoprotein blocking agent; ThermoFisher) to minimize the non-specific binding of the charged peptides.

[0382] The binding potency, EC 50, was estimated after fitting the data using a four parameter logistic regression. The EC50 of both peptides for all substrates were remarkably similar, ranging from 3.5 nM - 9.9 nM (Table 6).Table 6: Estimated EC 50 values for the binding of biotinylated peptide pl 09 and pl 10 to rV 6WIL fibrils and human AL andATTR amyloid extracts96MOFO-360384346.1Attorney Docket No. 165992001640

[0383] In a similar ELISA based study, APF2 and APF4, were shown to bind human ALA. amyloid extract dried onto the wells of a microplate (FIG 4A and FIG. 4B). The binding potency (EC50) was estimated after fitting the data and found to be 0.7 nM for APF4 and 2.9 nM APF2 for rVA6WIL fibrils, and 1.3 nM for APF4 and 3.2 nM for APF2 for ALA extract.

[0384] These data suggested that the novel amyloid reactive peptides, either alone or when fused to an Ig light chain in the context of an IgG, are capable of high potency binding to amyloid fibrils and human amyloid extracts.Example 4. In vivo binding and activity assays for amyloid-reactive peptidesIn vivo binding of amyloid-reactive peptides with mouse AA

[0385] Methods: Peptides biotinyl-pl08, biotinyl -pl 09, biotinyl -pl 10 and nonbiotinylated pl 12 (100 pg) were radioiodinated as described above. Transgenic H2-Ld-huIL-6 Balb / c mice (H2 / IL-6) that constitutively expressed the human IL-6 transgene, were treated for induction of systemic serum amyloid protein A (AA) associated amyloidosis. Briefly, 10 pg of amyloid enhancing factor (AEF), isolated by water flotation from spleens of diseased mice, was injected intravenously (IV) in 100 pL of PBS into huIL-6 transgenic 8-12 week old mice. Male and female mice were used 4-6 week post- AEF injection and amyloid-free, strain and age-matched wild type (WT) mice served as the negative control.

[0386] For SPECT / CT imaging radioiodinated peptides (-200 pCi 1-125 and -10 pg of peptide) were injected IV in 200 pL of PBS in the lateral tail vein of the mice. Animals were euthanized at 1 h or 4 h post injection by isoflurane inhalation overdose and SPECT / CT images acquired (described below) followed by necropsy. Tissues and a blood sample were harvested from each animal, weighed in tared vials, and radioactivity quantified using a Wizard 1470 gamma spectrometer (Perkin Elmer). The % injected dose per gram (%ID / g) of tissue was calculated.

[0387] Image data were collected using an Inveon Trimodality SPECT / PET / CT instrument (Siemens Preclinical, Knoxville, TN). SPECT images were generated by acquiring sixty 16 second projections per revolution using 56 mm of bed travel in 1.5 revolutions and a 6-degree angle between projections. A 1.0-mm-diameter five-pinhole (Mouse Whole Body) collimator was used at 30 mm from the center of the field of view. Data were reconstructed using a Point Spread Function (PSF) model and a MAP 3D algorithm with (16 iterations and 6 subsets) with a MAP prior of 1.000 using attenuation correction and scatter correction.97MOFO-360384346.1Attorney Docket No. 165992001640

[0388] CT data were acquired using an x-ray voltage biased to 80 kVp with a 500uA anode current with an aluminum filter with 0.5 mm thickness. A 200 ms exposure was used and 361 projections were collected covering 360 degrees of rotation on low setting with a Dalsa CT camera. The data were reconstructed using an implementation of the Feldkamp- filtered back-projection algorithm, using beam hardening correction onto a 512 x 512 x 608 matrix. The data were then down-sampled by a bin of 4. An ROI (region of interest) was drawn over the heart using IRW (Inveon Research Workplace, Siemens Molecular Imaging). The time of scan was noted for decay correction. The volume, mean, standard deviation, and max pixel values were recorded.

[0389] Results: All three peptides colocalized with amyloid deposits in the heart, kidney, liver and spleen as evidenced by microautoradiography (FIG. 5A-5C). Uptake of pl 09 and pl 10 in the cardiac amyloid deposits was particularly intense. In addition to this, amyloid in the renal glomeruli, perivascular and sinusoidal deposits in the liver, and amyloid in the perifollicular regions of the spleen also showed peptide accumulation. The dense waxy amyloid deposits in the spleen were characterized by strong peptide binding at the periphery of the amyloid but little or no accumulation within the dense waxy mass (20x images of pl 09 and pl 10, FIG. 5B and FIG. 5C). This study indicated that biotinylated peptides pl 08, pl 09 and pl 10 bind systemic AA amyloid deposits in vivo.

[0390] The biodistribution of125I-labeled peptide pl 12 was also assessed in mice with systemic AA amyloid and in WT control mice, at 1 hour and 4 hours post injection. Tissue binding was measured as %ID / g of tissue. At 1 hour post injection, the uptake of125I-p 112 in the liver, spleen and kidney was 5-10 %ID / g. In the pancreas uptake was ~20 %ID / g, the stomach was similarly high (FIG. 6A). In contrast, the uptake all organs of the WT mice was less than ~2 %ID / g. At 4 hours post-injection, the peptide uptake in all organs was decreased with ~5 %ID / g in the liver, pancreas and spleen, and less in the kidney. In contrast, the stomach radioactivity remained high at ~20 %ID / g, which may be indicative of dehalogenation of the peptide and accumulation of free radioiodide in the stomach lumen (FIG. 6B)

[0391] These data indicated that peptide pl 12 accumulated in amyloid laden organs in the mice with AA amyloidosis, but not in amyloid-free organs of a WT mouse. Peptide was either released from the amyloid between 1 hour and 4 hour post injection, or was dehalogenated by enzymatic activity, resulting in loss of tissue radioactivity at the later time point.98MOFO-360384346.1Attorney Docket No. 165992001640Immunohistochemical (IHC) evaluation of tissue amyloid binding

[0392] The binding of biotinylated peptides p5, pl 09 and pl 10 to human AL and ATTRv amyloid in formalin-fixed, paraffin-embedded tissue sections was assessed by immunohi stochemi stry .

[0393] Methods: Tissue was collected and after routine histology procedure for deparaffinizing tissue slides, from Clearing Agent, through various alcohols the slides were washed in tap water. The slides were placed in Dako (SI 699) Target Retrieval Solution at 100°C for 30 minutes. Thereafter the slides were allowed to cool for 20 minutes room temperature before being washed 3 x 5 minutes in PBS (Fisher DPPBS SH130013.03 HyClone). The slides were then placed in hydrogen peroxide, 3% Solution (Fisher S25359) for 12 minutes at room temperature (RT) before being washed 3 x 5 minutes in PBS. Avidin and biotin blocking was performed at RT according to the manufacturer’s instructions (Vector SP-2001). This was followed by a 3 x 5 minute wash in PBS. From stock solutions of biotinylated pl09, pl 10, p5, (0.5mg / ml) a 1 :300 dilution was made in PBS and added to the slides and they were incubated overnight at RT in a humid chamber to prevent evaporation. The slides were then washed in tris buffered saline with Tween 20 (TBST; Dako S3306) for 30 minutes at RT before addition of Vector Laboratories (Vectastain PK-6101) Elite ABC Kit-Peroxidase and incubation for 1 hour at RT. Slides were then washed for 3 x 5 minutes in TBST before being stained by addition of ImmPACT DAB Substrate Kit-Peroxidase solution (Vector SK-4015) with incubation at RT for ~4 minutes Slides were gently washed for 5 minutes under running tap water before staining with Gill #3 hematoxylin (Sigma GHS 3128) using 5 brief immersions in the solution. Slides were again gently washed for 5 minutes under running tap water before taking them through graded alcohols and finally Clearing Agent followed by addition of permanent coverslip medium. Cover slipped slides were air-dried.

[0394] The binding of biotinylated peptide pl 09 and pl 10 were assessed for their ability to specifically bind human AL and ATTRv tissue amyloid in formalin-fixed tissue sections (FIG. 7A-7J).

[0395] Both pl 09 and pl 10 peptides bound the amyloid in the tissue sections obtained from diverse organs. Qualitatively, the binding of both peptides was stronger (based on the intensity of the brown DAB staining) than the reactivity of the positive control, biotinylated peptide p5.

[0396] These data supported the reactivity of peptide pl 09 and pl 10 with human amyloid and, in addition to the data presented above, support the fact that these peptides may exhibit pan-amyloid reactivity.99MOFO-360384346.1Attorney Docket No. 165992001640Example 5: Design and synthesis of antibody-peptide fusion proteins

[0397] Exemplary antibody-peptide fusion proteins were generated. Plasmids comprising the light chains of antibody peptide fusion protein 6 (APF6), APF2, APF4 were synthesized. The plasmids were transiently expressed along with human IgGl heavy chains in HEK or CHO cells.

[0398] In a plasmid expressing the human immunoglobulin heavy chain domains CH2 and CH3 (Fc domain) were generated with the heavy chain of APF8.

[0399] APF2, APF4, APF6 and APF8 were generated by transient expression in HEK or CHO cells. Cells were grown in IgG-depleted medium. Following 6 days in tissue culture, cells were pelleted and the culture medium clarified by centrifugation at 4000 x g. The antibody-peptide fusion proteins were isolated by Protein A affinity chromatography and elution of the reagents in low pH buffer. Isolated products were dialyzed extensively against phosphate buffered saline and stored for use in solution at 4°C.

[0400] The sequences for the components of the antibody-peptide fusion proteins are shown in Table 7.Table 7: Exemplary Antibody Peptide Fusion Proteins:100MOFO-360384346.1Attorney Docket No. 165992001640101MOFO-360384346.1Attorney Docket No. 165992001640Example 6. In vitro bioactivity assays for antibody-peptide fusion proteinsHeparin binding microplate assay

[0401] Pan-amyloid reactive peptides have been shown to bind both amyloid fibrils and hypersulfated heparan sulfate glycosaminoglycans (hHSPG), which are ubiquitous components of amyloid. The low molecular weight heparin, enoxaparin, served as a surrogate for hHSPG. The bioactivity of the peptides and peptide fusions were assessed using an enoxaparin-binding assay.

[0402] Methods: A standard high-binding ELISA plate (Costar 3369) was coated with 100 pl / well of poly-L-lysine (Sigma) at 10-20 pg / ml in PBS, overnight at 4°C in a humid chamber. A poly-L-lysine solution was removed and the plate was washed once with 200 pl PBS. 100 pl / well enoxaparin at 100 pg / ml per well were added, and incubated 1 hour at room temperature (RT). The wells were emptied and washed with PBS. The plate was blocked by adding 200 pl / well of PBS- 1% (w / v) bovine serum albumin (BSA) for 1 hour at RT. The immunoglobulin and Fc-peptide fusions were diluted to the desired starting concentration102MOFO-360384346.1Attorney Docket No. 165992001640(generally 300 nM) in assay buffer (PBS with 1% (w / v) BSA and 0.1% (v / v) Tween20 [PBST]). The blocking solution was removed by inverting the plate and then filling the wells with 100 pl of assay buffer. The diluted fusion proteins were added to the empty starting row (150 pl) and three-fold serial dilutions were performed by transferring 50 pl down the plate. The plates were incubated for 1 hour at RT and the contents were emptied and washed 2 times with PBST. 100 pl / well of biotin-labeled goat anti-human IgG Fey (Jackson Labs) at a dilution of 1 :4000 was added and the plates were incubated 1 hour at RT. The plates were emptied by inversion and washed twice with PBST. 100 pl / well, europium-labeled streptavidin (Perkin-Elmer, 1 / 2000 dilution) was added to the plate and it was incubated for 1 hour at RT. The plate was emptied and washed three times with PBST. 100 pl / well Enhancing solution (Perkin-Elmer) was added and after incubation, the time-resolved fluorescence emission was measured using the Wallac Victor3 plate reader (Perkin-Elmer).

[0403] Results'. Both APF4 and APF2 bound heparin with saturable binding (FIG 8). The APF2, with the peptide placed at the N-terminal of the Ig light chain, bound with greater potency as compared to the Ig-peptide fusion with the peptide placed at the light chain C- terminal. The binding of both pl 12 peptide fusions bound enoxaparin with similar potency to that of antibody-peptide fusion 1 (APF1). The estimated EC50 values for APF4 and APF2 and APF1 binding to enoxaparin in this assay were 1.2 nM, 0.3 nM and 0.7 nM, respectively.

[0404] The reactivity of the Fc-peptide fusion (APF8) and APF6 reagents were assessed in a similar enoxaparin binding assay (FIG. 9). Both reagents bound enoxaparin in this assay, but with moderately lower potency as compared to APF1. The estimated EC50 values for the binding APF8 and APF6 and APF1 to heparin in this assay were 5.0 nM, -21.0 nM and 1.1 nM, respectively.

[0405] These data indicates that the Ig-peptide fusion incorporating peptide pl 12 at either the N- or C-terminal for the light chain is bioactive and capable of binding hHSPG. Incorporating the peptide at the N-terminal resulted in modestly greater potency of binding to enoxaparin. Similarly, the bioactivity of peptide pl 14 was retained and bound hHSPG when incorporated at the C-terminal of an Ig light chain or Fc domain. Peptide pl 12 Ig fusions (e.g., APF2 and APF4) had higher binding potency relative to the peptide pl 14-Ig fusions (e.g., APF6 and APF8).Example 7. Ex-vivo phagocytosis

[0406] Immunoglobulin- and Fc-peptide fusion proteins can serve as therapeutic agents for the clearance of tissue amyloid in patients with systemic amyloidosis. Following binding103MOFO-360384346.1Attorney Docket No. 165992001640 of the reagent to the amyloid through interactions with the amyloid-reactive peptide, the Ig Fc domain can stimulate the innate immune system and induce uptake (phagocytosis) of the amyloid by tissue macrophages or tissue-infiltrating peripheral blood monocytes.

[0407] Methods: Human THP-1 monocytes (~lxl06 / well) were plated in 24-well tissue culture-treated plates. Phorbol-12-myristate-13-acetate (PMA) was added at a final concentration of 50 ng / ml and the cells were allowed to differentiate for 24 hours. After 24 hours, media containing PMA was removed and replaced with fresh media and the cells were allowed to ‘rest’ for 48 hours.

[0408] A phagocytosis assay was performed. The cells were washed with DPBS and 1 ml of RPMI medium was added to each well. The peptide fusions (final concentration of 10 nM) and pHrodo red-labelled rV / AWIL fibrils or human amyloid AL extract (final concentration of 20 pg / ml) were added to each well and the plate was incubated at 37°C for 1 hour to enable phagocytosis. The amount of phagocytosis was quantified by monitoring the increased fluorescence emission of the pHrodo red as the dye enters the acidified phagolysosome, as assessed by fluorescence microscopy (Keyence BZ X800 V 1.3.1). The amount of fluorescence in each image was quantified using image segmentation (Image Pro Premier V 9.0).

[0409] Human THP-1 cells were activated with PMA to M0 macrophages and exposed to pHRodo red-labeled rV / AWIL fibrils and ALA. amyloid extract, in the presence of 10 nM APF8 or APF6. A non-specific human Fc (hFCl) and human IgGl (hlgGl) served as negative controls. Opsonization of the amyloid by the peptide-fusions resulted in a significant enhancement of the phagocytosis of both the fibrils and the amyloid extract (FIG. 10).

[0410] A similar enhancement of amyloid phagocytosis by activated THP-1 cells was observed when APF2 was used as the opsonin (FIG. 11). In this case the uptake of synthetic rV / AWIL fibrils was significantly greater than that seen for the ALL amyloid extract.

[0411] These data indicate that binding of APF2, APF6, and APF8 to amyloid can induce phagocytosis of the amyloid substrates by macrophages, which is considered to be a critical step in the cell-mediated clearance of amyloid in response to immunotherapy.Example 8. Function and Stability of antibody-peptide fusion proteins compared to antibody-peptide fusion 1 (APF1)

[0412] The stability of antibody-peptide fusion proteins comprising antibody reactive peptides, p 112 (SEQ ID NO : 4), p 114(60- 10) (SEQ ID NO : 7), and p 115 (SEQ ID NO : 8) were tested. The peptides were attached to each N-termini of the light chains as shown in104MOFO-360384346.1Attorney Docket No. 165992001640FIG. 12A or the C-termini of the light chain as shown in FIG. 12B. APF1 was used as a comparison. APF3, APF5, APF9, APF11, APF6, and APF7 were generating according to the methods described for generating antibody-peptide fusion proteins in example 5. The sequences of the antibody-peptide fusion proteins are shown in Table 7. APF1 was generated as a control.

[0413] APF3, APF5, APF7, APF9, and APF11 comprise an additional Tyrosine amino acid compared to counterparts with the same sequences otherwise. The tyrosine is a hydrophobic amino acid that was included to improve detection of the amyloid-reactive peptide with mass spectrometry and generation of the antibody-peptide fusion protein.Ex vivo mouse serum stability

[0414] Mouse serum stability assays were performed using heparin-plate binding as a measure of antibody bioactivity. Heparin binding assays were performed according to the methods described in Example 6. The antibodies were incubated in mouse serum at 37°C. The Bioactivity of the reagents was assessed using a heparin binding assay (ELISA using heparin-coated plates).

[0415] As shown in FIG. 13, APF9, APF3, APF5 and APF6 exhibited improved serum stability related to APF1. APF11 degraded before addition of the antibody-peptide fusion to the assay.Binding potency

[0416] Binding of the antibody peptide fusion proteins, APF3, APF9, APF5, APF7 to human amyloid extracts and synthetic amyloid fibrils was tested. The binding potency was compared to binding of APF1 as a control.

[0417] The antibody-peptide fusion proteins were incubated with the human amyloid extracts and the amyloid fibrils and the binding was assessing using an ELISA assay. Gelatin was used as the blocking agent. As shown in FIG. 14, all of the tested antibody-peptide fusion proteins exhibited similar binding potency to antibody-peptide fusion 1. APF11 degraded before addition to the assay.

[0418] Binding of the antibody-peptide fusion proteins was also tested for neural fibrils. The antibody-peptide fusion proteins were incubated with the amyloid fibrils. The binding was assessed using an ELISA assay. Gelatin was used as the blocking agent. As shown in FIG. 15A, APF3 and APF9 bound to Tau fibrils. As shown in FIG. 15B, APF9, APF3, and APF5 bound to Synuclein fibrils.105MOFO-360384346.1Attorney Docket No. 165992001640Phagocytosis

[0419] Phagocytosis assays were performed using pHrodo red-labled amyloid or fibrils according to the methods described in Example 5. The phagocytosis of rVZWIL fibrils, ALA(BAL) and ATTRwt by APF6, APF5, APF3, APF9, APF11, APF13, and APF14 was compared to phagocytosis of rVZWIL fibrils, ALk(BAL) and ATTRwt by APF1. (FIGs. 16A-16C). APF13 comprises a PAR construct where the arginine and alanine residues were reversed in the peptide and containing a tyrosine and C terminus peptide (SEQ ID NO: 101). APF14 comprises a PGR construct where the arginine and glycine residues were reversed in the peptide and containing a tyrosine and C terminus peptide (SEQ ID NO: 102). The amyloid fibrils alone (Wil), or treated with an irrelevant human IgGl (hlgGl) were used as negative controls. APF5 performed best against each amyloid or fibril.In-vivo stability in Mice

[0420] The in vivo stability of the antibody-peptide fusion protein was evaluated using early pharmacokinetics (PK) measurements in wild type mice. FIG. 17 shows the level of detection of APF1 and APF15 over time post injection (pi). APF15 comprises four added histidine residues on the light chain following the peptide. The antibody peptide fusion proteins were detected using an anti-IgG antibody (anti-ID B-13f4 Ab). APF1 exhibited a characteristic delayed Tmax (about 4 hours pi) in the base IgG PK profile. In comparison, APF15 did not exhibit a noticeable delayed Tmax in the base IgG PK profile.

[0421] The stability of the peptides alone compared to the base antibodies used in the antibody-peptide fusion proteins were evaluated in wild type Balb / c mice. The antibody- peptide fusion proteins were administered and then separate assays were used to detect the base antibody and the peptide remaining. The APF1 results are shown in FIG. 18A and the APF15 results are shown in FIG. 18B. The large difference in the PK of the APF1 base IgG and APF1 peptide (based on biotin-heparin binding) during the first 48 h post injection was not seen in the APF 15 data.

[0422] The stability of the antibody-peptide fusion proteins was also tested by comparing the percent change in IgG and peptide from 15 minutes pi to 60 hour (FIG. 19A and 19B). This was measured using the change in IgG and heparin bioactivity over time. The values represent the proportion of bioactive peptide relative to IgG. FIG. 19A shows relative stability for APF1 base antibody and APF1 heparin plate binding. FIG. 19B shows relative stability for the base antibody of APF 15 and heparin plate binding. At 24 hours pi the bioactive fraction of APF 15 was 11 times more than APF1 and 37 times more at 48 hours pi.106MOFO-360384346.1Attorney Docket No. 165992001640In-vivo stability in non-human primates

[0423] The in vivo stability of APF1 and APF5 was compared in non-human primates (cynomolgus monkeys). The antibody-peptide fusion proteins were injected intravenously as a slow bolus at a dose of 10 mg / kg and the concentration of the base antibody and peptide were measured. The peptide concentration was measured using biotin-heparin binding. The large difference in the APF1 base IgG and APF1 peptide (based on biotin-heparin binding) during the first 24 h post injection was not seen in the APF5 data. (FIG. 20A and FIG. 20B). The delta between the antibody-peptide fusion and the peptide is smaller in FIG. 20B, indicating enhanced stability.

[0424] The percentage change in base IgG and bioactive IgG was measured for APF1 and APF5. The percentage change compared to 1 hour pi after 2 hours, 8 hours, and 24 hours is shown FIG. 21A and FIG. 21B. These results show a difference between the stability of the APF5 peptide and the APF1. The non-human primate studies are expanded. Groups of 3 male primates are administered antibody-peptide fusion 1 or APF5. Plasma is collected before dosing, 0.25, 1, 2, 8, 24, 48, 96, and 168 hours post dose.EXEMPLARY SEQUENCESAll polynucleotide sequences are depicted in the 5’- 3 direction. All polypeptide sequences are depicted in the N-terminal to C-terminal direction.107MOFO-360384346.1Attorney Docket No. 165992001640108MOFO-360384346.1Attorney Docket No. 165992001640109MOFO-360384346.1Attorney Docket No. 165992001640110MOFO-360384346.1Attorney Docket No. 165992001640111MOFO-360384346.1Attorney Docket No. 165992001640112MOFO-360384346.1Attorney Docket No. 165992001640113MOFO-360384346.1

Claims

Attorney Docket No. 165992001640CLAIMSWe claim:

1. An amyloid-reactive peptide that binds to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amyloid binding motif, (Pxy)n, SEQ ID NO: 9, wherein x is any amino acid, wherein y is R or K wherein n is at least 2, wherein the amyloid reactive-peptide binds to human amyloid fibrils with a Ec50 that is less than about 1000 nM.

2. The amyloid-reactive peptide of claim 1, wherein x is selected from a group consisting of A and G.

3. The amyloid-reactive peptide of claim 1 or 2, wherein n is between 2 and 20.

4. The amyloid-reactive peptide of any of claims 1-3, wherein y is R.

5. The amyloid-reactive peptide of any of claims 1-3, wherein y is K.

6. An amyloid-reactive peptide that binds to human amyloid fibrils, wherein the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-8 or an amino acid sequence with one, two, or three amino acid substitutions, insertions, or deletions.

7. The amyloid- reactive peptide of claim 6, wherein the amyloid reactive-peptide binds to human amyloid fibrils with a Ec50 that is less than about 1000 nM.

8. The amyloid-reactive peptide of claim 6 or 7, wherein the amino acid substitutions are conservative substitutions.

9. The amyloid-reactive peptide of any of claims 1-8, wherein the amyloid-reactive peptide binds to rVX6WIL fibrils, ATTRwt(PER125) extract, ATTRv(KEN) extract, ALX(SHI) liver extract, ALK(TAL) liver extract, AP(l-40), IAAP, ALK4, AL , or ATTR fibrils.114MOFO-360384346.1Attorney Docket No. 16599200164010. The amyloid-reactive peptide of any of claims 1-9, wherein the amyloid-reactive peptide binds to A LA, ALK, ATTRV, or rV / AWIL fibrils.

11. An antibody-peptide fusion protein, comprising: the amyloid-reactive peptide of claim any of claims 1-10 joined to an antibody or fragment thereof.

12. The antibody-peptide fusion protein of claim 11, wherein the antibody or fragment thereof binds to human amyloid.

13. The antibody-peptide fusion protein of claim 11 or 12, wherein the antibody or fragment thereof binds to human immunoglobulin light chain amyloid.

14. The antibody-peptide fusion protein of any of claims 12-13, wherein the antibody is a full-length antibody, a Fab fragment, or a scFv.

15. The antibody-peptide fusion protein of any of claim 11-14, wherein the antibody or fragment thereof comprises a human Fc region.

16. The antibody-peptide fusion protein of claim 15, wherein the human Fc region is of an IgGl, IgG2, IgG3, or IgG4 isotype.

17. The antibody-peptide fusion protein of any of claims 11-16, wherein the antibody or fragment thereof comprises a light chain comprising a light chain variable region (VL) and a heavy chain comprising a heavy chain variable region (VH), wherein: a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19; b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH115MOFO-360384346.1Attorney Docket No. 165992001640 comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19; or c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in any one of SEQ ID NOs:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in any one of SEQ ID NOs:71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19.

18. The antibody -peptide fusion protein of claim 17, wherein the amyloid-reactive peptide and antibody or fragment thereof are joined together at the N-terminal end of the light chain, the C-terminal end of the light chain, the N-terminal end of the heavy chain and / or the C-terminal end of the heavy chain.

19. The antibody-peptide fusion protein of any of claims 11-18, wherein the amyloidreactive peptide and antibody or fragment thereof are joined via a linker.

20. The antibody-peptide fusion protein of claim 19, wherein the linker comprises an amino acid sequence set forth in SEQ ID NO 82.

21. The antibody-peptide fusion protein of any of claims 11-20, wherein the amyloidreactive peptide and antibody or fragment thereof comprise an N-terminal spacer.

22. The antibody-peptide fusion protein of claim 21, wherein the N-terminal spacer comprises an amino acid sequence set forth in SEQ ID NO: 83 or SEQ ID NO: 103.

23. The antibody-peptide fusion protein of any of claims 11-22, wherein the amyloidreactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and the antibody-peptide fusion protein comprises a secretory leader sequence.

24. The antibody-peptide fusion protein of claim 23, wherein the secretory leader sequence comprises an amino acid sequence set forth in SEQ ID NO: 84.116MOFO-360384346.1Attorney Docket No. 16599200164025. The antibody -peptide fusion protein of any of claims 11-24, wherein the antibody or functional fragment thereof comprises human framework sequences.

26. The antibody-peptide fusion protein of any of claims 11-25, wherein the antibody is humanized.

27. The antibody-peptide fusion protein of any of claims 17-26, wherein the VL comprises one or more amino acid residues selected from the group consisting of: a. Tyr at position 36; b. Leu at position 37; c. Leu at position 46; d. Leu at position 85; and e. Phe at position 87 wherein the VH comprises one or more amino acid residues selected from the group consisting of: a. Vai at position 37; b. Leu at position 48; c. Leu at position 67; d. Ser at position 68; e. Lys at position 71; f. Ser at position 76; g. Vai at position 78; h. Leu at position 79; i. Phe at position 80; j . Thr at position 89; k. Vai at position 93; and l. Thr at position 94, wherein the amino acid positions are numbered according to the numbering system of Kabat.

28. The antibody-peptide fusion protein of claim 27, wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises Leu at position 48, Ser at position 76, Vai at position 78, Leu at position 79, Phe at position 80, and Thr at position 94.117MOFO-360384346.1Attorney Docket No. 16599200164029. The antibody-peptide fusion protein of claim 27, wherein the VL comprises a CDR- L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19.

30. The antibody-peptide fusion protein of claim 27, wherein the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32-42.

31. The antibody-peptide fusion protein of claim 27 or 30, wherein the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63.

32. The antibody-peptide fusion protein of claim 27, wherein the VL comprises the amino acid sequence set forth in SEQ ID NO:36 and the VH comprises the amino acid sequence set forth in SEQ ID NO:55.

33. The antibody-peptide fusion protein of claim 27, wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:34, and the VH comprises an amino acid sequence set forth in SEQ ID NO:48.

34. The antibody-peptide fusion protein of any of claims 11-33, wherein the amyloidreactive peptide and antibody or fragment thereof are joined together at the C-terminal end of the light chain and wherein the light chain with the amyloid-reactive peptide fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 86, 87, and 93-97.

35. The antibody-peptide fusion protein of claims 34, wherein the light chain with the amyloid-reactive peptide fused to the C- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NO: 93 or 86.

36. The antibody-peptide fusion protein of any of claims 11-35, wherein the amyloidreactive peptide and antibody or fragment thereof are joined together at the N-terminal end of118MOFO-360384346.1Attorney Docket No. 165992001640 the light chain and wherein the light chain with the amyloid-reactive peptide fused to the N- terminus of the light chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 85, 89, 91, 92.

37. The amyloid-reactive peptide of any of claims 1-10, or the antibody-peptide fusion of claims 9-34, wherein the amyloid-reactive peptide is conjugated to a detectable label.

38. The amyloid-reactive peptide or antibody-peptide fusion of claim 37, wherein the detectable label is selected from the group consisting ofnC,18F,inIn, "mTc,89Ga,1231,124I,125I, a biotin and a fluorophore.

39. The amyloid-reactive peptide or antibody-peptide fusion of claim 37 or 38, wherein the detectable label is124I.

40. The amyloid-reactive peptide or antibody-peptide fusion of claim 37 or 38, wherein the detectable label is "mTc.

41. A pharmaceutical composition comprising the amyloid reactive peptide of any one of claims 1-8 and 35-38 or the amyloid-peptide fusion protein of any one of claims 11-39.

42. The pharmaceutical composition of claim 41, further comprising a pharmaceutical acceptable carrier.

43. The pharmaceutical composition of any of claims 40-42, wherein the pharmaceutical composition is formulated for intravenous or subcutaneous administration.

44. Nucleic acid(s) encoding the amyloid reactive peptide of any one of claims 1-10 and 35-38 or the amyloid-peptide fusion protein of any one of claims 11-40.

45. A vector comprising the nucleic acid(s) of claim 44.

46. A host cell comprising the vector of claim 45.119MOFO-360384346.1Attorney Docket No. 16599200164047. The host cell of claim 46, wherein the host cell is a mammalian cell, optionally a Chinese hamster ovary (CHO) cell.

48. A method of making an antibody-peptide fusion comprising culturing the host cell of claim 47 under conditions suitable for expression of the vector encoding the antibody-peptide fusion protein and recovering the antibody-peptide fusion protein.

49. A method of treating a subject having an amyloid-related disorder comprising an amyloid deposit, comprising administering to the subject an effective amount of the antibody- peptide fusion protein of any one of claims 11-40.

50. The method of claim 49, wherein treating the subject with the antibody-peptide fusion protein causes phagocytosis of the amyloid deposit, causes clearance of the amyloid deposit and or prevents formation of additional amyloid deposits.

51. A method of identifying an amyloid deposit in a subject, comprising administering the amyloid-reactive peptide or the antibody-peptide fusion protein of claims 37-40 to the subject, and detecting a signal from the detectable label.

52. The method of claim 51, further comprising diagnosing the subject with amyloid- related disorder if a signal is detected.

53. The method of claim 51 or 52, further comprising treating the subject for an amyloid- related disorder if a signal is detected.

54. The method of any of claims 49, 50, 52 and 53, wherein the amyloid-related disorder is systemic amyloidosis or localized amyloidosis.

55. The method of any of claims 49, 51, and 52-54 wherein the amyloid-related disorder is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AIAPP, APro, Alns, APrP, and Ap amyloidosis.120MOFO-360384346.1Attorney Docket No. 16599200164056. The method of any of claims 49, 50, and 52-55, wherein the amyloid-related disorder is systemic amyloidosis57. The method of any of claims 49, 50, and 52-56, wherein the amyloid- related disorder is selected from the group consisting of AL, AH, Ap2M, ATTRv, ATTRwt, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, and ACys,58. The method of any of claims 49, 50, and 52- 57, wherein the amyloid-related disorder is AL amyloidosis or ATTR amyloidosis.

59. The method of any of claims 49, 50, and 53-58, wherein the amyloid deposit is opsonized by the antibody-peptide fusion protein.

60. The method of any of claims 49-59, wherein the subject is a human.121MOFO-360384346.1

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