Antibody-peptide fusion proteins for treating amyloid disorders

Abstract

Provided herein are antibody-peptide fusion proteins that include an amyloid-reactive peptide linked to an antibody. Also provided herein are methods of treating an amyloid-based disease by administering the antibody-peptide fusion proteins.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 190,191, filed May 18, 2021, the contents of which are incorporated by reference in their entirety.

[0002] Submitting a sequence listing as an ASCII text file The contents of the following submission in an ASCII text file are incorporated herein by reference in their entirety: Sequence Listing in Computer Readable Form (CRF) (Filename: 165992000640SEQLIST.TXT, Recorded: May 18, 2022, Size: 69,104 bytes).

[0003] The present application relates to antibody-peptide fusion proteins that bind to human amyloid fibrils and methods of using same. [Background technology]

[0004] Amyloidosis is a fatal protein folding disorder characterized by the aggregation and deposition of protein fibrils and heparan sulfate proteoglycans in vital organs and tissues (Merlini, G. et al. (2003) N. Engl. J. Med. 349, 583-596; Merlini, G. et al. (2004) J. Intern. Med. 255, 159-178; De Lorenzi, E. et al. (2004) Curr. Med. Chem. 11, 1065-1084; Merlini, G. (2004) Neth. J. Med. 62, 104-105). The persistent accumulation of amyloid inevitably leads to organ dysfunction and severe morbidity or mortality. The deposits may be cerebral, as in patients with Alzheimer's disease, Huntington's disease, or prion disease, or may be peripheral, as seen in patients with light chain-associated (AL) amyloidosis, transthyretin-associated (ATTR) amyloidosis, and type 2 diabetes. Further subgrouping into localized or systemic indicates whether the precursor protein is produced locally (at the site of deposition) or circulates in the bloodstream and is deposited at a distal anatomical site, respectively (Westermark, P. et al. (2007) Amyloid. 14, 179-183). Although amyloid can affect any organ or tissue, the kidney, pancreas, liver, spleen, nervous tissue, and heart constitute the major sites of deposition in patients with familial or sporadic forms of systemic amyloidosis. Alzheimer's disease currently affects over 4 million Americans, a number that is estimated to increase to over 16 million by 2050. It is the most common form of amyloidosis, generally considered a rare disorder, but is widely underdiagnosed with an estimated over 200,000 cases in the United States.

[0005] Of these, the major peripheral amyloidosis is transthyretin-related (ATTR) amyloidosis, followed by light chain-related (AL) amyloidosis. The former is due to the deposition of wild-type (sporadic) or variant transthyretin (hereditary) and manifests clinically primarily in the peripheral nerves and heart; however, musculoskeletal involvement is common and may precede organ deposition by decades. The latter is a sporadic monoclonal plasma cell dyscrasia that results in the deposition of fibrils composed of immunoglobulin light chain proteins. AL accounts for approximately two-thirds of all peripheral amyloid cases, with a calculated incidence of approximately 1.4 per 100,000 people per year in the United States, comparable to that of acute lymphocytic and chronic myelogenous leukemia (Group, USCSW (2007) United States Cancer Statistics: 1999-2003 Incidence and Mortality Web-Based Report, USDepartment of Health and Human Services Centers for Disease Control and Prevention National Cancer Institute, Atlanta). AL is approximately one-fifth as common as the related plasma cell dyscrasia multiple myeloma, but is arguably more devastating with a median survival of only 13.2 months, due in part to the rapidly progressive nature of organ destruction, the lack of effective anti-amyloid therapeutics, and the inability to effectively diagnose the disease before organ failure occurs. Fewer than 5% of all AL patients survive more than 10 years from the time of diagnosis (Comenzo, RR et al. (2002) Blood 99, 4276-4282). Moreover, in patients with cardiac AL amyloidosis, the median survival is less than 5 months.

[0006] ATTR is a form of systemic amyloidosis. 25% of patients with ATTR amyloidosis die within 24 months of diagnosis. (Gertz and Dispenzieri JAMA 324(1)79-89(2002)). Current therapies do not prevent organ damage. ATTR amyloidosis is caused by transthyretin (TTR) fibrils. Transthyretin is a protein produced by the liver that helps transport thyroid hormone and vitamin A in the blood. Normally, TTR is a tetramer composed of four single-chain monomers. In hereditary ATTR amyloidosis, TTR gene mutations are thought to destabilize the protein and cause tetramer dissociation into monomers that aggregate into amyloid fibrils. In wild-type ATTR amyloidosis, normal TTR protein becomes unstable, misfolds, and forms amyloid fibrils.

[0007] These amyloid fibrils then accumulate in multiple organs throughout the body, such as in the wrist, in narrow passages called the carpal tunnel. This can cause carpal tunnel syndrome, which causes numbness and tingling in the hand and arm. The spinal canal, which can cause narrowing of the spine (spinal stenosis). The heart, which can cause heart failure and / or irregular heart rhythm (called atrial fibrillation).

[0008] Another widespread form of peripheral amyloidosis in the United States is inflammation-associated (AA) amyloidosis, which is associated with chronic inflammatory disorders such as arthritis, tuberculosis, and familial Mediterranean fever. The incidence of AA is highest in certain parts of Europe, with the frequency varying between ethnicities (Buck, F. et al. (1989) Mod. Pathol. 2, 372-377). In areas where familial Mediterranean fever is widespread and untreated, the incidence of AA may be 100%. In Europe, the incidence has been estimated to be 0.86% based on an autopsy study conducted in Denmark (Lofberg, H. et al. (1987) Acta pathologica, microbiologica, et immunologica Scandinavica 95, 297-302); however, in patients with rheumatoid or psoriatic arthritis, the incidence of AA may be as high as 26%. Such a high prevalence may necessitate screening programs to detect the disease earlier. Amyloid deposition is associated with a sustained increase in plasma concentrations of serum amyloid protein A (sAA), the precursor of amyloid fibrils (Rocken, C. et al. (2002) Virchows Arch. 440, 111-122). AA differs from AL in the type of precursor protein it deposits, but shares common mechanistic features with both that are related to fibril formation and deposition (Rocken, C. et al. (2006) J. Pathol. 210, 478-487; Rocken, C. et al. (2001) Am. J. Pathol. 158, 1029-1038).

[0009] In addition to disorders with well-established amyloid pathogenesis, fibrillar deposits with structural and chromatic properties of amyloid have been identified in other syndromes, although their relevance to disease states has yet to be established. For example, in type 2 diabetes, islet amyloid precursor protein (IAPP) is deposited as amyloid in the islets of Langerhans (Jaikaran, ET et al. (2001) Biochim. Biophys. Acta 1537, 179-203). Aggregation of IAPP results in oligomeric structures that are toxic to pancreatic cells (Lin, CY et al. (2007) Diabetes 56, 1324-1332). Thus, it is suggested that the formation of IAPP amyloid in type 1 diabetes patients contributes to β-cell destruction and leads to the transition to insulin dependence (Jaikaran, ET et al. (2001) Biochim. Biophys. Acta 1537, 179-203). In another example, plaques containing amyloid fibrils composed of apolipoprotein AI have been identified in more than half of patients with atherosclerotic carotid arteries (Westermark, P. et al. (1995) Am. J. Pathol. 147, 1186-1192; Mucchiano, GI et al. (2001) J. Pathol. 193, 270-275). Although these fibril deposits are more common in older patients, apo AI is undoubtedly present early in plaque development (Vollmer, E. et al. (1991) Virchows Arch. A. Pathol. Anat. Histopathol. 419, 79-88). As a final example, apo-A-I amyloid has also been identified in menisci obtained from patients having knee replacement surgery and may contribute to the physical deterioration of the joint (Solomon, A. et al. (2006) Arthritis Rheum. 54, 3545-3550).

[0010] In total, over 29 proteins have been identified chemically or serologically as components of fibrils in amyloid deposits. It is the attributes of these proteins that distinguish the disease, determine treatment, and establish prognosis. Although amyloid fibrils are associated with a clinically heterogeneous group of diseases and can form from structurally distinct and functionally diverse precursor proteins, the deposits themselves share many highly similar characteristics, including the occurrence of fibril structures, fibril epitopes, and similar associated molecules, including heparan sulfate proteoglycans (HSPGs). Amyloid is a heterogeneous complex that contains, in addition to fibrils, glycosaminoglycans (GAGs) and especially perlecan HSPG (Ancsin, JB (2003) Amyloid 10, 67-79; Ailles, L. et al. (1993) Lab. Invest. 69, 443-448; Kisilevsky, R. (1994) Mol. Neurobiol. 9, 23-24; Kisilevsky, R. (1990) Lab. Invest. 63, 589-591; Snow, AD et al. (1987) Lab. Invest. 56, 120-123; Li, JP et al. (2005) Proc. Natl. Acad. Sci. USA 102, 6473-6477).

[0011] To date, the most effective therapeutic interventions for removing amyloid deposits, which may promote recovery of organ function and result in improved prognosis, involve the use of amyloid-reactive antibodies as a means of immunotherapy. Several immunotherapies (antibodies) have been developed for amyloid-related diseases, including monoclonal antibody 11-1F4 for the treatment of AL amyloidosis, NEOD001 for patients with AL amyloidosis, GSK2398852 (anti-SAP monoclonal antibody) for amyloidosis, solanezumab for Alzheimer's disease, intravenous IgG (IVIG) for Alzheimer's disease, and bapineuzumab for Alzheimer's disease, as well as aducumumab for Alzheimer's disease. Each of these approaches has limitations or has not met primary outcomes in late-stage clinical trials (phase 2 / 3).

[0012] Thus, there is a need for effective treatments for amyloidosis and amyloid-related diseases. Summary of the Invention

[0013] Provided herein are antibody-peptide fusion proteins that contain an amyloid-reactive peptide linked to an antibody, as well as methods of making and using the same.

[0014] In one aspect, provided herein is an antibody-peptide fusion protein comprising an amyloid-reactive peptide; and an antibody that binds to amyloid fibrils, wherein the antibody comprises a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL), the amyloid-reactive peptide and the antibody are linked at the N-terminus or C-terminus of the heavy chain or light chain, and the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the antibody binds to human amyloid fibrils.

[0015] In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain.

[0016] In some embodiments, the spacer is selected from the group consisting of SEQ ID NO:83 and SEQ ID NO:86.

[0017] In some embodiments, the light chain further comprises a light chain constant region and the heavy chain comprises a heavy chain constant region.

[0018] In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity to any one of the amino acid sequences set forth as SEQ ID NOs:1-13.

[0019] In some embodiments, the antibody-peptide fusion protein comprises two heavy chains and two light chains, with each light chain linked at its C-terminus to an amyloid-reactive peptide.

[0020] In some embodiments, the antibody is a chimeric or humanized antibody.

[0021] In some embodiments, the VL comprises a CDR-L1 comprising the amino acid sequence shown in SEQ ID NO: 64-70, a CDR-L2 comprising the amino acid sequence shown in SEQ ID NO: 21, and a CDR-L3 comprising the amino acid sequence shown in SEQ ID NO: 22, and the VH comprises a CDR-H1 comprising the amino acid sequence shown in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence shown in SEQ ID NO: 18, and a CDR-H3 comprising the amino acid sequence shown in SEQ ID NO: 19. In some embodiments, the VL comprises a CDR-L1 comprising the amino acid sequence shown in SEQ ID NO: 20; a CDR-L2 comprising the amino acid sequence shown in SEQ ID NO: 21, and a CDR-L3 comprising the amino acid sequence shown in SEQ ID NO: 22, and the VH comprises a CDR-H1 comprising the amino acid sequence shown in SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence shown in SEQ ID NO: 71-81; and a CDR-H3 comprising the amino acid sequence shown in SEQ ID NO: 19. In some embodiments, the VL comprises CDR-L1 having the amino acid sequence shown in SEQ ID NO: 64 to 70, CDR-L2 having the amino acid sequence shown in SEQ ID NO: 21, and CDR-L3 having the amino acid sequence shown in SEQ ID NO: 22, and the VH comprises CDR-H1 having the amino acid sequence shown in SEQ ID NO: 17, CDR-H2 having the amino acid sequence shown in SEQ ID NO: 71 to 81, and CDR-H3 having the amino acid sequence shown in SEQ ID NO: 19.

[0022] In some embodiments, 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.

[0023] 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 VL comprises the amino acid sequence set forth in SEQ ID NO:35 and the VH comprises the amino acid sequence set forth in SEQ ID NO:51. 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 VL comprises the amino acid sequence set forth in SEQ ID NO:35 and the VH comprises the amino acid sequence set forth in SEQ ID NO:52. In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:35 and the VH comprises the amino acid sequence set forth in SEQ ID NO:50. In some embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO:35 and the VH comprises the amino acid sequence set forth in SEQ ID NO:49.

[0024] 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.

[0025] In some embodiments, the antibody is a full length antibody. In some embodiments, the antibody comprises an Fc region. In some embodiments, the Fc region is of the IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the antibody is of the IgG1 isotype.

[0026] In another aspect, provided herein is an antibody-peptide fusion protein, the antibody binding to amyloid fibrils, the antibody comprising a first and second polypeptides each comprising a light chain of the antibody, and a third and fourth polypeptides each comprising a heavy chain of the antibody, and an amyloid reactive peptide linked to the N-terminus or C-terminus of the light or heavy chain, the first and second polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 87, and the third and fourth polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 91. In some embodiments, the first and second polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 88, and the third and fourth polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 92. In some embodiments, the first and second polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 89, and the third and fourth polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 91. In some embodiments, the first and second polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 90, and the third and fourth polypeptides comprising the amino acid sequence set forth in SEQ ID NO: 91.

[0027] In another aspect, provided herein is an antibody-peptide fusion protein comprising an amyloid-reactive peptide comprising the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2; and an antibody that binds to human amyloid fibrils, wherein the antibody comprises a variable heavy chain (VH) and a variable light chain (VL), wherein the VH comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19, and the VL comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22; the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, and 83-86.

[0028] In another aspect, provided herein is an antibody-peptide fusion protein comprising an amyloid-reactive peptide comprising the amino acid sequence set forth in SEQ ID NO:2; and an antibody that binds to human amyloid fibrils, wherein the antibody comprises a variable heavy chain (VH) and a variable light chain (VL), wherein the VH comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19, and the VL comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22; the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid-reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence set forth in SEQ ID NO:83.

[0029] In some embodiments, the antibody-peptide fusion protein exhibits an EC50 of less than 1.5 nM for an amyloid substrate.

[0030] In some embodiments, the antibody-peptide fusion protein is conjugated to a detectable label, and the detectable label comprises a fluorescent label or a radioactive label. In some embodiments, the radioactive label is I-123, I-124, F-18, ZR-89, or Tc-99m.

[0031] In some embodiments, the antibody-peptide fusion protein exhibits one or more in vivo characteristics selected from improved biodistribution, pan-amyloid reactivity, and enhanced phagocytosis compared to a reference IgG antibody.

[0032] In some embodiments, the antibody-peptide fusion protein binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, A1λ1, ATTR, α-synuclein, or tau441 fibrils.

[0033] In another aspect, provided herein is a composition, comprising an antibody-peptide fusion protein comprising: i) an amyloid reactive peptide; and ii) an antibody, wherein the antibody comprises a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL), wherein the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and wherein the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer; wherein at least 90% of the antibody-peptide fusion protein is intact. In some embodiments, the antibody is a full-length antibody.

[0034] In some embodiments, the composition comprises 10% or less cleavage products, the cleavage products comprising a VH missing one or more amino acid residues from the N-terminus or C-terminus compared to the amino acid sequence set forth in SEQ ID NO:89 and a VL missing one or more amino acid residues from the N-terminus or C-terminus compared to the amino acid sequence set forth in SEQ ID NO:91.

[0035] In some embodiments, the antibody-peptide fusion protein exhibits an EC50 binding affinity for one or more amyloid substrates, the EC50 binding affinity being less than 1.5 nM.

[0036] In some embodiments, the composition further comprises a pharma- ceutically acceptable carrier.

[0037] In another aspect, provided herein is a polynucleotide encoding an antibody-peptide fusion protein. In another aspect, provided herein is a vector comprising the polynucleotide. In another aspect, provided herein is a host cell comprising the vector. In some embodiments, the host cell is a mammalian cell, optionally a Chinese Hamster Ovary (CHO) cell.

[0038] In another aspect, provided herein is a method of producing an antibody-peptide fusion protein comprising: a) culturing a host cell comprising a vector encoding the antibody-peptide fusion protein under perfusion cell culture conditions suitable for expression of the antibody-peptide fusion protein; and b) harvesting the antibody-peptide fusion protein about every 12-36 hours; the antibody-peptide fusion protein comprises: i) an amyloid reactive peptide; and ii) an antibody, the antibody comprising a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL), the amyloid reactive peptide and the antibody being linked at the C-terminus of the light chain, and the amyloid reactive peptide being linked to the antibody with or without a spacer. In some embodiments, the amyloid reactive peptide is linked to the C-terminus of the constant domain of the antibody light chain.

[0039] In some embodiments, the method further comprises applying the antibody-peptide fusion recovered in step b) to a cation exchange chromatography column and eluting the antibody-peptide fusion protein from the cation exchange chromatography column.

[0040] In some embodiments, the antibody-peptide fusion protein is eluted separately from the truncated antibody-peptide fusion protein.

[0041] In some embodiments, the host cell is a CHO cell.

[0042] In some embodiments, the method further comprises determining the purity of the antibody-peptide fusion protein, wherein the purity of the antibody-peptide fusion protein is determined using one or more analytical methods including sodium dodecyl sulfate capillary electrophoresis (CE-SDS), liquid chromatography (LC), mass spectrometry (MS), or a combination thereof.

[0043] In some embodiments, the antibody-peptide fusion protein is purified to be at least 90% free from intact antibody-peptide fusion protein.

[0044] In another aspect, provided herein is an antibody-peptide fusion protein produced by the methods described herein.

[0045] In some aspects, provided herein is a method for treating a subject with an amyloid-related disorder comprising amyloid deposits, comprising administering a therapeutically effective amount of an antibody-peptide fusion protein to the subject.In some embodiments, the amyloid-related disorder is systemic or localized amyloidosis.In some embodiments, the amyloid-related disorder is selected from the group consisting of AL, AH, Aβ2M, ATTR, transthyretin, AA, AApoAI, AApoAII, AGel, ALys, ALEct2, AFib, ACys, ACal, AMed, AIAPP, APro, AIns, APrP, Parkinson's disease, Alzheimer's disease, or Aβ amyloidosis.

[0046] In some embodiments, the amyloid deposits are opsonized by the antibody-peptide fusion protein. In some embodiments, treating a subject with the antibody-peptide fusion protein causes phagocytosis of the amyloid deposits. In some embodiments, treating a subject with the antibody-peptide fusion protein results in the improvement of one or more clinical features selected from the group consisting of swelling of the lower extremities, severe fatigue, severe weakness, shortness of breath, difficulty breathing, numbness, pain in the hands, wrists, or feet, diarrhea, constipation, unintentional weight loss, tongue enlargement, skin changes, arrhythmia, and difficulty swallowing.

[0047] In another aspect, provided herein is a method of treating a subject having or suspected of having an amyloid-based disease, comprising: a) determining whether the subject has amyloid deposits by i) administering to the subject an antibody-peptide fusion protein, where the antibody-peptide fusion protein comprises a detectable label, and ii) determining whether a signal associated with the detectable label can be detected from the subject; and b) administering an amyloidosis treatment to the subject if a signal is detected.

[0048] In some embodiments, if no signal is detected, the subject is monitored for subsequent growth of amyloid deposits. In some embodiments, the method further comprises determining the intensity of the signal and comparing the signal to a threshold, where if the threshold is exceeded, the subject is determined to bear amyloid deposits. In some embodiments, the antibody-peptide fusion protein is detected by SPECT / CT imaging, PET / CT imaging, gamma scintigraphy, or optical imaging.

[0049] In some embodiments, the amyloidosis treatment comprises administering to the subject an antibody-peptide fusion protein. In some embodiments, administration of the antibody-peptide fusion protein causes phagocytosis of amyloid deposits in the subject. In some embodiments, administration of the antibody-peptide fusion protein results in clearance of amyloid deposits in the subject.

[0050] In another aspect, provided herein is a method of identifying amyloid deposits in a subject comprising administering to a subject an antibody-peptide fusion protein, where the antibody-peptide fusion protein comprises a detectable label, and detecting a signal from the antibody-peptide fusion protein.

[0051] In another aspect, provided herein is a method of monitoring amyloid clearance in a subject comprising contacting an amyloid substrate in the subject with an antibody-peptide fusion protein, wherein the antibody-peptide fusion protein comprises a detectable label, and wherein a peptide of the antibody-peptide fusion protein has binding affinity for the amyloid substrate; and determining a signal from the detectable label, thereby detecting amyloid clearance.

[0052] In some embodiments, the subject is a human.

[0053] In another aspect, provided herein are kits comprising antibody-peptide fusion proteins for use in the methods provided herein. [Brief description of the drawings]

[0054] [Figure 1] FIG. 1 shows a schematic of an antibody-peptide fusion protein with a peptide fused to the N-terminus of the light chain via a short rigid spacer. [Diagram 2] FIG. 1 shows a schematic diagram of an antibody-peptide fusion protein with a peptide fused to the C-terminus of the heavy chain via a short rigid spacer. [Diagram 3] FIG. 1 shows a schematic diagram of an antibody-peptide fusion protein with a peptide fused to the C-terminus of the light chain via a short rigid spacer. [Figure 4] FIG. 1 shows a schematic diagram of an antibody-peptide fusion protein with a peptide fused to the C-terminus of the light chain via a long flexible spacer. [Figure 5A] 1 shows data from europium-linked immunosorbent assays (EuLISA) measuring binding of chimeric (c)11-1F4 and humanized variants, VH10 / VL4, VH9 / VL4, VH8 / VL4, VH7 / VL4, or VH6 / VL3, to synthetic rVλ6Wil light chain amyloid-like fibrils. [Figure 5B]Shown is data from EuLISA measuring binding of VH6 / VL3-p5 (6-3-p5), VH6 / VL3-p5R (6-3-p5R), c11-1F4, or VH6 / VL3 to rVλ6Wil fibrils. [Figure 5C] Shown is data from EuLISA measuring binding of VH9 / VL / 4-p5R to rVλ6Wil fibrils, Per125 wtATTR extracts, Ken ATTR extracts, SHI ALλ liver extracts, or TAL ALκ liver extracts. [Figure 5D] Shown is data from EuLISA measuring binding of VH9 / VL / 4-p5 to rVλ6Wil fibrils, Per125 wtATTR extracts, Ken ATTR extracts, SHI ALλ liver extracts, or TAL ALκ liver extracts. [Figure 5E] Shown is data from EuLISA measuring binding of c11-1F4, m11-1F4, or VH9 / VL4 to rVλ6Wil fibrils. [Figure 5F] Shown is data from EuLISA measuring VH6 / VL3-p5 binding to Sno ATTR extracts (dark grey circles) or Ken ATTR extracts (light grey circles), and c11-1F4 binding to Sno ATTR extracts (black squares). [Figure 5G] Shown is data from EuLISA measuring binding of VH6 / VL3-p5R to Per125 wtATTR (grey circles, see labels), Sno ATTR extracts (dark grey circles), or Ken ATTR extracts (light grey circles), and c11-1F4 binding to Sno ATTR extracts (black squares). Log-transformed molar concentrations (-log(M)) of monoclonal antibodies are shown on the x-axis, and levels of binding (femtomoles europium) are shown on the y-axis. [Figure 6]Figure 1 shows the results of 125I-mIgp5 binding to rVλ6Wil amyloid-like fibrils and human amyloid extracts obtained from tissues in a pull-down assay. The y-axis shows the percentage of 125I-mIgG-p5 bound, and the percentage bound for each sample is shown above the histogram bars. The x-axis shows the types of amyloid extracts tested, including rVλ6Wil fibrils, SNO hereditary (h)ATTR, KEN hATTR, Per125 wtATTR, Per253 wild-type (wt)ATTR, ALκ HIG extract, ALκ TAL extract, ALλ SHI extract, and ALλ TYL extract. Error bars represent standard deviation. [Figure 7A] Figure 1 shows uptake of pHrodo Red-labeled rVλ6Wil fibrils by human THP-1 macrophages alone or in the presence of human (h)IgG control, ch11-1F4, muIgp5 (produced in expiHEK293 cell line), VH6 / VL3-p5, or VH6 / VL3-p5R, as indicated from left to right on the x-axis. The y-axis shows the level of rVλ6Wil fibril uptake (measured in fluorescence units) and error bars represent standard deviation. [Figure 7B] Figure 1 shows phagocytosis of pHrodo Red-labeled rVλ6Wil fibrils by macrophages in the presence of THP-1 alone or in the presence of THP-1 with hIgG control, c11-1F4, mIgp5, VH9 / VL4-p5, or VH9 / VL4-p5R, as indicated from left to right on the x-axis. The y-axis shows the level of phagocytosis (fluorescence units) and error bars represent standard deviation. [Figure 7C] Figure 1 shows phagocytosis of pHrodo Red-labeled rVλ6Wil fibrils by macrophages in the presence of 5 μg Rituxan (chimeric mAb as negative control), 5 μg c11-1F4, 5 μg VH6 / VL3, 5 μg VH9 / VL4, VH6 / VL3-p5R, or VH6 / VL3-p5, as indicated from left to right on the x-axis. The y-axis shows the level of phagocytosis (pHrodo fluorescence) and error bars represent standard deviation. [Figure 8A]FIG. 1 shows the fed-batch and perfusion workflows used to generate intact VH9-D54E / VL4-N33S-VSPSV-p5R. Purity (% intact fusion protein) is shown for each production method. [Figure 8B] 1 shows the results of a binding experiment testing the affinity of fed-batch purified FB.1 and perfusion purified PF.1 for rVλWIL. The left y-axis scale is used for fed-batch purified FB.1 and the right y-axis scale is used for perfusion purified PF.1. [Figure 9A] 1 shows a gel analysis of radiolabeled (125I) PF.1 antibody-peptide fusion protein compared to a radiolabeled (125I) antibody hIgG1 control. Reduced (Reduced) and non-reduced (NR) samples are shown, and the positions of IgG, IgH, and IgL for each protein are indicated. Free radioactive iodine (125I) is also indicated. [Figure 9B] 1 shows single photon emission computed tomography (SPECT) and computed tomography (CT) imaging of mice with systemic AA amyloidosis 24 hours after injection of either PF.1 or 125I-hIgG1. [Figure 9C] 1 shows the biodistribution of 125I-PF.1 and 125I-hIgG1 among different tissues in AA mice 24 hours after injection. [Figure 9D] Shown are microautoradiographs of liver (left), spleen (center), and heart (right) tissues 24 hours after injection of AA mice with 125I-PF.1 or 125I-hIgG1. [Figure 10A] Immunohistochemical staining of different human tissues (heart, kidney, spleen, and brain) containing ATTR, AL, ALETC2, or Aβ amyloid with biotinylated PF.1 and Congo Red is shown. The black arrows highlight the biotinylated PF.1 bound to the tissue samples, and the white arrows highlight the presence of amyloid in the tissue samples (Congo Red staining). [Figure 10B]Immunohistochemical staining of different human tissues (heart, kidney, spleen, and brain) containing ATTR, AL, ALETC2, or Aβ amyloid with biotinylated PF.1 and Congo Red is shown. The black arrows highlight the biotinylated PF.1 bound to the tissue samples, and the white arrows highlight the presence of amyloid in the tissue samples (Congo Red staining). [Figure 10C] Immunohistochemical staining of different human tissues (heart, kidney, spleen, and brain) containing ATTR, AL, ALETC2, or Aβ amyloid with biotinylated PF.1 and Congo Red is shown. The black arrows highlight the biotinylated PF.1 bound to the tissue samples, and the white arrows highlight the presence of amyloid in the tissue samples (Congo Red staining). [Figure 10D] Immunohistochemical staining of different human tissues (heart, kidney, spleen, and brain) containing ATTR, AL, ALETC2, or Aβ amyloid with biotinylated PF.1 and Congo Red is shown. The black arrows highlight the biotinylated PF.1 bound to the tissue samples, and the white arrows highlight the presence of amyloid in the tissue samples (Congo Red staining). [Figure 10E] Immunohistochemical staining of different human tissues (heart, kidney, spleen, and brain) containing ATTR, AL, ALETC2, or Aβ amyloid with biotinylated PF.1 and Congo Red is shown. The black arrows highlight the biotinylated PF.1 bound to the tissue samples, and the white arrows highlight the presence of amyloid in the tissue samples (Congo Red staining). [Figure 11] A shows the quantified fluorescence emission from pHrodo Red in mice following injection of pHrodo Red-labeled amyloid, which was preincubated with PF.1 or alone. Increased fluorescence emission indicates amyloid phagocytosis. B shows the fluorescence emission of PF.1- and control-treated mice 12 days after injection. [Figure 12]A-D show the results of ex vivo phagocytosis assays performed with PF.1 and human IgG1 (hIgG1) control on ALκ, ALλ, ATTRv, and ATTRwt amyloid extracts. Phagocytosis was detected by labeling with the pH-sensitive dye succinimidyl-pHrodo Red fluorophore, with increased fluorescence emission indicating improved phagocytosis. ATTRwt is wild-type transthyretin-associated amyloidosis. ATTRv is variant transthyretin-associated amyloidosis. [Figure 13] A shows the results of a binding experiment testing the affinity of PF.1 for rVλWIL Aβ(1-40), ATTRwt, ATTRV, ALλ, and ALκ amyloid extracts. ATTRwt is wild-type transthyretin-associated amyloidosis. ATTRv is variant transthyretin-associated amyloidosis. B shows the results of a binding experiment testing the affinity of a human IgG1 control for the same amyloid extract panel used in FIG. 12A. [Figure 14] 1 shows the results of binding experiments testing the affinity of PF.7 for synthetic amyloid-like fibrillar α-synuclein, tau441, and Aβ(1-40). PF.7 is a VH9-D54E / VL4-N33S-VSPSV-p5R antibody-peptide fusion protein harvested after 7 days of perfusion culture. [Figure 15] 1 shows the results of an ex vivo phagocytosis assay performed on PF.1 on rVλWIL (WIL) and ALκ (TAL) fibrils in the presence or absence of 20% human serum as a source of human complement. +C indicates the presence of human serum complement. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0055] Provided herein are antibody-peptide fusion proteins that bind amyloid.

[0056] I. Definition As used herein, the singular forms "a," "an," and "the" refer to both the singular and the plural unless the context clearly indicates otherwise. The abbreviation "eg" comes from the Latin exempli gratia and is used herein to indicate a non-limiting example. Thus, the abbreviation "eg" is synonymous with the term "for example." As used herein, the term "comprises" means "including."

[0057] Ranges may be expressed herein as "about" one particular value and / or "about" another particular value. When such a range is expressed, another aspect includes one particular value in the range and / or the other particular value in the range. It is further understood that each endpoint of a range is significant 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 is understood that the particular value forms another aspect. In certain exemplary embodiments, the term "about" is understood to be within normal acceptance in the art, e.g., within two standard deviations of the mean. About may 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 the context, all numerical values ​​provided herein may be modified by the term about. Moreover, terms such as "example," "exemplary," or "illustrated" as used herein are not meant to indicate a preference, but rather to explain that the embodiment discussed thereafter is merely one example of an illustrated embodiment.

[0058] It should be further understood that all base or amino acid sizes and all molecular weight or molecular mass values ​​given for nucleic acids or polypeptides are approximate and are provided for illustration purposes. Methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present disclosure, but preferred methods and materials are described below. In case of conflict, the present specification, including explanations of terms, will control. Additionally, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0059] In order to facilitate review of the various embodiments of this disclosure, the following explanations of specific terms are provided:

[0060] The terms amyloid, amyloid deposit, amyloid fibril, and amyloid fiber refer to insoluble fibrous protein aggregates that share certain structural traits. The protein aggregates are formed, for example, by the aggregation of any of several different proteins and have a tertiary structure consisting of an ordered arrangement of beta sheets stacked perpendicular to the fiber axis. See Sunde et al., J. Mol. Biol. (1997) 273:729-39. Abnormal accumulation of amyloid in an organ can result in amyloidosis. Although they are diverse in their occurrence, all amyloids share common morphological properties in that they stain with certain dyes, e.g., Congo Red, and have a characteristic red-green birefringent appearance in polarized light after staining. Amyloids also share common ultrastructural features as well as common X-ray diffraction and infrared spectra.

[0061] Amyloidosis refers to a pathological condition or disease characterized by the presence of amyloid, e.g., the presence of amyloid deposits. "Amyloid disease" or "amyloidosis" is a disease 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 systemic AA amyloidosis, AL amyloidosis, ATTR amyloidosis, ALect2 amyloidosis, and IAPP amyloidosis in type II diabetes, are also amyloid diseases.

[0062] Amyloidogenicity refers to the ability or tendency to form amyloid deposits. For example, certain soluble monomeric proteins can undergo extensive formational changes that result in their aggregation into well-ordered, unbranched, 8-10 nm wide fibrils, which leads to the formation of amyloid aggregates. Over 30 proteins have been found to form amyloid deposits (or amyloids), for example, in humans. Not all proteins within a diverse class of proteins, such as immunoglobulin light chains, are capable of forming amyloid. That is, some proteins are nonamyloidogenic, meaning that they do not tend to form amyloid. However, other proteins in the class can form amyloid deposits and are therefore amyloidogenic. Furthermore, within the class of light chain proteins, some may be considered more "amyloidogenic" than others based on the ease with which they form amyloid fibrils. Certain light chain proteins are considered to be non-amyloidogenic or less amyloidogenic by their inability to readily form amyloid fibrils in patients or in vitro.

[0063] Animal: Living multi-cellular vertebrate organisms, a category that includes, for example, mammals and birds. The term mammal includes both human and non-human mammals. Similarly, the term "subject" includes both human and veterinary subjects. In some examples, the subject is a subject, for example, a subject suffering from an amyloid disease.

[0064] Clearance: The term "eliminate" or "clearance" refers to reducing or removing to a measurable extent. For example, the clearance of amyloid deposits described herein relates to reducing or removing deposits to a measurable or discernible extent. Clearance can result in, but is not required to result in, 100% removal. Rather, clearance can result in less than 100% removal, for example, about 10%, 20%, 30%, 40%, 50%, 60% or more removal.

[0065] Conjugate: As used herein, the term "conjugate" refers to the coupling or connection product of two or more substances, the resulting product having at least two distinct elements, e.g., at least two domains. The coupled substances can be the same or different. Such coupling can be through one or more linking groups. A "protein conjugate" results, for example, from the coupling of two or more amino acid sequences. A conjugate of two proteins results, for example, in a single protein having domains corresponding to each of the individually connected proteins.

[0066] The term "antibody" as used herein is used in the broadest sense and specifically encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired biological activity.

[0067] An "isolated" antibody is one that has been identified, separated, and / or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with research, diagnostic, or therapeutic uses of the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In some embodiments, the antibody is purified (1) until the antibody is greater than 95%, and in some embodiments greater than 99%, by weight, of the antibody, as determined, for example, by the Lowry method; (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence, for example, by use of a spinning cup sequencer; or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions, for example, using Coomassie blue or silver staining. Isolated antibodies include antibodies in situ in recombinant cells, since at least one component of the antibody's natural environment will not be present. Ordinarily, however, an isolated antibody will have been prepared by at least one purification step.

[0068] A "native antibody" is usually a heterotetrameric glycoprotein 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, but the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (VH) at one end followed by several constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end, with the constant domain of the light chain aligned with the first constant domain of the heavy chain and the variable domain of the light chain aligned with the variable domain of the heavy chain. It is believed that certain amino acid residues form an interface between the light chain variable domain and the heavy chain variable domain.

[0069] The term "constant region" refers to a portion of an immunoglobulin molecule that has a more conserved amino acid sequence than the other portion of the immunoglobulin, the variable region, which contains the antigen-binding site. The constant region contains the CR1, CR2, and CR3 domains of the heavy chain (also referred to as CH1, CH2, and CH3; collectively CH) and the CHL (or CL or CL1) domain of the light chain.

[0070] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain 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 usually the most variable parts of an antibody and contain the antigen-binding sites.

[0071] The term "variable" refers to the fact that the sequence of certain portions of the variable domains differs widely among antibodies and is used in the binding and specificity of each particular antibody to 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) in both the light and heavy chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs). Native heavy and light chain variable domains each contain four FR regions that largely adopt a beta-sheet configuration connected by three CDRs that form loops that connect and, in some cases, form part of the beta-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, together 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 binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.

[0072] The "light chains" of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two clearly distinct types, called kappa ("κ") and lambda ("λ"), based on the amino acid sequences of their constant domains.

[0073] The term IgG "isotype" or "subclass" as used herein means any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions.

[0074] Depending on the amino acid sequence of the constant domain 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, some of which can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit organization and three-dimensional configuration of the different classes of immunoglobulins are well known and are generally described, for example, in Abbas et al. Cellular and Mol. Immunology, 4th ed. (WB Saunders, Co., 2000). An antibody can 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.

[0075] The terms "full length antibody," "intact antibody," and "complete antibody" are used interchangeably herein to refer to an antibody in its substantially intact form, rather than an antibody fragment as defined below. The term particularly refers to an antibody having a heavy chain that contains an Fc region.

[0076] A "naked antibody" for purposes herein is an antibody that is not conjugated to a cytotoxic moiety or radiolabel.

[0077] An "antibody fragment" comprises a portion of an intact antibody, preferably including the antigen-binding region thereof. In some embodiments, the antibody fragments described herein are antigen-binding fragments. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0078] 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, the name of which reflects its ability to crystallize readily. Pepsin treatment produces an F(ab')2 fragment that has two antigen-binding sites and is still capable of cross-linking antigen.

[0079] An "Fv" is the minimum antibody fragment that contains a complete antigen-binding 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 chain variable domain 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 similar to that in a two-chain Fv species. In this configuration, 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 even half of an Fv containing only three CDRs specific for an antigen) has the ability to recognize and bind to an antigen, albeit with a lower affinity than the entire binding site.

[0080] Fab fragments contain the light and heavy chain variable domains and also contain the constant domain of the light chain and the first constant domain of the heavy chain (CHI.). 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 were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0081] "Single-chain Fv" or "scFv" antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Usually, 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 scFvs, see, e.g., Pluckfhun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer- Verlag, New York, 1994), pp. 269-315.

[0082] The term "monoclonal antibody" as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies that make up the population are identical except for possible mutations that may be present in minor amounts, e.g., naturally occurring mutations. Thus, the modifier "monoclonal" indicates the character of the antibody as not being a mixture of separate antibodies. In certain embodiments, such monoclonal antibodies typically include antibodies that include a polypeptide sequence that binds a target, where 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 the selected target-binding sequence can be further modified, e.g., to improve affinity to the target, humanize the target-binding sequence, improve its production in cell culture, reduce its immunogenicity in vivo, create a multispecific antibody, etc., and that antibodies that include modified target-binding sequences are also monoclonal antibodies of the 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.

[0083] The modifier "monoclonal" indicates the character of the antibody as being obtained from a population of substantially homogeneous antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in accordance with the present invention can be produced using techniques such as those described in, for example, hybridoma techniques (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, NY, 1981 )), recombinant DNA techniques (see, for example, U.S. Pat. No. 4,816,567), phage display techniques (e.g., Clackson et al, Nature, 352:624-628 (1991); Marks et al, Nature, 352:624-628 (1992)), and the like. al,J.Mol.Biol.222:581-597(1992);Sidhu et al,J.Mol.Biol.338(2):299-310(2004);Lee et al. 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 techniques for producing human or human-like antibodies in animals that have some or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits 11 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. Patent 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).

[0084] Monoclonal antibodies, as used herein, specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical 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 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 in which the antigen-binding region of the antibody is derived from, for example, an antibody generated by immunizing macaques with an antigen of interest.

[0085] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from the recipient's CDRs are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human 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. Usually, a humanized antibody will contain 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 human immunoglobulin sequences. The humanized antibody will also optionally contain 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. 1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409.

[0086] A "human antibody" is an antibody having an amino acid sequence that corresponds to that of an antibody produced by a human and / or that has been produced using any of the techniques for producing human antibodies disclosed herein. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using a variety of 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). For the preparation of human monoclonal antibodies, the methods described in Cole et al, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p.77 (1985); Boerner et al, J. Immunol., 147(1):86-95 (1991) can also be used. See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5:368-74 (2001). Human antibodies can be prepared by administering antigen to transgenic animals, e.g., immunized xenogeneic mice, that have been engineered to produce such antibodies in response to antigen challenge but whose endogenous loci have been disabled (see, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584 for XENOMOUSE™ technology). See also, e.g., Li et al, Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology.

[0087] The term "complementarity determining region" or "CDR" as used herein refers to the region of an antibody-variable domain that binds to an epitope, e.g., human amyloid fibril. Generally, an antibody contains six CDRs, three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). In native antibodies, H3 and L3 show the highest diversity of the six CDRs, and H3 in particular is believed to play a unique role in conferring superior specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, NJ, 2003)). In fact, naturally occurring camelid antibodies consisting only of heavy chains are functional and stable in the absence of light chains. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993) and Sheriff et al., Nature Struct. Biol. 3:733-736 (1996).

[0088] A number of CDR delineations are used and 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 rather to the location of structural loops (Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). In some embodiments, the CDRs may be AbM CDRs. AbM CDRs represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software. In some embodiments, the CDRs may be "contact" CDRs. "Contact" CDRs are based on analysis of available complex crystal structures. Residues from each of these CDRs are described below. [Table 1]

[0089] The CDRs may comprise "extended CDRs" as follows: in VL, 24-36 or 24-34 (L1), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3), and in VH, 26-35 (H1), 50-65 or 49-65 (preferred embodiment) (H2) and 93-102, 94-102 or 95-102 (H3). The variable domain residues are numbered according to Kabat et al. (supra) for each of these extended CDR definitions.

[0090] "Framework" or "FR" residues are those variable domain residues other than the CDR residues as herein defined.

[0091] As used herein, the term "specifically recognizes" or "specific for" refers to a measurable and reproducible interaction, such as binding, between a target and an antibody that determines 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 may be an epitope) is an antibody that binds this target with higher affinity, avidity, more readily, and / or with a longer duration than it binds to other molecules. 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 by radioimmunoassay (RIA). In certain embodiments, an antibody that specifically binds to a target has a dissociation constant (Kd) of <1M, <100 μM, <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 proteins from different species. In another embodiment, specific binding can include, but does not require, exclusive binding.

[0092] Effective amount or therapeutically effective amount: An amount of an agent sufficient to prevent, treat (including prophylactic), reduce and / or ameliorate any of the symptoms and / or underlying causes of a disorder or disease, e.g., to prevent and / or inhibit amyloidosis. In some embodiments, an "effective amount" is sufficient to reduce or eliminate symptoms of a disease. An effective amount may be administered one or more times. For example, an effective amount of a peptide is an amount sufficient to bind amyloid. The peptide may be effective when administered parenterally in an amount of more than about 1 μg / kg to about 30 mg / kg of body weight.

[0093] Inhibit: To measurably reduce. Inhibition does not require, for example, complete loss of function or complete cessation of the aspect being measured. For example, inhibiting plaque formation can mean stopping further growth of plaques, slowing down further growth of plaques, or reducing the size of plaques.

[0094] Inhibiting or Treating a Disease: Inhibiting the complete development of a disease or condition, for example, inhibiting amyloidosis. "Treatment" refers to a therapeutic intervention that improves the signs or symptoms of a disease or pathological condition after it has begun to develop. The term "improving" with respect to a disease or pathological condition refers to any observable beneficial effect of the treatment. The beneficial effect may be manifested by delayed onset of clinical symptoms of the disease in a susceptible subject, reduction in the severity of some or all clinical symptoms of the disease, slowing down the progression of the disease, improving the overall health or well-being of the subject, or by other parameters well known in the art that are specific to a particular disease. "Preventive" treatment is a treatment administered to a subject that does not show signs of the disease or does not show only early signs, with the aim of reducing the risk of developing pathology.

[0095] With respect to amyloid deposit formation, "inhibition" refers to the prevention of a reduction in the formation of amyloid deposits, for example, as compared to a control. For example, inhibition can result in about a 10%, 20%, 30%, 40%, 50%, 60% or greater reduction in amyloid deposits as compared to a control.

[0096] A label refers to any detectable compound or composition that is directly or indirectly conjugated to another molecule to facilitate its detection. Specific non-limiting examples of labels include fluorescent tags, chemiluminescent tags, haptens, enzyme conjugates, and radioisotopes. A "detectably labeled" protein, for example, means that the presence of the protein can be determined by the label associated with the protein.

[0097] Isolated: An "isolated" biological component, such as a peptide (e.g., one or more of the peptides disclosed herein), cell, nucleic acid, or serum sample, is substantially separated from, produced away from, or purified away from other biological components, such as other chromosomal and extrachromosomal DNA and RNA, and proteins, in the cells of an organism in which the component naturally occurs. "Isolated" nucleic acids, peptides, and proteins include nucleic acids and proteins purified by standard purification methods. The term also encompasses nucleic acids, peptides, and proteins prepared by recombinant expression in a cell, as well as chemically synthesized peptides and nucleic acids. The terms "isolated" or "purified" do 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 in a cell. Preferably, the preparation is purified such that the protein or peptide represents at least 50% of the total peptide or protein content of the preparation, e.g., 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.

[0098] Connect: As used herein, the terms "connect," "connected," "link," or "linked" refer 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, with or without intervening sequences or domains, e.g., in a recombinant fusion protein. Connecting also includes integrating two sequences together, e.g., placing two nucleic acid sequences together on the same nucleic acid strand such that the sequences are expressed together.

[0099] 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. Thus, the term includes nucleotide polymers in which the nucleotides and the linkages between them include synthetic analogs that do not occur in nature, such as 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, usually no more than about 50 nucleotides. When a nucleotide sequence is represented by a DNA sequence (i.e., A, T, G, C), it is understood that this also includes RNA sequences (i.e., A, U, G, C) in which "U" is replaced by "T".

[0100] Nucleotides include, but are not limited to, monomers that contain a base linked to a sugar, such as a pyrimidine, purine or synthetic analog thereof, or a base linked to an amino acid, such as peptide nucleic acid (PNA). A nucleotide is one monomer in a polynucleotide. A nucleotide sequence refers to the sequence of bases in a polynucleotide.

[0101] Conventional notations are used herein to describe nucleotide sequences: the left-hand 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 the nascent RNA transcript is referred to as the transcription direction. The DNA strand with the same sequence as the mRNA is referred to as the "coding strand"; sequences on the DNA strand with the same sequence as the mRNA transcribed from that DNA and located 5' to the 5' end of the RNA transcript are referred to as the "upstream sequence"; sequences on the DNA strand with the same sequence as the RNA and 3' to the 3' end of the coding RNA transcript are referred to as the "downstream sequence".

[0102] "cDNA" refers to a DNA that is complementary or identical to an mRNA in either single-stranded or double-stranded form.

[0103] Encoding refers to the inherent or resulting biological property of a particular sequence of nucleotides in a polynucleotide, e.g., a gene, cDNA, or mRNA, to serve as a template for the synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (e.g., rRNA, tRNA, and mRNA) or a defined sequence of amino acids. Thus, a gene encodes a protein when transcription and translation of the mRNA produced by that gene produces the protein in a cell or other biological system. Both the coding strand (whose nucleotide sequence is identical to the mRNA sequence, and is usually provided in a sequence listing), and the non-coding strand used as a template for transcription of the gene or cDNA, may 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 modified versions of each other and encode the same amino acid sequence. Protein- and RNA-encoding nucleotide sequences may include introns.

[0104] Pharmaceutically acceptable carriers: The pharma- ceutically acceptable carriers used are conventional. Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, PA, 1999. th Edition (1995) describes compositions and formulations suitable for pharmaceutical delivery of the fusion proteins disclosed herein.

[0105] The nature of the carrier will usually depend on the particular mode of administration being used. For example, parenteral formulations usually contain injectable fluids that contain pharma- ceutically and physiologically acceptable fluids as vehicles, such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol, and the like. 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, the 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, such as, for example, sodium acetate or sorbitan monolaurate.

[0106] Polypeptide: A polymer in which the monomers are amino acid residues connected to each other through amide bonds. When the amino acids are alpha-amino acids, either the L-optical isomer or the D-optical isomer may be used, with the L-isomer being preferred. The term "polypeptide" or "protein" as used herein is intended to encompass any amino acid sequence, including modified sequences, such as glycoproteins. The term "polypeptide" is specifically intended to encompass naturally occurring proteins, as well as those produced recombinantly or synthetically. In some examples, the peptide is one or more of the peptides disclosed herein.

[0107] 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 referenced protein is more pure than the protein in its native environment within a cell or production reaction chamber (as appropriate).

[0108] Recombinant: A recombinant nucleic acid is one having a sequence that is not naturally occurring or that has a sequence created by the artificial combination of two otherwise isolated segments of sequence, which can often be accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acid, e.g., by genetic engineering techniques.

[0109] 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 the percentage of identity (or similarity or homology); the higher the percentage, the more similar the two sequences are.

[0110] 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, 1988; Huang et al. Computer Appls. In the Biosciences 8, 155-65, 1992; and Pearson et al. Meth. Mol. Bio. 24, 307-31, 1994. Altschul et al. (J. Mol. Biol. 215:403-410, 1990) present detailed considerations of sequence alignment methods and homology calculations.

[0111] 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 conjunction with the sequence analysis programs blastp, blastn, blastx, tblastn, and tblastx.

[0112] Operably linked: A first nucleic acid sequence is operably linked to a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For example, a promoter is operably linked if the promoter affects the transcription or expression of a coding sequence. Usually, operably linked DNA sequences are contiguous and, where necessary to connect two protein coding regions, in the same reading frame.

[0113] Pharmaceutical agent: A chemical compound or composition capable of inducing a desired therapeutic or prophylactic effect when appropriately administered to a subject or cell.

[0114] Vector: A nucleic acid molecule that is introduced into a host cell, thereby producing a transformed host cell. A recombinant DNA vector is a vector that has recombinant DNA. A vector may contain a nucleic acid sequence that allows it to replicate in a host cell, such as an origin of replication. A vector may also contain one or more selectable marker genes and other genetic elements known in the art. A viral vector is a recombinant DNA vector that has at least some nucleic acid sequences derived from one or more viruses. The term vector includes plasmids, linear nucleic acid molecules, and adenoviral vectors and adenoviruses, as described throughout.

[0115] A subject or individual refers to a mammal, for example, a human. A subject may be a human patient. A subject may be a patient suffering from or suspected of suffering from a disease or condition, and may require treatment or diagnosis, or may require monitoring of the progression of a disease or condition. A patient may also be undergoing a treatment regimen that requires monitoring for effectiveness. In some exemplary embodiments, a subject includes an individual suffering from amyloidosis, for example, Alzheimer's disease, Huntington's or prion disease, or peripheral amyloidosis, such as that found in patients with light chain (AL) amyloidosis and type 2 diabetes.

[0116] Preferably, non-identical residue positions differ by conservative amino acid substitutions. The term "conservative amino acid substitution" refers to the interchangeability of residues with similar side chains. For example, the group of amino acids with aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; the group of amino acids with aliphatic-hydroxyl side chains is serine and threonine; the group of amino acids with amide-containing side chains is asparagine and glutamine; the group of amino acids with aromatic side chains is phenylalanine, tyrosine, and tryptophan; the group of amino acids with basic side chains is lysine, arginine, and histidine; the group of amino acids with sulfur-containing side chains is cysteine ​​and methionine. Preferred conservative amino acid substitution groups are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine.

[0117] As discussed herein, minor variations in the amino acid sequence of an antibody or immunoglobulin molecule are contemplated as being encompassed by the present invention, providing that the variations in the amino acid sequence are at least 75%, more preferably at least 80%, 90%, 95%, and most preferably 99%. In particular, conservative amino acid substitutions are contemplated. Conservative substitutions are those that occur within a family of amino acids that are related in their side chains. Genetically encoded amino acids are usually divided into the following families: (1) acidic amino acids are aspartic acid, glutamic acid; (2) basic amino acids are lysine, arginine, histidine; (3) nonpolar 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. Hydrophilic amino acids include arginine, asparagine, aspartic acid, glutamine, glutamic acid, histidine, lysine, serine, and threonine. Hydrophobic amino acids include alanine, cysteine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, tyrosine, and valine. Other families of amino acids include (i) the aliphatic hydroxy family, serine and threonine; (ii) the amide-containing family, asparagine and glutamine; (iii) the aliphatic family, alanine, valine, leucine, and isoleucine; and (iv) the aromatic family, phenylalanine, tryptophan, and tyrosine. For example, it is reasonable to predict that the independent replacement of leucine with isoleucine or valine, aspartic acid with glutamic acid, threonine with serine, or the 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 amino acid in a framework region. Whether an amino acid change results in a functional peptide can be determined by assaying the specific activity of the polypeptide derivative, the assays being described in detail herein.Fragments or analogs of antibody or immunoglobulin molecules can be readily prepared by one of skill in the art. Preferred amino and carboxy termini of the fragments or analogs occur near boundaries of functional domains. Structural and functional domains can be identified by comparison of 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 for identifying protein sequences that fold into known three-dimensional structures are known. (Bowie et al. Science 253:164 (1991). Thus, the foregoing examples demonstrate that one of skill in the art can recognize sequence motifs and structural conformations that can be used to define structural and functional domains in accordance with the present invention.

[0118] Preferred amino acid substitutions are those that (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) modify binding affinity for forming protein complexes, (4) modify binding affinity, and (4) confer or modify other physicochemical or functional properties of such analogs. Analogs may include various mutant proteins of sequences other than naturally occurring peptide sequences. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) may be made in naturally occurring sequences (preferably in a portion of a polypeptide outside the domain(s) that form intermolecular contacts). Conservative amino acid substitutions should not substantially change the structural characteristics of the parent sequence (e.g., the replacement amino acid should not tend to break the helix that occurs in the parent sequence or destroy other types of secondary structures that characterize 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, NY (1991)); and Thornton et al. Nature 354:105 (1991).

[0119] Except for CDR1 in VH, CDRs usually contain amino acid residues that form hypervariable loops. CDRs also contain "specificity determining residues" or "SDRs", which are residues that contact the antigen. SDRs are contained within regions of CDRs called truncated 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 residues 31-34 of LI, 50-55 of L2, 89-96 of L3, 31-35B of HI, 50-58 of H2, and 95-102 of H3. (See Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)).

[0120] II. Antibody-Peptide Fusion Proteins Provided herein are antibody-peptide fusion proteins that target amyloid. Such antibody-peptide fusion proteins include, for example, an antibody, a fragment thereof, an amyloid-reactive peptide linked to an antibody by an N-terminal or C-terminal light chain protein extension of the antigen-binding (Fab) region, or via the N-terminus or C-terminus of the antibody's heavy chain, thereby forming a peptide-antibody fusion, where the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. The antibody-peptide fusion proteins can be used to treat a subject having or suspected of having amyloidosis, for example, by administering the antibody-peptide fusion protein to the subject.

[0121] In some embodiments, provided herein are antibody-peptide fusion proteins comprising an amyloid-reactive 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 an 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 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 some embodiments, the amyloid-reactive peptide and the antibody are linked at the N- and / or C-terminus of the light chain and / or the N- and / or C-terminus of the heavy chain. In some embodiments, the antibody-peptide fusion protein comprises multiple amyloid-reactive peptides linked to an antibody. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a peptide spacer. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the heavy chain. In some embodiments, the antibody is a full-length antibody. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence shown in Table 1 below.

[0122] In some embodiments, the amyloid reactive peptide antibody fusion comprises a heavy chain comprising, in order from N to C terminal, an amyloid reactive peptide, a spacer, a VH, a CH1, a CH2, and a CH3. In some embodiments, the amyloid reactive peptide antibody fusion comprises a heavy chain comprising, in order from N to C terminal, a VH, a CH1, a CH2, a CH3, a spacer, and an amyloid reactive peptide. In some embodiments, the amyloid reactive peptide antibody fusion comprises a light chain comprising, in order from N to C terminal, an amyloid reactive peptide, a spacer, a VL, and a CL. In some embodiments, the amyloid reactive peptide antibody fusion comprises a light chain comprising, in order from N to C terminal, a VL, and a CL, a spacer, and an amyloid reactive peptide. [Table 2]

[0123] Without being bound to any particular theory, it is believed that the amyloid reactive peptide of the antibody-peptide fusion protein targets the antibody-peptide fusion protein to amyloid deposits when administered to a subject. The Fc domain then induces an immune response at the site of the amyloid, thereby resulting in the clearance of the amyloid, for example, by opsonization. It is also believed that the antibody-peptide fusion protein has a longer half-life than the amyloid reactive peptide alone. For example, the circulating half-life of IgG in humans is approximately 21 days, while the half-life of the amyloid reactive peptide alone in humans is approximately 11 hours. Thus, the Ig extends the half-life of the antibody-peptide fusion protein in circulation. In some embodiments, the half-life of the antibody-peptide fusion protein is increased by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or more compared to the amyloid reactive peptide alone. In this way, the antibody-peptide fusion protein, when administered to a subject, is able to exert its immunostimulatory effect for a longer period at the site of the amyloid deposit, thereby increasing the immune response at the site of the amyloid deposit.

[0124] In some embodiments, the amyloid reactive peptide of the antibody-peptide fusion protein described herein comprises an amino acid sequence that is at least 80%, 85%, 90% or more identical to an amino acid sequence set forth as any one of SEQ ID NOs: 1-13, e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to an amino acid sequence set forth as any one of SEQ ID NOs: 1-13. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer that comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer that comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid reactive peptide linked to the antibody or functional fragment thereof may comprise or consist of about 10 to about 55 amino acids. The amyloid-reactive peptides of the invention can comprise or consist of, for example, 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. Such peptides are described, for example, in International Patent Application WO2016032949 and Wall et al. (PLoS One. 2013 Jun 4;8(6):e66181), which are incorporated herein in their entireties. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or more sequence identity to any one of the amino acid sequences set forth as SEQ ID NOs: 1-13. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or more sequence identity to any one of the amino acid sequences set forth as SEQ ID NOs: 1-13.In some embodiments, the amyloid reactive peptide comprises an amino acid sequence set forth as SEQ ID NO: 1-13, including one or more amino acid substitutions. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence set forth in SEQ ID NO: 1. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence set forth in SEQ ID NO: 13. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence set forth in any one of SEQ ID NOs: 1-13.

[0125] The amino acids forming all or part of the amyloid-reactive peptide bound to the antibody or fragment thereof can 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, etc. The amino acids forming the peptides of the invention can be one or more of the 20 common amino acids found in naturally occurring proteins, or one or more modified and unconventional amino acids. Antibody-peptide fusion proteins can be made by any technique known to those of skill in the art, including chemical synthesis or recombinant means using standard molecular biology techniques.

[0126] In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the antibody comprises one, two, three, four, five, or six CDRs of an antibody shown in Table 2. [Table 3]

[0127] In certain embodiments, an antibody-peptide fusion protein comprises an antibody, the antibody comprising a VH comprising (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 linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0128] In certain embodiments, the antibody-peptide fusion protein comprises an antibody, the antibody comprising a VL comprising (a) a CDR-Ll 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 linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86.

[0129] In one embodiment, the antibody-peptide fusion protein comprises an antibody comprising a VL comprising the amino acid sequence of SEQ ID NO: 16 and a VH comprising the amino acid sequence of SEQ ID NO: 15, wherein the antibody is linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. The amino acid sequences of SEQ ID NO: 15 and SEQ ID NO: 16 are provided below. m11-1F4 VH SEQ ID NO: 15 QVQLKESGPGLVAPSQSLSITCTVSGFSLSSYGVSWVRQPPGKGLEWLGVIWGDGSTNYHPNLMSRLSISKDISKSQVLFKLNSLQTDDTATYYCVTLDYWGQGTSVTVSS m11-1F4 VL SEQ ID NO: 16 DVVMTQTPLSLPVSLGDQASISCRSSQSLVHRNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGLYFCFQTTYVPNTFGGGTKLEIK

[0130] In another aspect, an antibody-peptide fusion protein comprises an antibody, the antibody comprising a VH comprising a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 17, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 18, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 19, and a VL comprising a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 21, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 22, the antibody being linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising any of the amino acid sequences listed in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide comprising the amino acid sequence of any one of SEQ ID NOs: 1-13.

[0131] In another aspect, an antibody-peptide fusion protein comprises an antibody, the antibody comprising a VH CDR1, a VH CDR2, and a VH CDR3 of a VH having the sequence set forth in SEQ ID NO: 15 and a VL CDR1, a VL CDR2, and a VL of a VL having the sequence set forth in SEQ ID NO: 16, the antibody being linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising any of the amino acid sequences listed in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide comprising the amino acid sequence of any of SEQ ID NOs:1-13.

[0132] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15 and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15 without a C-terminal lysine residue, and a VL comprising the amino acid sequence of SEQ ID NO: 16, wherein the antibody is linked to an amyloid reactive peptide.

[0133] In another aspect, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising any of the amino acid sequences listed in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide comprising any of the amino acid sequences of SEQ ID NO: 1-13. In some embodiments, the amyloid reactive peptide is linked to the N-terminus or C-terminus of the antibody light chain or the N-terminus or C-terminus of the heavy chain. In some embodiments, the antibody also comprises a spacer amino acid sequence between the amyloid reactive peptide and the N-terminus or C-terminus of the antibody light chain or the N-terminus or C-terminus of the heavy chain. In some embodiments, the spacer is a peptide spacer. In some embodiments, the spacer is flexible or rigid. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 83 and 86. In some embodiments, the spacer comprises the amino acid sequence shown in SEQ ID NO: 83. In some embodiments, the amyloid-reactive peptide is linked to the C-terminus of the light chain. In some embodiments, the amyloid-reactive peptide shown in SEQ ID NO: 2 is linked to the C-terminus of the light chain via a spacer comprising the amino acid sequence shown in SEQ ID NO: 83.

[0134] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15, wherein the heavy chain is linked to an amyloid-reactive peptide comprising any of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15, wherein the heavy chain is linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15, wherein the heavy chain is linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15, wherein the heavy chain is linked to an amyloid-reactive peptide comprising any of the amino acid sequences of SEQ ID NO: 1-13. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86.

[0135] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, the light chain being linked to an amyloid-reactive peptide comprising any of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, the light chain being linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, the light chain being linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, the light chain being linked to an amyloid-reactive peptide comprising any of the amino acid sequences of SEQ ID NO: 1-13. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86.

[0136] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15 and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to an amyloid-reactive peptide comprising any of the amino acid sequences in Table 1. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0137] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15 and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the amyloid reactive peptide is linked to the light chain at the N-terminus. In some embodiments, the amyloid reactive peptide is linked to the light chain at the C-terminus. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0138] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15 and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to an amyloid reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the amyloid reactive peptide is linked to the light chain at the N-terminus. In some embodiments, the amyloid reactive peptide is linked to the light chain at the C-terminus. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0139] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain comprising a VH comprising the amino acid sequence of SEQ ID NO: 15 and a light chain comprising a VL comprising the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to an amyloid-reactive peptide comprising the amino acid sequence of any of SEQ ID NOs: 1-13. In some embodiments, the amyloid-reactive peptide is linked to the light chain at the N-terminus. In some embodiments, the amyloid-reactive peptide is linked to the light chain at the C-terminus. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0140] Also provided herein are antibody-peptide fusion proteins comprising a humanized antibody that binds to human amyloid fibrils fused to an amyloid-reactive peptide. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody described herein. In some embodiments, the humanized antibody comprises a humanized VH and / or VL sequence derived from m11-1F4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody described in International Application No. PCT / US2020 / 060596, which is incorporated herein by reference in its entirety. Exemplary amino acid sequences of the humanized VH and VL regions are provided below in Tables 3-4. In Tables 3-4, the CDR sequences are underlined, and the backmutated residues as well as the additional mutations introduced into the humanized variants VL4 and VH9 are in bold and italicized. The additional mutations introduced into VL4 and VH9 are listed in the IgG column of Tables 3-4; these mutations are numbered relative to the N-terminus of the VL or VH. The CDR amino acid sequences for variants of VL4 and VH9 with modified CDRs are presented in Tables 5 and 6 relative to VL4 and VH9 below. [Table 4] [Table 5-1] [Table 5-2] [Table 6] [Table 7]

[0141] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the humanized antibody comprises one, two, three, four, five, or six CDRs of an antibody set forth in Table 2. In some embodiments, the humanized antibody comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, of a VH having the sequence set forth in SEQ ID NO: 15, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, of a VL having the sequence set forth in SEQ ID NO: 16. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0142] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20 with one or more conservative amino acid substitutions, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21 with one or more conservative amino acid substitutions, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22 with one or more conservative amino acid substitutions, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17 with one or more conservative amino acid substitutions, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18 with one or more conservative amino acid substitutions, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19 with one or more conservative amino acid substitutions. In some embodiments, the humanized antibody comprises one, two, three, four, five, or six CDRs of an antibody set forth in Table 2 with one or more conservative amino acid substitutions. In some embodiments, the humanized antibody comprises CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequences of CDR-H1, CDR-H2, and CDR-H3, respectively, of a VH having a sequence set forth in SEQ ID NO: 15 with one or more conservative amino acid substitutions, and CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequences of CDR-L1, CDR-L2, and CDR-L3, respectively, of a VL having a sequence set forth in SEQ ID NO: 16 with one or more conservative amino acid substitutions. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0143] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: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 a VH comprising 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 antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising 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 a VH comprising 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:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0144] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 71, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0145] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 72, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0146] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 73, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0147] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 74, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0148] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 75, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0149] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 76, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0150] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 77, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0151] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 78, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0152] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 79, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0153] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 80, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0154] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 64-70, a CDR-L2 comprising an amino acid sequence set forth in SEQ ID NO: 21, and a CDR-L3 comprising an amino acid sequence set forth in SEQ ID NO: 22, and a VH comprising a CDR-H1 comprising an amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence set forth in SEQ ID NO: 81, and a CDR-H3 comprising an amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86.

[0155] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:83-86.

[0156] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:65, 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:83-86.

[0157] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:66, 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:83-86.

[0158] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:67, 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:83-86.

[0159] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:68, 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:83-86.

[0160] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:69, 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO:71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO:83-86.

[0161] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 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 a VH comprising a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 17, a CDR-H2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 71-81, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 19. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 83-86.

[0162] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising 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 a VH comprising 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. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, and 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83 and ... the amino acid sequence set forth in SEQ ID NO:83. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence set forth in SEQ ID NO:2. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to the amyloid reactive peptide set forth in SEQ ID NO:2 via a spacer comprising the amino acid sequence set forth in SEQ ID NO:83. In some embodiments, the antibody-peptide fusion protein is a full-length antibody. In some embodiments, the antibody-peptide fusion protein has an IgG1 isotype. In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, in order from N to C-terminus, a VL, a CL, a spacer, and an amyloid reactive peptide. In some embodiments, the amyloid reactive peptide is fused to the C-terminus of the light chain via a spacer.

[0163] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising the amino acid sequence of a VL shown in Table 3. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL selected from the group consisting of VL2, VL3, VL4, VL4-N33S, VL4-N33Q, VL4-N33E, VL4-N33A, VL4-N33H, VL4-G34A, or VL4-G34V shown in Table 3. In some embodiments, the VL comprises the amino acid sequence shown in the group consisting of SEQ ID NOs: 32-42. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0164] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising the amino acid sequence of a VH as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, wherein the humanized antibody comprises a VH selected from the group consisting of VH2, VH3, VH4, VH5, VH6, VH7, VH8, VH9, VH10, VH9-D54S, VH9-D54Q, VH9-D54E, VH9-D54A, VH9-D54H, VH9-G55A, VH9-G55V, VH9-M64V, VH9-M64I, VH9-M64L, or VH9-M64A, as set forth in Table 4. In some embodiments, the VH comprises the amino acid sequence as set forth in the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0165] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 32, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0166] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 33, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0167] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 34, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0168] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 35, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0169] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 36, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0170] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:37, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0171] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:38, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0172] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:39, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0173] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:40, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0174] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:41, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0175] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 42, and a VH comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0176] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0177] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 44. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0178] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 45. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0179] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 46. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0180] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 47. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0181] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 48. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0182] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 49. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0183] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 50. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0184] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 51. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0185] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 52. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0186] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 53. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0187] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 54. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0188] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 55. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0189] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 56. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0190] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 57. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0191] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 58. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0192] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 59. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0193] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 60. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0194] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 61. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0195] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 62. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0196] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-42, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 63. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0197] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 34, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 48. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0198] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 35, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 51. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0199] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:35, and a VH comprising the amino acid sequence set forth in SEQ ID NO:52. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0200] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:35, and a VH comprising the amino acid sequence set forth in SEQ ID NO:50. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0201] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO: 35, and a VH comprising the amino acid sequence set forth in SEQ ID NO: 49. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0202] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:36, and a VH comprising the amino acid sequence set forth in SEQ ID NO:55. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer. In some embodiments, the amyloid-reactive peptide is fused to the C-terminus of the light chain via a spacer. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83 and 86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising the amino acid sequence set forth in SEQ ID NO:83. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to the amyloid reactive peptide set forth in SEQ ID NO: 2 via a spacer comprising the amino acid sequence set forth in SEQ ID NO: 83.

[0203] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL of VL4 as set forth in Table 3, and a VH of VH9 as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence as set forth in SEQ ID NO:35, and a VH comprising the amino acid sequence as set forth in SEQ ID NO:51. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0204] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL of VL4-N33S as set forth in Table 3, and a VH of VH9-D54E as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence set forth in SEQ ID NO:36, and a VH comprising the amino acid sequence set forth in SEQ ID NO:55. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0205] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL of VL3 as set forth in Table 3 and a VH of VH6 as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence as set forth in SEQ ID NO:34 and a VH comprising the amino acid sequence as set forth in SEQ ID NO:48. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, 83-86. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86. In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:1-13.

[0206] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL of VL4 as set forth in Table 3 and a VH of VH10 as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence as set forth in SEQ ID NO:35 and a VH comprising the amino acid sequence as set forth in SEQ ID NO:52. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL of VL4 as set forth in Table 3 and a VH of VH8 as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence as set forth in SEQ ID NO:35 and a VH comprising the amino acid sequence as set forth in SEQ ID NO:50. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL of VL4 as set forth in Table 3 and a VH of VH7 as set forth in Table 4. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a VL comprising the amino acid sequence as set forth in SEQ ID NO:35 and a VH comprising the amino acid sequence as set forth in SEQ ID NO:69. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13.

[0207] In some embodiments, an antibody-peptide fusion protein comprising a humanized antibody comprises an amyloid reactive peptide. In some embodiments, the amyloid reactive peptide comprises one or more of the peptides set forth in Table 1. In certain embodiments, the amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13. 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: 2.

[0208] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprises a light chain. In some embodiments, the amyloid reactive peptide is fused to the N-terminus of the light chain. In some embodiments, the amyloid reactive peptide is fused to the C-terminus of the light chain. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprises a light chain, and the amyloid reactive peptide is fused to the N-terminus of the light chain by a spacer. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprises a light chain, and the amyloid reactive peptide is fused to the C-terminus of the light chain by a spacer. In some embodiments, the spacer is a peptide spacer. In some embodiments, the spacer is a soft spacer. In some embodiments, the spacer comprises glycine and serine residues. In some embodiments, the spacer comprises the amino acid sequence GGGYS. In some embodiments, the spacer comprises the amino acid sequence set forth in SEQ ID NO:27. In some embodiments, the spacer is a rigid spacer. In some embodiments, the spacer is uncharged. In some embodiments, the spacer comprises the amino acid sequence set forth in SEQ ID NOs:83-86.

[0209] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprises a light chain. In some embodiments, the amyloid reactive peptide is fused to the N-terminus of the heavy chain. In some embodiments, the amyloid reactive peptide is fused to the C-terminus of the heavy chain. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprises a heavy chain, and the amyloid reactive peptide is fused to the N-terminus of the heavy chain by a spacer. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprises a heavy chain, and the amyloid reactive peptide is fused to the C-terminus of the heavy chain by a spacer. In some embodiments, the spacer is a peptide spacer. In some embodiments, the spacer is a soft spacer. In some embodiments, the spacer is a rigid spacer. In some embodiments, the spacer is uncharged. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:83-86.

[0210] In certain embodiments, the antibody-peptide fusion protein may include a spacer sequence of amino acids between the C- or N-terminus of the light chain or the C- or N-terminus of the heavy chain and the amyloid-reactive peptide. In certain embodiments, the peptide-Ig conjugate may include a spacer sequence of amino acids between the N-terminus of the peptide and a leader sequence required for secretion of the Ig-peptide from cells expressing the reagent. In some embodiments, the spacer is a soft spacer. In some embodiments, the spacer is a hard spacer peptide. In some embodiments, the spacer is uncharged. In some embodiments, the spacer peptide may include or consist of about 3 to about 55 amino acids. The spacer peptide 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 adjacent to each other in sequence. For example, two nucleic acid sequences may be adjacent to each other as described herein, but still include an intervening spacer sequence. In some embodiments, the spacer peptide comprises the amino acid sequence shown in Table 9 below. In some embodiments, the spacer comprises the amino acid sequence set forth in SEQ ID NOs:83-86. [Table 8]

[0211] In some embodiments, one or more of the peptides shown in Table 1 may be linked to a humanized antibody or functional fragment thereof via the C- or N-terminus of the light chain protein or the C- or N-terminus of the heavy chain, thereby forming an antibody-peptide fusion protein comprising a humanized antibody. That is, any of the sequences identified in Table 1 below may be linked independently or simultaneously to the heavy or light chain of a humanized antibody or functional fragment thereof to form an antibody-peptide fusion protein. For example, two of the amyloid reactive peptides may be linked to a single antibody, for example, by linking an amyloid reactive peptide amino acid sequence to the N-terminus of the humanized antibody light chain, or by connecting an amyloid reactive peptide amino acid sequence to the C-terminus of the humanized antibody light chain.

[0212] In some embodiments, the antibody-peptide fusion protein comprises a light chain that further comprises a light chain constant region (e.g., comprising CL1) and comprises a heavy chain that comprises a heavy chain constant region (e.g., comprising CH1, CH2, and CH3). In some embodiments, the antibody-peptide fusion protein comprises two light chains and two heavy chains.

[0213] In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, from N- to C-terminus, an amyloid reactive peptide and a light chain. In some embodiments, the light chain comprises, from N- to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N- to C-terminus, a VH, a CH1, a CH2, and a CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, first and second light chains comprising, from N- to C-terminus, an amyloid reactive peptide, a spacer, a variable light chain region, and a constant light chain region, and first and second heavy chains comprising, from N- to C-terminus, a VH, a CH1, a CH2, and a CH3, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0214] In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a light chain and a light chain comprising an amyloid-reactive peptide. In some embodiments, the light chain comprises, from N- to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a heavy chain comprising, from VH, CH1, CH2, and CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, first and second light chains comprising, from N- to C-terminus, a variable light chain region, a constant light chain region, a spacer, and an amyloid-reactive peptide, and first and second heavy chains comprising, from N- to C-terminus, a VH, CH1, CH2, and CH3, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0215] In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a light chain comprising an amyloid reactive peptide, a spacer peptide, and a light chain. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 83-86. In some embodiments, the light chain comprises, from N- to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a heavy chain comprising a VH, a CH1, a CH2, and a CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises first and second light chains comprising, from N to C-terminus, an amyloid reactive peptide, a spacer, VL, and CL1, and first and second heavy chains comprising, from N to C-terminus, VH, CH1, CH2, and CH3, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0216] In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a light chain, a spacer peptide, and a light chain comprising an amyloid-reactive peptide. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 83-86. In some embodiments, the light chain comprises, from N- to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a heavy chain comprising a VH, a CH1, a CH2, and a CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises first and second light chains comprising, from N to C-terminus, a VL, a CL1, a spacer, and an amyloid-reactive peptide, and first and second heavy chains comprising, from N to C-terminus, a VH, a CH1, a CH2, and a CH3, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0217] In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a secretory leader peptide, a first spacer peptide, an amyloid-reactive peptide, a second spacer peptide, and a light chain. In some embodiments, the first spacer peptide comprises the amino acid sequence of SEQ ID NO:23. In some embodiments, the first spacer peptide comprises the amino acid sequence of SEQ ID NO:27. In some embodiments, the second spacer peptide comprises the amino acid sequence of SEQ ID NO:24. In some embodiments, the first and / or second spacer peptide comprises the amino acid sequence of SEQ ID NO:83-86. In some embodiments, the light chain comprises, from N- to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a heavy chain comprising a VH, a CH1, a CH2, and a CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the VH is any one of the VHs described herein.

[0218] In some embodiments, the antibody-peptide fusion protein comprises, from N to C terminus, a secretory leader peptide, a first spacer peptide, a light chain, a second spacer peptide, and an amyloid-reactive peptide. In some embodiments, the first spacer peptide comprises the amino acid sequence of SEQ ID NO:23. In some embodiments, the first spacer peptide comprises the amino acid sequence of SEQ ID NO:27. In some embodiments, the second spacer peptide comprises the amino acid sequence of SEQ ID NO:24. In some embodiments, the first and / or second spacer peptide comprises the amino acid sequence of SEQ ID NO:83-86. In some embodiments, the light chain comprises, from N to C terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises, from N to C terminus, a heavy chain comprising a VH, a CH1, a CH2, and a CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the VH is any one of the VHs described herein.

[0219] In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, from N to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N to C-terminus, an amyloid reactive peptide, a VH, a CH1, a CH2, and a CH3. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises first and second light chains comprising, from N to C-terminus, a VL, and a CL1, and first and second heavy chains comprising, from N to C-terminus, an amyloid reactive peptide, a VH, a CH1, a CH2, and a CH3, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0220] In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, from N to C-terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N to C-terminus, a VH, a CH1, a CH2, a CH3, and an amyloid-reactive peptide. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises first and second light chains comprising, from N to C-terminus, a VL, and a CL1, and first and second heavy chains comprising, from N to C-terminus, a VH, a CH1, a CH2, a CH3, and an amyloid-reactive peptide, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0221] In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, from N to C terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N to C terminus, an amyloid reactive peptide, a spacer peptide, a VH, a CH1, a CH2, and a CH3. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 83-86. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises first and second light chains comprising, from N to C terminus, a VL, and a CL1, and first and second heavy chains comprising, from N to C terminus, an amyloid reactive peptide, a spacer, a VH, a CH1, a CH2, and a CH3, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0222] In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, from N to C terminus, a VL and a CL1. In some embodiments, the VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N to C terminus, a VH, a CH1, a CH2, a CH3, a spacer peptide, and an amyloid-reactive peptide. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the spacer peptide comprises the amino acid sequence of SEQ ID NO: 83-86. In some embodiments, the VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises first and second light chains comprising, from N to C terminus, a VL, and a CL1, and first and second heavy chains comprising, from N to C terminus, a VH, a CH1, a CH2, a CH3, a spacer, and an amyloid-reactive peptide, wherein the CH2 and CH3 of the first and second heavy chains form a dimer.

[0223] In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising, from N- to C-terminus, an amyloid reactive peptide, a spacer peptide, and a light chain. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence of SEQ ID NO:2. In some embodiments, the spacer comprises the amino acid sequence of SEQ ID NO:83. In some embodiments, the light chain comprises a VL comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N- to C-terminus, a VH, a CH1, a CH2, and a CH3. In some embodiments, the heavy chain comprises a VH comprising the amino acid sequence of SEQ ID NO:55. In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising the amino acids set forth in SEQ ID NO:87, and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the antibody-peptide fusion protein comprises a first polypeptide and a second polypeptide comprising an amyloid reactive peptide linked to the N-terminus of a light chain of an antibody that binds to human amyloid fibrils, and a third and a fourth polypeptide comprising a heavy chain of an antibody that binds to human amyloid fibrils, wherein the first and second polypeptides comprise the amino acid sequence set forth in SEQ ID NO: 87, and the third and fourth polypeptides comprise the amino acid sequence set forth in SEQ ID NO: 91. In some embodiments, the antibody-peptide fusion protein comprises the structure shown in FIG.

[0224] In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a heavy chain, a spacer peptide, and a heavy chain comprising an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence of SEQ ID NO:2. In some embodiments, the spacer comprises the amino acid sequence of SEQ ID NO:83. In some embodiments, the light chain comprises a VL comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, the heavy chain comprises a VH comprising the amino acid sequence of SEQ ID NO:55. In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:88, and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:92. In some embodiments, the antibody-peptide fusion protein comprises a first polypeptide and a second polypeptide comprising a light chain of an antibody that binds to human amyloid fibrils, and a third and a fourth polypeptide comprising an amyloid-reactive peptide linked to the C-terminus of the heavy chain of an antibody that binds to human amyloid fibrils, wherein the first polypeptide and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:88, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:92. In some embodiments, the antibody-peptide fusion protein comprises the structure shown in FIG.

[0225] In some embodiments, the antibody peptide fusion comprises a light chain comprising, from N-terminus to C-terminus, a variable light chain region, a constant light chain region, a spacer, and an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence of SEQ ID NO:2. In some embodiments, the spacer comprises the amino acid sequence of SEQ ID NO:83. In some embodiments, the light chain comprises a VL comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, the heavy chain comprises a VH comprising the amino acid sequence of SEQ ID NO:55. In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:89, and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the antibody-peptide fusion protein comprises a first polypeptide and a second polypeptide comprising an amyloid-reactive peptide linked to the C-terminus of a light chain of an antibody that binds human amyloid fibrils, and a third and a fourth polypeptide comprising a heavy chain of an antibody that binds human amyloid fibrils, wherein the first polypeptide and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:89, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the antibody-peptide fusion protein comprises the structure shown in FIG.

[0226] In some embodiments, the antibody-peptide fusion protein comprises, from N- to C-terminus, a light chain, a spacer peptide, and a light chain comprising an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence of SEQ ID NO:2. In some embodiments, the spacer comprises the amino acid sequence of SEQ ID NO:86. In some embodiments, the light chain comprises a VL comprising the amino acid sequence of SEQ ID NO:36. In some embodiments, the heavy chain comprises a VH comprising the amino acid sequence of SEQ ID NO:55. In some embodiments, the antibody-peptide fusion protein comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:90, and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the antibody-peptide fusion protein comprises a first polypeptide and a second polypeptide comprising an amyloid-reactive peptide linked to the C-terminus of a light chain of an antibody that binds human amyloid fibrils, and a third and a fourth polypeptide comprising a heavy chain of an antibody that binds human amyloid fibrils, wherein the first polypeptide and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:90, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the antibody-peptide fusion protein comprises the structure shown in FIG.

[0227] In some embodiments, the antibody-peptide fusion protein comprises an antibody that binds to amyloid fibrils, the antibody comprising a first and a second polypeptide, each comprising a light chain of the antibody, and a third and a fourth polypeptide, each comprising a heavy chain of the antibody. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide linked to the N-terminus or C-terminus of the light or heavy chain. In some embodiments, the first and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:87, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the first and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:88, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:92. In some embodiments, the first and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:89, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the first and the second polypeptide comprise the amino acid sequence set forth in SEQ ID NO:90, and the third and the fourth polypeptide comprise the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the amyloid reactive peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-13. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the amyloid reactive peptide comprises the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid reactive peptide set forth in SEQ ID NO: 2 via a spacer comprising the amino acid sequence set forth in SEQ ID NO: 83.

[0228] In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide comprising the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. In some embodiments, the antibody-peptide fusion protein comprises an antibody that binds to human amyloid fibrils. In some embodiments, the antibody comprises a variable heavy chain (VH) and a variable light chain (VL), where 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, and 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. In some embodiments, the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer. In some embodiments, the antibody-peptide fusion comprises a light chain comprising, from N-terminus to C-terminus, a variable light chain region, a constant light chain region, a spacer, and an amyloid-reactive peptide. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the spacer comprises the amino acid sequence shown in SEQ ID NO: 83.

[0229] In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid deposits or fibrils. In some embodiments, the antibody-peptide fusion proteins bind to one or more amyloidogenic peptides in amyloid. In some embodiments, the amyloid to which the antibody-peptide fusion protein binds comprises an amyloidogenic λ6 variable domain protein (Vλ6Wil) or an amyloidogenic immunoglobulin light chain (AL), Aβ(1-40) amyloid-like fibril or amyloidogenic Aβ precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid to which the antibody-peptide fusion protein binds includes amyloidogenic forms of immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTR), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), gelsolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or other amyloidogenic peptides such as IAAP, ALκ4, ALλ1, etc. The amyloidogenic peptide to which the antibody-peptide fusion protein binds 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, amyloid is part of the pathology of a disease.

[0230] In some embodiments, the antibodies provided herein specifically bind to amyloid light chain fibrils. In some embodiments, the amyloid-reactive peptides bind to various amyloid fibrils, such as amyloidogenic λ6 variable domain protein (Vλ6Wil) or amyloidogenic immunoglobulin light chain (AL), Aβ(1-40) amyloid-like fibrils or amyloidogenic Aβ precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid to which the antibody-peptide fusion protein binds includes amyloidogenic forms of immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTR), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), gelsolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP); α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or other amyloidogenic peptides such as IAAP, ALκ4, ALλ1, etc. In some embodiments, the amyloid reactive peptide binds to heparan sulfate glycosaminoglycans.

[0231] In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid deposits or fibrils. In some embodiments, the amyloid deposits or fibrils are located in one or more organs. In some embodiments, the amyloid deposits are located in one or more tissue types. 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 blood. In some embodiments, the antibody-peptide fusion proteins bind to amyloid deposits or 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 to amyloid deposits or fibrils located in the liver, spleen, heart, kidney, brain, muscle, pancreas, stomach, upper intestine, lower intestine, and / or blood.

[0232] In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide comprising the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. In some embodiments, the antibody-peptide fusion comprises, from N-terminal to C-terminal, a light chain comprising a variable light region, a constant light region, a spacer, and an amyloid-reactive peptide. In some embodiments, the antibody-peptide fusion protein comprises an antibody that binds to human amyloid fibrils. In some embodiments, the antibody comprises a variable heavy chain (VH) and a variable light chain (VL), wherein 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, and 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. In some embodiments, the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, 83-86. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the spacer comprises the amino acid sequence set forth in SEQ ID NO: 83. In some embodiments, the antibody-peptide fusion protein exhibits pan-amyloid reactivity. In some embodiments, the antibody-peptide fusion protein exhibits reactivity to amyloid deposits or fibrils located in the liver, spleen, heart, kidney, brain, muscle, pancreas, stomach, upper intestine, lower intestine, and / or blood.

[0233] In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid deposits or fibrils with high binding affinity. In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid substrates with high binding affinity. In some embodiments, the binding affinity is less than 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 5 nM, 4 nM, 3 nM, 2 nM, or 1.5 nM. In some embodiments, the binding affinity is less than 500 nM. In some embodiments, the binding affinity is less than 100 nM. In some embodiments, the binding affinity is less than 10 nM. In some embodiments, the binding affinity is less than 1.5 nM. In some embodiments, the binding affinity is 0.05 nM to 100 nM, 0.1 nM to 50 nM, 0.2 nM to 25 nM, 0.3 nM to 10 nM, 0.4 nM to 5 nM, 0.5 nM to 2 nM, 0.6 nM to 1 nM, or 0.2 nM to 1.5 nM. In some embodiments, the binding affinity is the same or different for different amyloid substrates. In some embodiments, the binding affinity is the same or different for human amyloid substrates. In some embodiments, the binding affinity is the same or different for synthetic amyloid substrates.

[0234] In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide comprising the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. In some embodiments, the antibody-peptide fusion protein comprises an antibody that binds to human amyloid fibrils. In some embodiments, the antibody comprises a variable heavy chain (VH) and a variable light chain (VL), wherein 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, and 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. In some embodiments, the amyloid-reactive peptide and the antibody are linked at the C-terminus of the light chain. In some embodiments, the amyloid-reactive peptide is linked to the antibody via a spacer. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:23-24, 27, and 83-86. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 83-86. In some embodiments, the spacer comprises an amino acid sequence as set forth in SEQ ID NO: 83. In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid deposits or fibrils with a binding affinity represented by an EC50 binding affinity. In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid substrates with a binding affinity represented by an EC50 binding affinity. In some embodiments, the EC50 binding affinity is less than 500nM, 400nM, 300nM, 200nM, 100nM, 50nM, 40nM, 30nM, 20nM, 10nM, 5nM, 4nM, 3nM, 2nM, or 1.5nM. In some embodiments, the EC50 binding affinity is less than 10nM. In some embodiments, the EC50 binding affinity is less than 1.5nM. In some embodiments, the EC50 binding affinity is the same or different for different amyloid substrates. In some embodiments, the EC50 binding affinities are the same or different for human amyloid substrates.In some embodiments, the EC50 binding affinities are the same or different for the synthetic amyloid substrates.

[0235] 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 a target antigen. The components of the Fab region allow, for example, an antibody to bind to a specific ligand and further activate the immune system through that interaction. In the case of IgG, IgA, IgD, IgE, and IgM antibody isotypes, Ig is composed of two proteins, heavy and light chains, that interact in pairs to form an intact Ig that includes two heavy chains and two light chains. Both the heavy and light chains are further divided into variable and constant domains (the heavy chains forming the light and heavy variable domains that include the Fab functional region and the fragment crystallizable (Fc) domain that interacts with cell receptors and complement). The Fc region of Ig possesses highly conserved N-glycosylation sites.

[0236] In certain exemplary embodiments, one or more of the peptides shown in Table 1 below may be linked to an antibody or functional fragment thereof via the C- or N-terminus of the light chain protein or the C- or N-terminus of the heavy chain, thereby forming an antibody-peptide fusion protein. That is, any of the sequences identified below in Table 1 may be linked independently or simultaneously to the heavy or light chain of an antibody or functional fragment thereof to form a peptide-antibody conjugate. For example, two of the amyloid-reactive peptides may be linked to a single antibody, such as by connecting the amyloid-reactive peptide amino acid sequence to the N-terminus of an Ig light chain protein.

[0237] In certain exemplary embodiments, recombinant DNA technology may be used, where a nucleotide sequence encoding the peptide of the present invention is cloned and fused to an Ig light chain in an expression vector, transformed or transfected into a suitable host cell, and cultured under suitable conditions for expression. The peptide-Ig light chain fusion is then isolated. Advantageously, and as the skilled artisan will appreciate in view of the present disclosure, the methods described herein may be used to connect any peptide sequence to an antibody. That is, although an amyloid-reactive peptide is used as an example of a peptide linked to an antibody, the methods of linking a peptide to an antibody (e.g., the N- and / or C-terminus of a light chain protein and / or the N- and / or C-terminus of an Ig heavy chain protein) may be used to link a variety of different peptides to an antibody.

[0238] In certain exemplary embodiments, multiple identical or different peptides may be linked to a single antibody or functional fragment thereof. For example, a first expression vector may include a light chain nucleic acid sequence integrated with a nucleic acid sequence encoding peptide A, with the nucleic acid sequence for peptide A located within the vector, such that peptide A is expressed as linked to the N-terminus of the light chain protein. Additionally, a second expression vector may include a heavy chain nucleic acid sequence integrated with a nucleic acid sequence encoding peptide B, with the nucleic acid sequence for peptide B located within the vector, such that peptide B is expressed as linked to the N-terminus of the light chain protein.

[0239] In such an exemplary embodiment, when both expression vectors are expressed in the same cell, the resulting Ig protein may have one peptide A sequence on the N-terminus of each light chain (two peptide A in total) and peptide B on the N-terminus of the heavy chain. In certain exemplary embodiments, the vector may include peptide C on the C-terminus, resulting in an antibody with two peptide A sequences (one on each light chain), a peptide B sequence on the N-terminus of the heavy chain, and a peptide C sequence linked to the C-terminus of the heavy chain. Thus, and as one of skill in the art will understand based on this disclosure, expression vectors may be tailored to modify immunoglobulins to have the same or different combinations of proteins. As a specific example using an amyloid-reactive peptide, an antibody-peptide fusion protein may include two p5 protein sequences (SEQ ID NO:1) (i.e., one on each light chain N-terminus). In other exemplary embodiments, the peptide linked to the immunoglobulin may have affinity for a ligand and may be used to detect the ligand.

[0240] In some embodiments, the antibody-peptide fusion protein comprising the humanized antibody of the present disclosure comprises an Fc region. In some embodiments, the Fc is of IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the antibody-peptide fusion protein comprising the humanized antibody promotes Fc-mediated antibody effector function. In some embodiments, the antibody-peptide fusion protein comprising the humanized antibody promotes antibody-dependent cellular phagocytosis.

[0241] In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) that is less than about 100, 10, 1, 0.1, 0.01 μM. In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is less than about 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, or 100 μM (including any value or range between these values). In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is less than 500, 100, 10, or 1 nM. In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is less than about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 250, 500, 750, 1000, 2000, or 2200 nM. In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 250, 500, 750, 1000, 2000, or 2200 nM (including any value or range between these values). In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is about 40-50 nM. In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is 40-50 nM. In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is less than 50 nM. In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with a Kd that is less than the Kd of c11-1F4, which binds to human amyloid fibrils.

[0242] In some embodiments, the antibody-peptide fusion protein is administered at a concentration of antibody (EC 50In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with half-maximal binding at a concentration of antibody (EC) that is about 0.001, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 μM (including any value or range therebetween). 50 In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with half-maximal binding at a concentration of antibody (EC 50 In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with half-maximal binding at a concentration of antibody (EC) that is about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 100, 250, 500, 750, or 1000 nM (including any value or range therebetween). 50 In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with half-maximal binding at an antibody concentration (EC 50 In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with half-maximal binding at an antibody concentration (EC 50 In some embodiments, the antibody-peptide fusion protein binds to human amyloid fibrils with half-maximal binding at an EC of c11-1F4 that binds to human amyloid fibrils. 50 A concentration of antibody (EC 50 ) binds to human amyloid fibrils with half-maximal binding.

[0243] Dissociation constant and EC 50Methods for calculating the EC are known in the art and include, for example, surface plasmon resonance and europium-linked immunosorbent assay (EuLISA). In some embodiments, the dissociation constant is determined by measuring binding to Len(1-22) monomer peptide, for example, using surface plasmon resonance. In some embodiments, the EC 50 is determined using EuLISA. In some embodiments, EC 50 is determined using EuLISA to measure the level of binding to rVλ6Wil fibrils, Per125 wtATTR extracts, Ken ATTR extracts, SHI ALλ liver extracts, or TAL ALκ liver extracts.

[0244] In some embodiments, the antibody-peptide fusion protein is conjugated to a detectable label. In some embodiments, the detectable label is a radionuclide (e.g., I- 125 , I- 123 , I- 124 , I- 131 , Zr- 89 , Tc- 99m , Cu- 64 , Br- 76 , F- 18 ); enzymes (horseradish peroxidase); biotin; and fluorophores, etc. Any means known in the art for detectably labeling proteins may be used and / or adapted for use with the methods described herein. For example, the antibody-peptide fusion protein may be radiolabeled with a radioisotope, or labeled with a fluorescent or chemiluminescent tag. Exemplary radioisotopes include, for example, 18 F, 111 In, 99m Tc, and 123 I, and 125I. These and other radioisotopes can be attached to the antibody-peptide fusion protein using well-known chemistry that may or may not involve the use of chelators such as, for example, DTPA or DOTA covalently linked to the light chain protein of the antibody-peptide fusion protein. Exemplary fluorescent or chemiluminescent tags include fluorescein, Texas Red, rhodamine, Alexa dyes, and luciferase, which can be conjugated to the antibody-peptide fusion protein by reaction with lysine, cysteine, glutamic acid, and aspartic acid side chains. In one exemplary embodiment, the label is detected using a fluorescence microplate reader, or a fluorometer, using excitation and emission wavelengths appropriate for the tag used. Radioactive labels can be detected, for example, using a gamma or scintillation counter depending on the type of radioactive emission, and by using a suitable energy window for accurate detection of the particular radionuclide. However, any other suitable technique for detection of radioisotopes can also be used to detect the label. In some embodiments, the detectable label is 125 I.

[0245] In some embodiments, the antibody-peptide fusion protein binds to rVλ6Wil fibrils, Per125 wtATTR extracts, KEN hATTR extracts, SHI ALλ liver extracts, and / or TAL ALκ liver extracts. In some embodiments, the antibody-peptide fusion proteins described herein bind to amyloid deposits or fibrils. In some embodiments, the antibody-peptide fusion protein binds to one or more amyloidogenic peptides in amyloid. In some embodiments, the amyloid to which the antibody-peptide fusion protein binds comprises an amyloidogenic λ6 variable domain protein (Vλ6Wil) or an amyloidogenic immunoglobulin light chain (AL), Aβ(1-40) amyloid-like fibrils or an amyloidogenic Aβ precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid to which the antibody-peptide fusion protein binds is an amyloidogenic form of immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTR), apolipoprotein AI (AApoAI), apolipoprotein AII (AApoAII), gelsolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactadherin (AMed), islet amyloid polypeptide (AIAPP), Prolactin (APro), insulin (AIns), prion protein (APrP); alpha-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or IAAP, ALκ4, Alλ1, and other amyloidogenic peptides. The amyloidogenic peptide to which the antibody-peptide fusion protein binds can be a protein, protein fragment, or protein domain. In some embodiments, the amyloid deposits or amyloid fibrils comprise recombinant amyloidogenic proteins. In some embodiments, the amyloid is part of the pathology of a disease.

[0246] In some embodiments, binding of the antibody-peptide fusion protein to human amyloid promotes phagocytosis of human amyloid fibrils. In some embodiments, the antibody-peptide fusion protein opsonizes human amyloid fibrils. In some embodiments, the antibody-peptide fusion protein opsonizes rVλ6Wil fibrils. In some embodiments, contacting human amyloid fibrils with an antibody-peptide fusion protein of the present disclosure in the presence of macrophages promotes uptake of human amyloid fibrils by macrophages. In some embodiments, contacting human amyloid fibrils with an antibody-peptide fusion protein of the present disclosure in the presence of macrophages promotes opsonization of human amyloid fibrils. In some embodiments, binding of the antibody-peptide fusion protein to human amyloid fibrils promotes phagocytosis of human amyloid to an extent equal to or greater than control antibodies (e.g., hIgG and / or c11-1F4). In some embodiments, the antibody-peptide fusion protein promotes antibody-dependent cellular phagocytosis.

[0247] In some embodiments, the antibody-peptide fusion protein exhibits one or more in vivo characteristics selected from 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 a reference antibody, and the antibody-peptide fusion protein is detectable in organs throughout the body. In some embodiments, the antibody-peptide fusion protein exhibits improved biodistribution, and 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 a reference antibody, and the antibody-peptide fusion protein exhibits reactivity to one or more different amyloid substrates in vivo. In some embodiments, the antibody-peptide fusion protein is reactive to 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 improved phagocytosis compared to the reference antibody, and contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in an increased level of phagocytosis. In some embodiments, contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in clearance of the amyloid substrate. In some embodiments, contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in improved phagocytosis and clearance of the amyloid substrate. In some embodiments, contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in a therapeutic benefit for an individual having an amyloid-related disorder. In some embodiments, the reference antibody is not engineered to bind amyloid substrate. In some embodiments, the reference antibody does not comprise an 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 IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the reference antibody is an IgG1 isotype.

[0248] Also provided herein are compositions comprising an antibody-peptide fusion protein comprising an amyloid reactive peptide and an antibody that binds to human amyloid fibrils. 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 some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus of the heavy chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the heavy chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer. In some embodiments, the composition comprises an antibody-peptide fusion protein, and at least 80% of the antibody-peptide fusion protein is intact. In some embodiments, the composition comprises an antibody-peptide fusion protein, wherein at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% of the antibody-peptide fusion protein is intact. In some embodiments, an intact antibody-peptide fusion protein is one that has not been subjected to proteolytic degradation. In some embodiments, the intact antibody-peptide fusion protein consists of a full-length antibody-peptide fusion protein. In some embodiments, the intact antibody-peptide fusion protein comprises a full-length amino acid sequence selected from the group consisting of SEQ ID NOs: 87-92. In some embodiments, the antibody is a full-length antibody.

[0249] In some embodiments, the composition has a purity defined by the amount of intact antibody-peptide fusion protein present in the composition. In some embodiments, the intact antibody-peptide fusion protein has not been subjected to proteolytic degradation. In some embodiments, the intact antibody-peptide fusion protein consists of a full-length antibody-peptide fusion protein. In some embodiments, the intact antibody-peptide fusion protein comprises a full-length amino acid sequence selected from the group consisting of SEQ ID NOs: 87-92. In some embodiments, the composition has a purity of at least 80%. For example, a composition having a purity of 80% comprises 80% intact antibody-peptide fusion protein. In some embodiments, the composition has a purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. In some embodiments, the intact antibody-peptide fusion protein consists of a full-length antibody-peptide fusion protein. In some embodiments, the intact antibody-peptide fusion protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 87-92.

[0250] In some embodiments, the composition comprises an antibody-peptide fusion protein comprising an amyloid reactive peptide and an antibody that binds to human amyloid fibrils. 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 some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus of the heavy chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the heavy chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked to the antibody via a spacer or without a spacer.

[0251] In some embodiments, the compositions described herein comprise 20% or less cleavage products. In some embodiments, the cleavage products comprise a light chain lacking one or more amino acid residues from the N-terminus or C-terminus. In some embodiments, the cleavage products comprise a light chain comprising, in the N-terminus to C-terminus direction, a variable light chain region, a constant light chain region, a spacer, and an amyloid-reactive peptide. In some embodiments, the cleavage products comprise an amyloid-reactive peptide lacking one or more amino acid residues from the C-terminus. In some embodiments, the cleavage products comprise an antibody linked to an amyloid-reactive peptide, the antibody or the amyloid-reactive peptide lacking one or more residues at the N-terminus or C-terminus. In some embodiments, the cleavage products comprise a light chain comprising, in the N-terminus to C-terminus direction, an amyloid-reactive peptide, a spacer, a variable light chain region, and a constant light chain region. In some embodiments, the cleavage products comprise an amyloid-reactive peptide lacking one or more amino acid residues from the N-terminus. In some embodiments, the cleavage products comprise a heavy chain lacking one or more amino acid residues from the N-terminus or C-terminus. In some embodiments, the compositions described herein contain no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products.

[0252] In some embodiments, the compositions described herein comprise 20% or less cleavage products. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence shown by SEQ ID NO:87-90. In some embodiments, the cleavage products comprise light chains comprising an amyloid-reactive peptide lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence shown by SEQ ID NO:87. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence shown by SEQ ID NO:87-90. In some embodiments, the cleavage products comprise light chains comprising an amyloid-reactive peptide lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence shown by SEQ ID NO:89-90. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence shown by SEQ ID NO:91-92. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence shown by SEQ ID NO:91-92. In some embodiments, the cleavage products comprise heavy chains that comprise an amyloid-reactive peptide that is missing one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth by SEQ ID NO: 92. In some embodiments, the cleavage products comprise light or heavy chains that comprise an amyloid-reactive peptide that is missing one or more amino acid residues from the N-terminus or C-terminus. In some embodiments, the compositions described herein comprise no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products.

[0253] In some embodiments, the compositions described herein comprise 20% or less cleavage products. In some embodiments, the cleavage products comprise a light chain lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:89. In some embodiments, the cleavage products comprise a light chain lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:89. In some embodiments, the cleavage products comprise an amyloid-reactive peptide lacking one or more amino acid residues from the C-terminus. In some embodiments, the cleavage products comprise an antibody fused to an amyloid-reactive peptide, the amyloid-reactive peptide being truncated by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 amino acids. In some embodiments, the cleavage products comprise a heavy chain lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the cleavage products lack at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 15 amino acids at the N-terminus compared to the intact fusion protein. In some embodiments, the cleavage products comprise a heavy chain lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO: 91. In some embodiments, the compositions described herein comprise no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products.

[0254] In some embodiments, the compositions described herein comprise no more than 20% cleavage products. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:87. In some embodiments, the cleavage products comprise amyloid-reactive peptides lacking one or more amino acid residues from the N-terminus. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:87. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the compositions described herein comprise no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products.

[0255] In some embodiments, the compositions described herein comprise no more than 20% cleavage products. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:88. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:88. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:92. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:92. In some embodiments, the cleavage products comprise amyloid-reactive peptides lacking one or more amino acid residues from the C-terminus. In some embodiments, the compositions described herein comprise no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products.

[0256] In some embodiments, the compositions described herein comprise no more than 20% cleavage products. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:90. In some embodiments, the cleavage products comprise light chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:90. In some embodiments, the cleavage products comprise amyloid-reactive peptides lacking one or more amino acid residues from the C-terminus. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the N-terminus compared to the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the cleavage products comprise heavy chains lacking one or more amino acid residues from the C-terminus compared to the amino acid sequence set forth in SEQ ID NO:91. In some embodiments, the compositions described herein comprise no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products.

[0257] In some embodiments, the composition comprises an antibody-peptide fusion protein, and the antibody-peptide fusion protein exhibits EC50 binding affinity to one or more amyloid substrates. In some embodiments, the EC50 binding affinity is less than 500nM, 400nM, 300nM, 200nM, 100nM, 50nM, 40nM, 30nM, 20nM, 10nM, 5nM, 4nM, 3nM, 2nM, or 1.5nM. In some embodiments, the EC50 binding affinity is less than about 100nM or 100nM. In some embodiments, the EC50 binding affinity is less than about 10nM or 10nM. In some embodiments, the EC50 binding affinity is less than about 1.5nM or 1.5nM. 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 affinities are the same or different for the synthetic amyloid substrates.

[0258] In some embodiments, the composition comprises an antibody-peptide fusion protein, the antibody-peptide fusion protein exhibits one or more in vivo characteristics selected from 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 a reference antibody, the antibody-peptide fusion protein is detectable in organs throughout the body. In some embodiments, the antibody-peptide fusion protein exhibits improved biodistribution, 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 a reference antibody, the antibody-peptide fusion protein exhibits reactivity to one or more different amyloid substrates in vivo. In some embodiments, the antibody-peptide fusion protein is reactive to 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 improved phagocytosis compared to the reference antibody, and contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in an increased level of phagocytosis. In some embodiments, contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in clearance of the amyloid substrate. In some embodiments, contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in improved phagocytosis and clearance of the amyloid substrate. In some embodiments, contacting an amyloid substrate with the antibody-peptide fusion protein in vivo results in a therapeutic benefit for an individual having an amyloid-related disorder. In some embodiments, the reference antibody does not bind the amyloid substrate. In some embodiments, the reference antibody does not comprise an amyloid-reactive peptide. In some embodiments, the reference antibody is an IgG antibody. In some embodiments, the reference antibody is an IgG1, IgG2, IgG3, or IgG4 isotype.In some embodiments, the reference antibody is of the IgG1 isotype.

[0259] In some embodiments, the composition further comprises a pharma- ceutically acceptable carrier, which in some embodiments may be a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, or a combination thereof.

[0260] In various embodiments, compositions according to the present disclosure may be formulated for delivery via any route of administration, including, for example, aerosol, nasal, oral, transmucosal, transdermal, parenteral, or enteral. In some embodiments, administration is intravenous or subcutaneous.

[0261] Pharmaceutical formulations are provided that include the antibody-peptide fusion protein antibodies. Pharmaceutical compositions and formulations typically include one or more optional pharma- ceutically acceptable carriers or excipients. In some embodiments, the compositions include at least one additional therapeutic agent.

[0262] The term "pharmaceutical formulation" refers to a preparation which is in a form such that the biological activity of the active ingredients contained therein is effective, and which does not contain additional ingredients that are unacceptably toxic to a subject to which the formulation will be administered.

[0263] "Pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0264] A buffering agent is included in the composition in some embodiments. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some embodiments, 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, for example, in Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0265] Also provided herein is a pharmaceutical composition comprising any of the antibody-peptide fusion proteins described herein. In some embodiments, the pharmaceutical composition further comprises a pharma- ceutically acceptable carrier.

[0266] In certain exemplary embodiments, the antibody-peptide fusion protein may be obtained by isolation or purification. Protein purification techniques involve, at one level, homogenization and crude fractionation of cells, tissues, or organs into peptide and non-peptide fractions. Other protein purification techniques include precipitation, for example, with ammonium sulfate, polyethylene glycol (PEG), antibodies, etc., or by heat denaturation, followed by: centrifugation; chromatographic steps, such as ion exchange, gel filtration, reverse phase, hydroxylapatite, and affinity chromatography; isoelectric focusing; gel electrophoresis, such as polyacrylamide gel electrophoresis; and combinations of these and other techniques.

[0267] Various chromatographic techniques include, but are not limited to, ion exchange chromatography, gel exclusion chromatography, affinity chromatography, immunoaffinity chromatography, and reverse phase chromatography.A particularly efficient method for purifying peptides is fast performance liquid chromatography (FPLC) or even high performance liquid chromatography (HPLC).In certain exemplary embodiments, the Fc domain can be linked to the amyloid-reactive peptide via a GGGYS linker sequence (SEQ ID NO:27).

[0268] III. Diagnostic and Detection Methods Also provided herein are methods of identifying amyloid deposits in a subject.

[0269] In some embodiments, provided herein is a method of identifying amyloid deposits in a subject, comprising administering to the subject any one of the antibody-peptide fusion proteins described herein, where the antibody-peptide fusion protein comprises a detectable label, and detecting a signal from the antibody-peptide fusion protein. Any one of the detectably labeled antibody-peptide fusion proteins described herein may be used. In some embodiments, the subject is determined to be amyloid-free or to be afflicted with monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), or one or more related plasma cell dyscrasias. In some embodiments, the antibody-peptide fusion protein comprises an amyloid-reactive peptide comprising the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:2. In some embodiments, the antibody-peptide fusion protein comprises an antibody that binds to human amyloid fibrils. In some embodiments, the antibody comprises a variable heavy chain (VH) and a variable light chain (VL), 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, and 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. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus or C-terminus of the light chain. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus or C-terminus of the heavy chain. In some embodiments, the amyloid reactive peptide is linked to the antibody via a spacer. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 23-24, 27, and 83-86. In some embodiments, the spacer comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-86. In some embodiments, the spacer comprises the amino acid sequence set forth in SEQ ID NO: 83. In some embodiments, the antibody is a full-length antibody.

[0270] In some embodiments, provided herein is a method of detecting a ligand comprising contacting the ligand with any one of the antibody-peptide fusion proteins described herein, where the antibody-peptide fusion protein comprises a detectable label, and where a peptide of the antibody-peptide fusion protein has binding affinity for the ligand, and determining a signal from the detectable label, thereby detecting the ligand. Any one of the detectably labeled antibody-peptide fusion proteins described herein may be used.

[0271] In some embodiments, the antibody-peptide fusion proteins can be labeled with various agents to allow for their detection in in vivo and in vitro assays, for example after the fusion peptide has been purified. This includes, but is not limited to, radionuclides (e.g., I- 125 , I- 123 , I- 131 , Zr- 89 , Tc- 99m , Cu- 64 , Br- 76 , F- 18 ); enzymes (horseradish peroxidase); biotin; fluorophores, and the like. Any means known in the art for detectably labeling proteins may be used and / or adapted for use in the methods described herein. For example, antibodies or fragments thereof, and / or amyloid-reactive peptides may be radiolabeled with a radioisotope, or labeled with a fluorescent or chemiluminescent tag. Exemplary radioisotopes include, for example, 18 F, 111 In, 99m Tc, and 123 I, and 125I. These and other radioisotopes can be attached to isolated immunoglobulin light chains using well-known chemistry that may or may not involve the use of chelators such as DTPA or DOTA covalently linked to the light chain protein of the antibody. Exemplary fluorescent or chemiluminescent tags include fluorescein, Texas Red, rhodamine, Alexa dyes, and luciferase, which can be conjugated to proteins by reaction with lysine, cysteine, glutamic acid, and aspartic acid side chains. In one exemplary embodiment, the label is detected using a fluorescence microplate reader, or a fluorometer, using excitation and emission wavelengths appropriate for the tag used. Radioactive labels can be detected using, for example, a gamma or scintillation counter by using an energy window suitable for the type of radioactive emission and for accurate detection of the particular radionuclide. However, any other suitable technique for detection of radioisotopes can also be used to detect the label.

[0272] With respect to amyloidosis, such labels may be used, for example, to diagnose the presence of amyloid, to determine amyloid protein load, to monitor the ability of the antibody-peptide fusion protein to bind amyloid in a particular subject, to monitor the progression of amyloidosis, and / or to monitor the subject's response to an amyloid treatment (including treatment involving administration of the antibody-peptide fusion protein to the subject). For example, an antibody-peptide fusion protein may be labeled with a detectable label as described herein and then administered to a subject suffering from or suspected of suffering from an amyloid-based disease (e.g., amyloidosis, monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), or related plasma cell dyscrasias). The subject may then be imaged, for example, to detect the presence of the antibody-peptide fusion protein.

[0273] In certain exemplary embodiments, the signal from the detectably labeled antibody-peptide fusion protein can be quantified, thereby providing an indication of the level of amyloid deposits in the subject. For example, the signal intensity can be compared to a standard signal threshold, and above the threshold, amyloidosis is present, while below the threshold, amyloidosis is absent or at a low level. The subject can be diagnosed as having amyloid, in which case treatments such as chemotherapy, corticosteroid drugs (lenalidomide or thalidomide) and / or bortezomib (Velcade) can be administered. Additionally or alternatively, the antibody-peptide fusion proteins described herein can be administered to the subject in an effort to treat the subject as described herein. In certain exemplary embodiments, the subject can be stratified into one or more groups, e.g., low amyloid burden, medium amyloid burden, or high amyloid burden, and then treated accordingly. To monitor treatment progress, the subject can be re-administered the antibody-peptide fusion protein, thereby re-evaluating their amyloid burden.

[0274] IV. Treatment Method Also provided herein is a method 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.

[0275] In some embodiments, a method of treating an amyloid disease (e.g., amyloidosis) is provided, comprising administering to a subject in need thereof a therapeutically effective amount of any one of the antibody-peptide fusion proteins described herein.

[0276] In other embodiments, the amyloidosis is systemic amyloidosis. In some embodiments, the amyloidosis is familial amyloidosis. In other embodiments, the amyloidosis is sporadic amyloidosis. In some embodiments, the amyloidosis or amyloid-related disease is AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, hATTR amyloidosis, ALect2 amyloidosis, and IAPP amyloidosis of type II diabetes, Alzheimer's disease, Down's syndrome, Hereditary cerebral hemorrhage with amyloidosis of the Dutch type, cerebral beta-amyloid angiopathy, spongiform encephalopathy, thyroid tumors, Parkinson's disease, dementia with Lewy bodies, tauopathy, Huntington's disease, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, age-related pituitary disorders, iatrogenic syndromes, spongiform encephalopathies, reactive chronic inflammation, thyroid tumors, myeloma or other forms of cancer. In some embodiments, the amyloid-related disease is selected from the group consisting of AL, AH, Aβ2M, ATTR, transthyretin, AA, AApoAI, AApoAII, AApoAIV, AApoCII, AApoCII, AGel, ALys, ALEct2, AFib, ACys, ACal, AMed, AIAPP, APro, AIns, APrP, ASPC, AGal7, ACor, Aker, ALac, AOAPP, ASem1, AEnf, or Aβ amyloidosis. In some embodiments, treatment with the antibody-peptide fusion protein results in clearance of amyloid. In some embodiments, the antibody-peptide fusion protein binds to amyloid associated with normal aging. In other embodiments, the antibody-peptide fusion protein is used in the diagnosis, treatment, or prognosis of amyloidosis or amyloid-related disease in a subject.

[0277] In some embodiments, the amyloid-related disease is localized amyloidosis.

[0278] In some embodiments, the antibody-peptide fusion protein is administered via intradermal, subcutaneous, intramuscular, intracardiac, intravascular, intravenous, intraocular, intraarterial, epidural, intrathecal, extracorporeal, intrathecal, intraperitoneal, intrapleural, intraluminal, intravitreal, intracavernous, intraventricular, intraosseous, intraarticular, intracellular, or pulmonary routes.

[0279] In some embodiments, the antibody-peptide fusion protein is administered in an amount sufficient to induce phagocytosis of amyloid by cells of the immune system (eg, macrophages).

[0280] In some embodiments, the subject is a mammal, such as a primate, cow, rodent, or pig, hi some embodiments, the subject is a human.

[0281] Also provided herein are methods of targeting amyloid deposits for clearance. In some embodiments, the methods comprise contacting the amyloid deposits with an antibody-peptide fusion protein of the present disclosure.

[0282] In some embodiments, amyloid deposits may contribute to disease pathology. In other embodiments, amyloid deposits may indicate amyloidosis or amyloid-related disease in a subject. In some embodiments, the antibody-peptide fusion protein binds to amyloid in a subject with amyloidosis. In some embodiments, amyloidosis is localized to a specific tissue or organ system, such as the liver, heart, or central nervous system.

[0283] In some embodiments, amyloid deposits are cleared. In some embodiments, amyloid deposits are cleared. In some embodiments, amyloid deposits are opsonized by the antibody-peptide fusion protein. In some embodiments, binding of the antibody-peptide fusion protein to human amyloid fibrils promotes phagocytosis of human amyloid fibrils and removal of amyloid deposits. In some embodiments, the antibody-peptide fusion protein opsonizes human amyloid fibrils, thereby removing amyloid deposits. In some embodiments, the antibody-peptide fusion protein opsonizes rVλ6Wil fibrils. In some embodiments, binding of the antibody-peptide fusion protein to human amyloid fibrils promotes phagocytosis and / or opsonization of human amyloid fibrils to an extent equal to or greater than control antibodies (e.g., mIgp5 and / or c11-1F4).

[0284] In some embodiments, provided herein are methods of treating an amyloid-related disorder, comprising administering an antibody-peptide fusion protein conjugated to a detectable label, detecting the label, and administering an amyloidosis treatment to a subject if a signal is detected. In some embodiments, the detectable label is a radioactive label. In some embodiments, the detectable label is an I 125 , Tc 99 In some embodiments, the detectable label is a label. In some embodiments, the detectable label is a fluorescent label. In some embodiments, the detectable label is an enzyme label. In some embodiments, the label is horseradish peroxidase or alkaline phosphatase. The label further comprises a chemical moiety, such as a specific associated detectable moiety, for example, biotin, that can be detected via binding to labeled avidin. In some embodiments, amyloid deposits are identified in the liver, spleen, or blood of the subject. In some embodiments, the amyloidosis treatment comprises an antibody-peptide fusion protein provided herein.

[0285] Also provided herein are methods of identifying amyloid deposits in a subject, comprising administering an antibody-peptide fusion protein, wherein the antibody-peptide fusion protein is conjugated to a detectable label. In some embodiments, the method comprises detecting a signal from the antibody-peptide fusion protein. In some embodiments, the detectable label is a radioactive label. In some embodiments, the detectable label is an I 125 , Tc 99 In some embodiments, the detectable label is a fluorescent label. In some embodiments, the detectable label is an enzyme label. In some embodiments, the label is horseradish peroxidase or alkaline phosphatase. The label further comprises a specific associated detectable moiety, such as biotin, that can be detected via binding to labeled avidin. In some embodiments, amyloid deposits are identified in the liver, spleen, or blood of the subject.

[0286] In some embodiments, provided herein are methods of detecting a ligand, comprising contacting the ligand with an antibody-peptide fusion conjugated to a detectable label and determining a signal from the detectable label. In some embodiments, the detectable label is a radioactive label. In some embodiments, the detectable label is an I 125 , Tc 99 In some embodiments, the detectable label is a label. In some embodiments, the detectable label is a fluorescent label. In some embodiments, the detectable label is an enzyme label. In some embodiments, the label is horseradish peroxidase or alkaline phosphatase. The label further comprises a specific associated detectable moiety, such as biotin, that can be detected via binding to labeled avidin. In some embodiments, the detection is in vitro. In some embodiments, the detection is in vivo.

[0287] V. Nucleic Acids, Vectors, Host Cells, and Methods for Producing Peptide-Antibody Fusion Proteins Also provided herein is a nucleic acid(s) encoding the antibody-peptide fusion protein of the disclosure. The antibody-peptide fusion protein can be any of the antibody-peptide fusion proteins described herein.

[0288] In some embodiments, the nucleic acids provided herein are in one or more vectors. For example, in some embodiments, vectors are provided herein that include an antibody heavy chain and a light chain, where the light chain is linked to a peptide. In some embodiments, the heavy chain and the light chain linked to the peptide are in different vectors.

[0289] In some embodiments, the vector comprises a nucleic acid(s) encoding an antibody-peptide fusion protein of the disclosure.

[0290] For antibody production, the heavy and light chains linked to the peptide expression vectors can be introduced into a suitable production cell line known in the art. Introduction of the expression vectors can be accomplished by co-transfection via electroporation or any other suitable transformation technique available in the art. The antibody-producing cell line is then selected and grown, and the antibody is purified. The purified antibody can then be analyzed by standard techniques, such as SDS-PAGE.

[0291] Also provided are host cells comprising nucleic acids encoding any of the antibody-peptide fusion proteins described herein. Suitable host cells for cloning or expressing vectors encoding antibodies include prokaryotic or eukaryotic cells described herein. For example, antibody-peptide fusion proteins can be produced in bacteria, particularly when glycosylation and Fc effector functions are not required. 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 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, which describes expression of antibody fragments in E. coli.) After expression, the peptide-linked antibody can be isolated from the bacterial cell paste in a soluble fraction and further purified.

[0292] In some embodiments, a host cell comprising a vector comprising a nucleic acid(s) encoding an antibody-peptide fusion protein of the disclosure.

[0293] Suitable host cells for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plants and insect cells. In particular, a number of baculovirus strains have been identified that can be used in conjunction with insect cells for transfection of Spodoptera frugiperda cells.

[0294] Plant cell cultures may also be utilized as hosts, see, e.g., U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing the PLANTIBODIES™ technology for producing antibodies in transgenic plants).

[0295] Vertebrate cells may also be used as hosts. For example, mammalian cell lines adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines include monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney lines (293 cells or 293 cells described, for example, in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse Sertoli cells (for example, TM4 cells described, for example, 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 hepatocytes (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); for example, Mather et al., Annals N Y. Acad. Sci. 383:44-68 (1982); MRC5 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, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

[0296] VI. Purification method Also provided herein are methods of producing an antibody-peptide fusion protein of the disclosure, in some embodiments, the method comprises culturing a host cell of the disclosure under conditions suitable for expression of a vector encoding the antibody-peptide fusion protein and recovering the antibody-peptide fusion protein.

[0297] In some embodiments, a method for producing an antibody-peptide fusion protein comprises: i) culturing a host cell comprising a vector encoding the antibody-peptide fusion protein under perfusion cell culture conditions suitable for expression of the antibody-peptide fusion protein; and ii) harvesting 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 that binds to human amyloid fibrils, the antibody comprising a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL). In some embodiments, the amyloid-reactive peptide and the antibody are linked at the N-terminus or C-terminus of the light chain, and the amyloid-reactive peptide is linked to the antibody with or without a spacer. In some embodiments, the amyloid-reactive peptide and the antibody are linked at the N-terminus or C-terminus of the heavy chain, and the amyloid-reactive peptide is linked to the antibody with or without a spacer. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer, hi some embodiments, the antibody is a full-length antibody.

[0298] In some embodiments, the method includes culturing the host cells using a fed-batch method. Fed-batch culture refers to a method of culturing cells in which the cell culture is replenished with fresh medium, i.e., the cells are "fed" with new medium, but the spent medium is not removed. Typically, a "fed-batch" culture process is carried out in a bioreactor, and additional components (e.g., nutrient supplements) are added to the culture at some time after the start of the culture process. The controlled addition of nutrients directly affects the growth rate of the culture and allows the avoidance of the build-up of overflow metabolites (see, e.g., Wlaschin, KF 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 stopped at some point and the cells and / or components in the medium are harvested and optionally purified.

[0299] In some embodiments, the method includes culturing the host cells using a perfusion culture method. Perfusion refers to a method of culturing cells in which 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 be performed continuously or intermittently, as desired, for a period of time. The fresh medium added during perfusion typically provides nutritional supplements for cells that are depleted during the culturing process. Perfusion also allows for the removal of cell consumption products and toxic by-products from the cell culture. Perfusion is performed during the cell growth phase and can be continued after the cells are transferred to fed-batch cell culture.

[0300] In some embodiments, the method of producing an antibody-peptide fusion protein comprises culturing a host cell comprising a vector encoding the 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 amyloid reactive peptide and an antibody that binds to human amyloid fibrils, the antibody comprising a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL). In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus or C-terminus of the light chain, and the amyloid reactive peptide is linked to the antibody through or without a spacer. In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus or C-terminus of the heavy chain, and the amyloid reactive peptide is linked to the antibody through or without a spacer. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer, hi some embodiments, the antibody is a full-length antibody.

[0301] In some embodiments, the method of producing the antibody-peptide fusion protein further comprises a continuous cell culture method. In some embodiments, the method comprises culturing the cells under perfusion conditions. In some embodiments, the method comprises harvesting the antibody-peptide fusion protein about every 12-36 hours. In some embodiments, the antibody-peptide fusion protein is harvested about every 12-24 hours, and the antibody-peptide fusion protein is harvested about every 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours. In some embodiments, the antibody-peptide fusion protein is harvested about every 24-36 hours, and the antibody-peptide fusion protein is harvested about every 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 hours. In some embodiments, the antibody-peptide fusion protein is harvested about every 12-16 hours, 14-18 hours, 16-20 hours, 16-24 hours, 20-32 hours, 18-36 hours, 24-32 hours, or 16-28 hours. In some embodiments, the antibody-peptide fusion protein is harvested after 12, 16, 20, 24, 28, 32, or 36 hours or less. In some embodiments, the antibody-peptide fusion protein is harvested about every day. In some embodiments, the antibody-peptide fusion protein is harvested about every 1, 2, 3, 4, 5, 6, or 7 days. In some embodiments, the antibody-peptide fusion protein is harvested about every 1-4 days, 2-5 days, 3-6 days, 1-5 days, 3-7 days, or 1-7 days. In some embodiments, the antibody-peptide fusion protein is harvested after 1, 2, 3, 4, 5, 6, or 7 days or less. In some embodiments, the antibody-peptide fusion protein comprises an amyloid reactive peptide and an antibody that binds to human amyloid fibrils, the antibody comprising a heavy chain comprising a heavy chain variable region (VH) and a light chain comprising a light chain variable region (VL). In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus or C-terminus of the light chain, and the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer.In some embodiments, the amyloid reactive peptide and the antibody are linked at the N-terminus or C-terminus of the heavy chain, and the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer. In some embodiments, the amyloid reactive peptide and the antibody are linked at the C-terminus of the light chain, and the amyloid reactive peptide is linked to the antibody via a spacer or without a spacer. In some embodiments, the antibody is a full-length antibody.

[0302] In some embodiments, the method of producing an antibody-peptide fusion protein further comprises applying the antibody-peptide fusion recovered in the recovering step to a cation exchange chromatography column. In some embodiments, the cation exchange chromatography column is used to isolate the intact antibody-peptide fusion protein. In some embodiments, the cation exchange chromatography column is used to separate the truncated antibody-peptide fusion protein from the intact antibody-peptide fusion protein. In some embodiments, the recovered antibody-peptide fusion is applied to the cation exchange chromatography column. In some embodiments, the cation exchange chromatography column is washed with one or more buffers comprising dissolved salts. In some embodiments, the cation exchange chromatography column is washed with a low salt buffer. In some embodiments, the cation exchange chromatography column is washed with a high salt buffer. In some embodiments, the cation exchange chromatography column is first washed with a low salt buffer, followed by application of a high salt buffer elution buffer. In some embodiments, the low salt buffer elutes the truncated antibody-peptide fusion protein from the cation exchange chromatography column. In some embodiments, the high salt buffer elutes the intact antibody-peptide fusion protein from the cation exchange chromatography column. In some embodiments, the method comprises applying the antibody-peptide fusion protein to a cation exchange chromatography column, then washing the column with a first buffer containing a low salt concentration, and then eluting the intact antibody-peptide fusion protein with a second buffer containing a high salt concentration. In some embodiments, the intact antibody-peptide fusion protein is eluted separately from the truncated antibody-peptide fusion protein. In some embodiments, the method results in enrichment of the intact antibody-peptide fusion protein. In some embodiments, the method results in removal of the truncated antibody-peptide fusion protein from the isolated intact antibody-peptide fusion protein.In some embodiments, the method comprises applying the antibody-peptide fusion protein to a cation exchange chromatography column, then washing the column with a first buffer containing a low salt concentration, then applying a second buffer containing a high salt concentration, resulting in the isolation of the intact antibody-peptide fusion protein.

[0303] In some embodiments, the method comprises applying the antibody-peptide fusion protein to a cation exchange chromatography column, then washing the column with a first buffer containing a low salt concentration, followed by washing with a second buffer containing a high salt concentration, resulting in the isolation of the intact antibody-peptide fusion protein. In some embodiments, washing the column with the first buffer containing a low salt concentration elutes the truncated antibody-peptide fusion protein from the cation exchange chromatography column. In some embodiments, the first buffer containing a low salt concentration comprises a salt concentration of 200 mM, 210 mM, 220 mM, 230 mM, 240 mM, 250 mM, 260 mM, 270 mM, 280 mM, 290 mM, or 300 mM or less. In some embodiments, the first buffer comprising a low salt concentration comprises a salt concentration of 200 mM to 300 mM, 210 mM to 280 mM, 220 mM to 260 mM, 210 mM to 290 mM, 240 mM to 270 mM, or 230 mM to 260 mM. In some embodiments, the first buffer comprising a low salt concentration comprises a salt concentration that is about 220 mM, 240 mM, 260 mM, or 280 mM, or any range between these values. In some embodiments, the first buffer comprises a salt concentration of about 200 mM to about 300 mM, or about 220 mM to about 280 mM. In some embodiments, the first buffer comprising a low salt concentration comprises a salt concentration that is about 260 mM. In some embodiments, washing the column with a first buffer containing a low salt concentration followed by washing the column with a second buffer containing a high salt concentration elutes the intact antibody-peptide fusion protein from the cation exchange chromatography column. In some embodiments, the second buffer containing a high salt concentration comprises a concentration of salt of 350 mM, 360 mM, 370 mM, 380 mM, 390 mM, 400 mM, 410 mM, 420 mM, 430 mM, 440 mM, or 450 mM or greater. In some embodiments, the second buffer containing a high salt concentration comprises a concentration of salt of 350 mM to 450 mM, 360 mM to 430 mM, 370 mM to 410 mM, 360 mM to 440 mM, 390 mM to 420 mM, or 380 mM to 410 mM, or any range between these points.In some embodiments, the second buffer comprises a salt concentration of 350 mM to 450 mM or 380 mM to 420 mM. In some embodiments, the first buffer comprising a low salt concentration comprises a salt concentration that is about 380 mM, 400 mM, 420 mM, or 440 mM. In some embodiments, the first buffer comprising a low salt concentration comprises a salt concentration that is about 400 mM. In some embodiments, the pH of the first and second buffers is 4.5 to 6.5, 4.7 to 6.3, 5.1 to 5.7, 4.7 to 6.1, 5.5 to 5.9, or 5.3 to 5.7. In some embodiments, the pH of the first and second buffers is about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0. In some embodiments, the pH of the first and second buffer solutions is about 5.5. In some embodiments, the pH of the first and second buffer solutions is the same. In some embodiments, the pH of the first and second buffer solutions is different.

[0304] In some embodiments, the method of producing comprises applying the antibody-peptide fusion protein to a cation exchange chromatography column, then washing the column with a first buffer containing a low salt concentration, then eluting with a second buffer containing a high salt concentration, resulting in the isolation of the intact antibody-peptide fusion protein. In some embodiments, the method comprises applying the antibody-peptide fusion protein to a cation exchange chromatography column at a loading density, where loading density is defined as grams of antibody-peptide fusion protein per liter of resin (g / L). In some embodiments, the loading density is equal to or less than 20, 25, 30, 35, 40, 45, 50, or 55 g / L, or any range between these points. In some embodiments, the loading density is at least 15, 20, 25, 30, 35, 40, 45, or 50 g / L. In some embodiments, the loading density is 15-55, 20-45, 25-50, 20-35, 30-40, 35-50, or 20-50 g / L. In some embodiments, the loading density is 50 g / L.

[0305] In some embodiments, the method of producing 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.

[0306] In some embodiments, the method of generating 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 protein purity, including quantitative analysis of monoclonal antibodies. An antibody sample is mixed with an exchangeable SDS-gel buffer and then electrophoresed through an SDS-gel-filled capillary. The sample is injected into the capillary port using high voltage. Protein translocation through the separation matrix occurs in the 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 including 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).

[0307] In some embodiments, the method further comprises determining the purity, and 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 defined by the amount of intact antibody-peptide fusion protein present. In some embodiments, the intact antibody-peptide fusion protein consists of a full-length antibody-peptide fusion. In some embodiments, the antibody-peptide fusion protein has a purity of at least 80%. In some embodiments, the antibody-peptide fusion protein has a purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. In some embodiments, the antibody-peptide fusion protein has a purity of 80%-100%, 82%-96%, 85%-90%, 84%-92%, 80%-88%, 90%-98%, or 95%-100%. In some embodiments, antibody-peptide fusion protein purity positively correlates with amyloid substrate binding.

[0308] In some embodiments, the method further comprises determining the purity, and 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 defined by the amount of intact antibody-peptide fusion protein present. In some embodiments, the intact antibody-peptide fusion protein consists of a full-length antibody-peptide fusion. In some embodiments, the antibody-peptide fusion protein comprises no more than 20% cleavage products. In some embodiments, the cleavage products comprise a light chain comprising a light chain lacking one or more amino acid residues from the N-terminus or C-terminus. In some embodiments, the cleavage products comprise a heavy chain comprising a heavy chain lacking one or more amino acid residues from the N-terminus or C-terminus. In some embodiments, the antibody-peptide fusion protein comprises no more than 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% cleavage products. In some embodiments, the antibody-peptide fusion protein contains between 20% and 0%, between 15% and 2%, between 12% and 4%, between 15% and 8%, between 10% and 1%, between 5% and 2%, or between 2% and 0% cleavage products.

[0309] VII. Kit The present disclosure also provides a kit that contains the antibody-peptide fusion protein of the present disclosure.The kit of the present disclosure may include one or more containers that contain purified antibody-peptide fusion protein.In some embodiments, the kit further includes instructions for use according to the method of the present disclosure.In some embodiments, these instructions include the description of administration of the antibody-peptide fusion protein described herein to treat amyloid disease according to any method of the present disclosure.

[0310] In some embodiments, the instructions include instructions on how to detect amyloid deposits, e.g., in a subject, in a tissue sample, or in a cell. The kit may further include instructions for selecting an individual suitable for treatment based on identifying whether the individual has an amyloid disease described herein.

[0311] The label or package insert indicates that the composition is used for treating, for example, a disease of the disclosure. Instructions can be provided for practicing any of the methods described herein. VIII. EXAMPLES

[0312] The following examples further illustrate the present invention but should not be construed as limiting its scope in any way. In view of the present disclosure and the ordinary level of skill in the art, those skilled in the art will understand that the following examples are intended to be illustrative only and that numerous changes, modifications, and variations can be used without departing from the scope of the subject matter of the present disclosure. The accompanying figures are meant to be considered as an integral part of the specification and description of the present disclosure.

[0313] Example 1. Design of antibody-peptide fusion proteins The following examples describe the design of exemplary amyloid reactive peptide-antibody fusion protein constructs. Exemplary construct structures are provided in Figures 1-4. The amino acid sequences of the constructs are provided below in Tables E1 and E2. In Tables E1-E2, the amino acid sequence of the amyloid reactive peptide p5R is shown in bold, and the spacer sequence is underlined and italicized. [Table 9] [Table 10-1] [Table 10-2]

[0314] Example 2. Binding affinity of humanized anti-amyloid antibodies to Len(1-22) monomer peptide The binding affinity of the humanized anti-amyloid antibodies was determined, for example, as described in International Application No. PCT / US2020 / 060596. Table E3 below provides the results of a surface plasmon resonance (SPR) assay measuring the binding of the humanized anti-amyloid antibodies to the Len(1-22) monomer peptide. Specifically, humanized VH9 and VL4 sequences were tested with and without additional amino acid substitutions, as indicated in the "Ligand" column of Table E3. [Table 11]

[0315] Example 3. Binding affinity of humanized anti-amyloid antibodies to rVλ6Wil fibrils The ability of the humanized anti-amyloid antibodies to bind rVλ6Wil fibrils was tested in comparison with the chimeric antibody c11-1F4, e.g., by europium-linked immunosorbent assay (EuLISA) (Figure 5A). Based on the data shown in Figure 5A, c11-1F4 had an EC of approximately 72 nM. 50 VH10 / VL4 bound at an EC value of 17 nM. 50 VH9 / VL4 binds with an EC value of 7 nM 50 VH8 / VL4 bound at an EC value of 16 nM. 50 VH7 / VL4 bound at an EC value of 75 nM. 50 VH6 / VL3 binds with an EC value of 95 nM 50 VH9 / VL4 showed increased binding compared to c11-1F4 (FIG. 5A). The VH9 / VL4 antibody bound amyloid fibrils to a greater extent than VH6 / VL3 (FIG. 5A).

[0316] As further described in PCT / US2020 / 060596 (incorporated herein by reference in its entirety), peptides p5 and p5R were added to the N-terminus of the light chain of a humanized antibody having VH9 / VL4 and VH6 / VL3. As shown in Figure 5B, addition of peptides p5 and p5R to the N-terminus of the light chain of VH6 / VL3 improved binding to rVλ6Wil fibrils by approximately 30-fold (EC50 Based on the data shown in Figure 5B, VH6 / VL3-p5 has an EC 50 VH6 / VL3-p5R bound with an EC value of 3 nM. 50 value, and c11-1F4 has an EC 50 VH6 / VL3 binds with an EC value of 95 nM 50 Combined by value.

[0317] Overall, the variant with the arginine variant of p5 (p5R) was superior to the p5 variant (FIGS. 5C and 5D).

[0318] As shown in FIG. 5E, VH9 / VL4 had the same reactivity to rVλ6Wil fibrils as the murine parent.

[0319] As shown in Figures 5E and 5F, both VH6 / VL3-p5 and VH6 / VL3-p5R show binding to hATTR amyloid extracts. Based on the data in Figures 5E and 5F, VH6 / VL3-p5 has an EC 50 Sno with hATTR extracts and an EC value of 90 nM 50 VH6 / VL3-p5R bound to Ken ATTR extract with an EC value of 47 nM. 50 EC value of 70 nM for Sno ATTR extracts 50 EC values ​​of 85 nM and 85 nM for Ken ATTR extracts. 50 Per125 bound to wtATTR at 100 ng / mL.

[0320] Table E4 below provides the results of EuLISA measuring the ability of mIgp5, hIgG1, c11-1F4, m11-1F4, VH6 / VL3-p5, VH9 / VL4-p5, and VH9 / VL4-p5R to bind rVλ6Wil fibrils, Per125 wtATTR extracts, KEN hATTR extracts, SHI ALλ liver extracts, and TAL ALκ liver extracts. The log-transformed EC 50 , E.C. 50The maximum levels of binding in the assay are shown. Conditions marked "na" were not tested. As shown in Table E4, humanized anti-amyloid antibodies fused to p5 or p5R were able to bind to a variety of amyloid fibrils and amyloid extracts. VH6 / VL3-p5, VH9 / VL4-p5, and VH9 / VL4-p5R bound all fibrils and extracts tested with higher affinity than m11-1F4 and all other control antibodies (EC 50 VH9 / VL4-p5R generally has a lower EC than VH9 / VL4-p5. 50 VH9 / VL4-p5 generally showed lower EC than VH6 / VL3-p5. 50 showed. [Table 12-1] [Table 12-2]

[0321] Example 4. Wil fibril substrate pulled down by humanized anti-amyloid antibody The ability of anti-amyloid antibodies to pull down substrate was tested as described in PCT / US2020 / 060596. mIgG-p5 provided superior binding to Wil fibrils and amyloid extracts in the pull-down assay (Figure 6). The ability of VH9 / VL4 parent and variants to pull down substrate was significantly reduced compared to mIgp5, as shown in Table E5 below. In Table E5, values ​​shown are percent binding and cells without data represent antibody / substrate combinations that were not tested. [Table 13]

[0322] The ability of humanized anti-amyloid antibodies to act as opsonins for amyloid fibrils (i.e., promote phagocytosis of amyloid fibrils) was tested as described in PCT / US2020 / 060596. VH9 / VL4-p5 and VH9 / VL4-p5R promoted rVλ6Wil fibril uptake better than VH6 / VL3-p5 and VH6 / VL3-p5R, consistent with the differences in the ELISA binding data described above (see Figures 7A and 7B). As shown in Figure 7C, VH9 / VL4 without peptide was roughly as good as VH6 / VL3 bound with p5 or p5R, and many times better than VH6 / Vl3 without peptide. VH9 / VL4 alone was a better opsonin than c11-1F4 (Figure 7B). VH6 / VL3-p5 and VH6 / VL3-p5R promoted fibril uptake at levels comparable to mIgp5 and performed better than c11-1F4 (Figure 7A). VH9 / VL4-p5 and VH9 / VL4-p5R also promoted fibril uptake at levels comparable to mIgp5 and performed better than c11-1F4 (Figure 7B). Surprisingly, humanized anti-amyloid antibodies conjugated to the p5 or p5R peptides provided significantly better opsonization than c11-1F4.

[0323] Example 5. Fed-batch and perfusion production of VH9-D54E / VL4-N33S-p5R; amyloid substrate binding Fed-batch and other semi-continuous culture methods are often used for recombinant protein and antibody production due to their high culture density, high product yield, and flexible productivity applications. However, high density cultures can affect the activity of inhibitory enzymes and toxins that can be detrimental to the overall production yield or strategy. Evaluation of early antibody-peptide fusion protein construct expression by mammalian cells showed that the fusion protein was highly susceptible to cleavage, likely by host cell derived proteases. Perfusion is a type of continuous culture method that avoids the problems associated with high density culture methods (e.g., fed-batch), but perfusion suffers from lower production yields and increased complexity. In this example, VH9-D54E / VL4-N33S-p5R (with VSPSV spacer) was isolated and assessed for purity (percent intact fusion protein) using the exemplary fed-batch and perfusion culture methods outlined in Figure 8A. The binding affinity of the generated antibody-peptide fusion proteins was tested using fed-batch and perfusion culture methods, with the substrate being the amyloid-like fibril rVλ6WIL (FIG. 8B).

[0324] Chinese Hamster Ovary cells were used for protein production by fed-batch and perfusion culture methods. Cells were cultured at 0.1–2x10 with fresh medium (chemically defined, protein-free) while spinning at 120 RPM. 6 The cells were seeded at a density of 100x10 cells / mL in 3-15 cm plates and incubated at 37°C (5% CO2). For the generation of VH9-D54E / VL4-N33S-VSPSV-p5R, 50-200x10 cells were transfected the day before with a plasmid encoding the antibody heavy chain containing VH9-D54E and another plasmid encoding the antibody light chain containing VL4-N33S-VSPSV-p5R. 6 1x10 cells 6The cells were passaged in fresh medium at 1000 cells / mL. The cells were washed and transferred to fresh medium for either fed-batch or perfusion culture. The secreted antibody-peptide fusion protein (VH9-D54E / VL4-N33S-VSPSV-p5R) was harvested from the fed-batch culture after about 7 days. The harvested antibody-peptide fusion protein was then applied to a Protein A chromatography column followed by an anion exchange chromatography column. The eluate from the anion exchange column was collected and the isolated antibody-peptide fusion protein from this fed-batch culture method was designated FB.1 (Fed-batch 1).

[0325] Table E6 shows the mass spectrometry purity analysis of the antibody-peptide fusion proteins using the alternative fed-batch method. [Table 14]

[0326] To remedy the disconnection observed during fed-batch cell culture, perfusion cell culture was evaluated. Perfused cells were grown in a bioreactor where the cell culture was passed through a membrane cartridge that retained the cells but allowed the protein to pass away from the cells and cell debris. Secreted antibody-peptide fusion protein (VH9-D54E / VL4-N33S-VSPSV-p5R) was collected after 1 or 7 days and immediately transferred to sample processing. The secreted antibody-peptide fusion protein was then applied to a Protein A chromatography column followed by an anion exchange chromatography column. The eluate from the anion exchange column was collected and further subjected to cation exchange (CEX) chromatography where the sample was loaded at a loading density of 20.0-50.0 g / L resin, washed with a wash buffer (pH 5.5) containing 260 mM NaCl, and then eluted with an elution buffer (pH 5.5) containing 400 mM NaCl. Application of the antibody-peptide fusion protein to a CEX column allowed the separation of the intact antibody-peptide fusion protein from the cleaved forms of the antibody-peptide fusion protein. The cleaved forms of the antibody-peptide fusion protein were removed with the wash buffer during the washing steps. The intact antibody-peptide fusion protein was eluted with the elution buffer. The eluate from the CEX column was collected and analyzed by sodium dodecyl sulfate capillary electrophoresis (CE-SDS) to determine the purity of the isolated antibody-peptide fusion protein. Purity of >99% was achieved using the perfusion method with CEX column chromatography (Table E7). This eluate was designated PF.1 (perfusion 1) and was prepared for amyloid substrate binding analysis. [Table 15]

[0327] To assess the functional consequences of antibody-peptide fusion protein purity, obtained via fed-batch or perfusion mode production schemes, FB.1 and PF.1 antibody-peptide fusion proteins were prepared for amyloid substrate binding analysis. Amyloid-like fibril rVλ6WIL was used as a substrate to investigate the binding affinity of FB.1 or PF.1. FB.1 antibody-peptide fusion protein was added to wells in two-fold serial dilutions starting at 100 nM, and PF.1 antibody-peptide fusion protein was added to wells in two-fold serial dilutions starting at 5 nM. Detection of bound FB.1 or PF.1 was assessed by measuring time-resolved fluorescence after addition of a biotinylated goat anti-human Fc-reactive secondary antibody and a streptavidin-europium conjugate. The mean and standard deviation (SD) of three replicates were calculated, and potency (EC50) was determined after fitting with a sigmoidal four-parameter logistic (4PL) equation (Prism) with a logarithmic x-axis (Figure 8B).

[0328] The estimated potency (EC50) value for binding of FB.1 to rVλ6WIL was 13.4 nM, whereas the EC50 value for binding of PF.1 to rVλ6WIL was 0.15 nM. These data demonstrate that the highly pure intact antibody-peptide fusion protein (PF.1) produced using perfusion culture and cation exchange chromatography methods has a significantly higher binding affinity for rVλ6WIL amyloid substrate than the antibody-peptide fusion protein (FB.1) produced using fed-batch culture methods. For all subsequent examples, the antibody-peptide fusion proteins were produced using the perfusion culture and cation exchange chromatography methods described in this example.

[0329] Example 6. Biodistribution of PF.1 (VH9-D54E / VL4-N33S-p5R) in mice For this study, PF.1 (VH9-D54E / VL4-N33S-p5R with VSPSV spacer) was expressed by stably transfected CHO cells and produced by perfusion tissue culture for 1 day. PF.1 antibody-peptide fusions were purified by protein A and cation exchange chromatography as described in Example 5. PF.1 antibody-peptide fusions and control hIgG1 were radiolabeled with iodine-125 by oxidative incorporation into tyrosine side chains. Free radioactive iodine was separated by size-exclusion chromatography and radioactive purity was assessed by SDS-PAGE and autoradiography (Figure 9A). Both heavy and light chain proteins of the reagents were radiolabeled with no evidence of free radioactive iodine. The light chain of PF.1 was homogeneous (single species) and had a clearly higher molecular weight compared to the control IgG1 due to the presence of the p5R peptide.

[0330] Approximately 100 μCi (10 μg of reagent) was injected intravenously (IV) into the lateral tail vein of mice with systemic AA amyloidosis involving mainly the liver and spleen. 24 hours after injection, mice (n=3 per group) were euthanized by isoflurane overdose and SPECT / CT images were acquired ( FIG. 9B ). 125I-PF.1 images revealed reduced radioactivity in the liver, spleen and GI tract of AA mice, which contrasted with 125I-hIgG1 images in AA mice 24 hours after injection. There was also clearly more radioactivity in the atrial region of the mouse heart in animals administered 125I-PF.1 compared to the control reagent, which may indicate higher blood pool radioactivity and scarce cardiac amyloid deposits in this mouse model.

[0331] Immediately thereafter, samples or organs and blood were collected for the measurement of tissue-associated radioactivity as an index of the biodistribution of the reagent in the organs, where biodistribution was defined as the percentage of the injected dose measured per gram of tissue. Three mice per group were tested and the mean biodistribution was calculated with standard deviation (SD). Organ analysis included muscle, liver, pancreas, spleen, left kidney, right kidney, stomach, upper intestine, lower intestine, heart, lungs, and blood. This analysis revealed significantly higher retention of 125I-PF.1 compared to control hIgG1 in liver, spleen, kidney, and stomach (Figure 9C). This suggests that the PF.1 reagent is selectively retained by amyloid in these organs.

[0332] The distribution of the radiolabeled reagent in mouse organs was then evaluated using microautoradiography. Briefly, tissue samples were fixed in 10% buffered formalin for 24 hours, embedded in paraffin blocks, and 6 μm thick sections of tissue were prepared on glass slides. Slides were exposed to photographic emulsion for 3 days and then counterstained with hematoxylin and eosin (H&E) stain. The presence of radioactivity in tissues was revealed by the deposition of black silver grains. In all organs evaluated, binding of 125I-PF.1 was observed in association with amyloid deposits in tissues, with representative sections of liver, spleen, and heart showing specific binding to the pathology (Figure 9D) (bar = 500 μm). In contrast, no retention of the 125I-hIgG1 control was observed.

[0333] Example 7. Binding of PF.1 to amyloid in human tissues Formalin-fixed paraffin-embedded sections were prepared from several human tissues containing ATTR (FIG. 10A), AL (FIGS. 10B and 10C), or ALETC2 (FIG. 10D) amyloid. An additional sample of brain tissue from a patient with Alzheimer's disease was also evaluated (FIG. 10E). Tissues were stained with biotinylated PF.1 with a VSPSV spacer (VH9-D54E / VL4-N33S-p5R; 2 μg / mL in PBS) using standard immunohistochemistry methods and visualized after addition of diaminobenzidine (black arrow). The presence of amyloid in slides from the same tissues was visualized by Congo red fluorescence after staining of the tissue with an alkaline Congo red solution (white arrow).

[0334] PF.1 specifically bound to cardiac amyloid deposits around cardiomyocytes in two samples of ATTRv amyloidosis (T60A KEN and T60A SNO) (FIG. 10A) and two samples of AL amyloidosis (ALκTAL and ALκBAB) (FIG. 10B). Similarly, specific binding to amyloid was observed in AL amyloid deposits in the kidney (ALκHUN and ALκJON) (FIG. 10C). Specific binding of PF.1 to ALECT2 amyloid was observed in kidney and spleen tissue samples (FIG. 10D). Finally, diffuse and core plaques composed of Aβ amyloid in the brains of patients with Alzheimer's disease were positively stained with PF.1 (FIG. 10E).

[0335] These data demonstrate specific reactivity of PF.1 with various types of tissue amyloid deposits in diverse tissues. Thus, pan-amyloid reactivity of PF.1 mediated by the p5R peptide is evident by immunohistochemical staining using tissues from the four most common forms of amyloid-related disease.

[0336] Example 8. Phagocytosis of human AL amyloid extract by PF.1 in vivo Samples of human AL (BAL) amyloid extract were labeled with the pH-sensitive dye succinimidyl-pHrodo Red fluorophore. pHrodo Red exhibits weak fluorescence at pH 7.5 but enhances as the pH becomes more acidic, as seen in phagolysosomes of macrophages after phagocytosis. Human AL (BAL) amyloid extract (approximately 10% w / w pHrodo Red labeled material) was preincubated with PF.1 (VH9-D54E / VL4-N33S-p5R with VSPSV spacer; 500 μg PF.1 per 2 mg AL extract) for approximately 30 min. As a control, AL extract was incubated in a similar volume of PBS without PF.1. Immunodeficient (NU / NU) mice (n=4 per group) were administered 2 mg of PF.1-coated AL extract subcutaneously on the back. Fluorescence emission from pHrodo Red-labeled amyloid was detected by optical imaging of mice under isoflurane (1-2% in air) anesthesia. Mice were imaged 1, 5, 7, 10, and 12 days after amyloid injection...

Claims

1. An antibody-peptide fusion protein comprising: i) a heavy chain and ii) an antibody comprising an amyloid-reactive peptide fused via a linker to the C-terminus of a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 91, and the light chain has an amyloid-reactive peptide fused to the C-terminus of the light chain via a linker and comprises the amino acid sequence of SEQ ID NO: 89, said antibody-peptide fusion protein.

2. The antibody-peptide fusion protein according to claim 1, which binds to human amyloid fibrils.

3. The antibody-peptide fusion protein according to claim 1, which exhibits an EC50 of less than 1.5 nM for an amyloid substrate.

4. The antibody-peptide fusion protein according to claim 1, which is conjugated to a detectable label, said detectable label comprising a fluorescent label or a radioactive label.

5. The antibody-peptide fusion protein according to claim 4, wherein the radioactive label is I-123, I-124, F-18, Zr-89, or Tc-99m.

6. The antibody-peptide fusion protein according to claim 1, which exhibits one or more in vivo characteristics selected from improved in vivo distribution, pan-amyloid reactivity, and enhanced phagocytosis as compared to a reference IgG antibody.

7. The antibody-peptide fusion protein according to claim 1, which binds to rVλ6WIL, Aβ, Aβ(1-40), I AAP, ALκ4, ALλ1, ATTR, α-synuclein, or tau fibrils.

8. A pharmaceutical composition comprising the antibody-peptide fusion protein according to any one of claims 1 to 7 and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to claim 8, which is formulated for intravenous or subcutaneous administration.

10. A polynucleotide encoding the antibody-peptide fusion protein according to any one of claims 1 to 7.

11. A vector comprising the polynucleotide according to claim 10.

12. A host cell comprising the vector according to claim 11.

13. The host cell according to claim 12, which is a mammalian cell, optionally a Chinese hamster ovary (CHO) cell.

14. A method for producing an antibody-peptide fusion protein, comprising: a) culturing a host cell containing a vector expressing the antibody-peptide fusion protein according to any one of claims 1 to 7 under perfusion cell culture conditions suitable for the expression of the antibody-peptide fusion protein; and b) recovering the antibody-peptide fusion protein every about 12 to 36 hours The method as described above.

15. The method according to claim 14, further comprising applying the antibody-peptide fusion recovered in step b) to a cation exchange chromatography column and eluting the antibody-peptide fusion protein from the cation exchange chromatography column.

16. The method according to claim 15, wherein the antibody-peptide fusion protein is eluted separately from the cleaved antibody-peptide fusion protein.

17. The method according to claim 14, wherein the host cell is a CHO cell.

18. The method according to claim 14, further comprising determining the purity of the antibody-peptide fusion protein, wherein the purity of the antibody-peptide fusion protein is determined using one or more analytical methods including sodium dodecyl sulfate capillary electrophoresis (CE-SDS), liquid chromatography (LC), mass spectrometry (MS), or combinations thereof.

19. The method according to claim 14, wherein the antibody-peptide fusion protein is purified to at least 90% intact antibody-peptide fusion protein.

20. A therapeutically effective amount of the antibody-peptide fusion protein according to claim 1 for use in a method of treating a subject having an amyloid-related disorder comprising amyloid deposits.

21. A therapeutically effective amount of the pharmaceutical composition according to claim 8 for use in a method of treating a subject having an amyloid-related disorder comprising amyloid deposits.

22. The antibody-peptide fusion protein or pharmaceutical composition for use according to claim 20 or 21, wherein the amyloid-related disorder is systemic or localized amyloidosis.

23. The amyloid-related disorder is an antibody-peptide fusion protein or pharmaceutical composition for use according to claim 20 or 21, selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRw, AA, AApAI, AApAII, AGel, ALys, ALECT2, AFib, ACys, ACal, AMed, AβAPP, APro, AIns, APrP, Parkinson's disease, Alzheimer's disease, and Aβ amyloidosis.

24. The amyloid-related disorder is systemic amyloidosis, and the antibody-peptide fusion protein or pharmaceutical composition for use according to claim 20 or 21.

25. The amyloid-related disease is selected from the group consisting of AL, AH, Aβ2M, ATTRv, ATTRwt, AA, AApAI, AApAII, AGel, ALys, ALECT2, AFib, and ACys, and optionally, the amyloid-related disease is AL or ATTR. The antibody-peptide fusion protein or pharmaceutical composition for use according to claim 24.

26. The amyloid deposit is opsonized by the antibody-peptide fusion protein, and the antibody-peptide fusion protein or pharmaceutical composition for use according to claim 20 or 21.

27. Treating the subject with the antibody-peptide fusion protein causes phagocytosis of the amyloid deposit, and the antibody-peptide fusion protein or pharmaceutical composition for use according to claim 20 or 21.

28. The antibody-peptide fusion protein according to claim 1 for use in a method of treating a subject having or suspected of having an amyloid-based disease, a) the subject is i) administering the antibody-peptide fusion protein to the subject, the antibody-peptide fusion protein comprising a detectable label, the administering, and ii) determining whether a signal associated with the detectable label can be detected from the subject to determine whether there is an amyloid deposit, and b) applying an amyloidosis treatment to the subject if the signal is detected comprising the antibody-peptide fusion protein according to claim 1.

29. The pharmaceutical composition according to claim 8 for use in a method of treating a subject having or suspected of having an amyloid-based disease, a) wherein the subject is, i) administering the antibody-peptide fusion protein to the subject, the antibody-peptide fusion protein comprising a detectable label, said administering, and ii) determining whether a signal associated with the detectable label can be detected from the subject to determine whether the subject has amyloid deposits, and b) when the signal is detected, applying an amyloidosis treatment to the subject The pharmaceutical composition according to claim 8, comprising.

30. An antibody-peptide fusion protein for use according to claim 28 or 29, wherein when no signal is detected, the subject is monitored for subsequent growth of amyloid deposits or a pharmaceutical composition.

31. The antibody-peptide fusion protein or pharmaceutical composition for use according to claim 28 or 29, further comprising determining the intensity of the signal and comparing the signal to a threshold value, and when the threshold value is exceeded, the subject is determined to have amyloid deposits

32. The antibody-peptide fusion protein or pharmaceutical composition for use according to claim 28 or 29, wherein the antibody-peptide fusion protein is detected by SPECT / CT imaging, PET / CT imaging, gamma scintigraphy, or optical imaging

33. The antibody-peptide fusion protein or pharmaceutical composition for use according to claim 28 or 29, wherein the amyloidosis treatment comprises administering to the subject the antibody-peptide fusion protein according to any one of claims 1 to 7

34. The pharmaceutical composition for use according to claim 29, wherein the amyloidosis treatment comprises administering to the subject the pharmaceutical composition according to claim 8

35. A method of identifying amyloid deposits in a subject, comprising administering the antibody-peptide fusion protein according to claim 1, the antibody-peptide fusion protein comprising a detectable label, said administering, and detecting a signal from the antibody-peptide fusion protein

36. A method for identifying amyloid deposits in a subject, comprising administering the pharmaceutical composition according to claim 8, wherein the antibody-peptide fusion protein comprises a detectable label, and said administering and detecting a signal from the antibody peptide fusion protein.

37. A method for monitoring amyloid clearance in a subject, comprising: contacting an amyloid substrate in the subject with the antibody-peptide fusion protein according to claim 1, wherein the antibody-peptide fusion protein comprises a detectable label, and the peptide of the antibody-peptide fusion protein has a binding affinity for the amyloid substrate, said contacting, and determining a signal from the detectable label, thereby detecting the amyloid clearance comprising the method.

38. A method for monitoring amyloid clearance in a subject comprising the pharmaceutical composition according to claim 8, comprising: contacting an amyloid substrate in the subject with the pharmaceutical composition according to claim 8, wherein the antibody-peptide fusion protein comprises a detectable label, and the peptide of the antibody-peptide fusion protein has a binding affinity for the amyloid substrate, said contacting, and determining a signal from the detectable label, thereby detecting the amyloid clearance comprising the method.

39. The subject is a human, the antibody-peptide fusion protein according to any one of claims 1 to 7, the use according to claim 20 or 21, or any one of claims 35 to 38 the method described in the item.

40. A kit comprising the antibody-peptide fusion protein according to any one of claims 1 to 7.

41. A kit comprising the pharmaceutical composition according to claim 8.

42. An antibody-peptide fusion protein, wherein the fusion protein comprises a first polypeptide, a second polypeptide, a third polypeptide, and a fourth polypeptide, the first polypeptide and the second polypeptide each comprise the amino acid sequence shown in SEQ ID NO: 89, and the third polypeptide and the fourth polypeptide each comprise the amino acid sequence shown in SEQ ID NO: 91, said antibody-peptide fusion protein.

43. A therapeutically effective amount of an antibody-peptide fusion protein for use in a method of treating AL in a subject, wherein the antibody-peptide fusion protein comprises: i) a heavy chain and ii) an antibody comprising an amyloid-reactive peptide fused via a linker to the C-terminus of the light chain, wherein the heavy chain comprises the amino acids shown in SEQ ID NO: 91 and the amyloid-reactive peptide fused via a linker to the C-terminus of the light chain and light chain comprises the amino acid sequence shown in SEQ ID NO: 89, said antibody-peptide fusion protein. **Claim 44** A therapeutically effective amount of an antibody-peptide fusion protein for use in a method of treating ATTR in a subject, wherein the antibody-peptide fusion protein comprises: i) a heavy chain and ii) an antibody comprising an amyloid-reactive peptide fused via a linker to the C-terminus of the light chain, wherein the heavy chain comprises the amino acids shown in SEQ ID NO: 91 and the amyloid-reactive peptide fused via a linker to the C-terminus of the light chain and light chain comprises the amino acid sequence shown in SEQ ID NO: 89, said antibody-peptide fusion protein.

Images