Modified immunoglobulins for targeting amyloid deposits

Abstract

To provide modified immunoglobulins for targeting amyloid deposits.SOLUTION: Provided herein are modified immunoglobulins comprising an amyloid reactive peptide joined to an antibody, as well as humanized antibodies that bind to human amyloid fibrils and antibody-peptide fusion proteins. Also provided herein are methods of treating amyloid-based diseases by administering a modified immunoglobulin, humanized antibody, or antibody-peptide fusion protein.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Application No. 62 / 936,002, filed November 15, 2019, and U.S. Provisional Application No. 63 / 074,912, filed September 4, 2020, the contents of which are incorporated herein by reference in their entirety.

[0002] Submission of sequence listings in ASCII text files. The contents of the following submission in ASCII text file are incorporated herein by reference in their entirety: a computer-readable format (CRF) sequence listing (filename: 165992000140SEQLIST.TXT, date: November 12, 2020, size: 52KB).

[0003] This application relates to modified immunoglobulins for targeting amyloid deposits, humanized antibodies that bind to human amyloid fibrils, antibody-peptide fusion proteins, and methods for using them. [Background technology]

[0004] Amyloidosis is a fatal protein folding disorder characterized by the aggregation and deposition of proteinifibrillars and heparan sulfate proteoglycans in vital organs and tissues (Non-Patent Documents 1, 2, 3, and 4). The continuous accumulation of amyloid inevitably leads to organ dysfunction and severe morbidity or death. Deposits can be cerebral, as in patients with Alzheimer's disease, Huntington's disease, or prion diseases, or peripheral, as in patients with light chain (AL) amyloidosis and type 2 diabetes. Further subgrouping into focal and systemic amyloidosis indicates whether precursor proteins are produced locally (at the deposition site) or circulate in the bloodstream and deposited at distant anatomical sites (Non-Patent Document 5). While amyloid can affect any organ or tissue, in patients with familial or sporadic peripheral amyloid disease, the kidneys, pancreas, liver, spleen, nerve tissue, and heart constitute the primary deposition sites. Alzheimer's disease currently affects more than 4 million Americans, and this number is estimated to increase to over 16 million by 2050. This is the most common form of amyloidosis and has the greatest socioeconomic impact. In contrast, peripheral (or systemic) amyloidosis is an orphan disorder, but it accounts for more than 5,000 new cases per year in the United States alone.

[0005] Of these, the most common peripheral amyloidosis is light chain-related (AL) amyloidosis, a sporadic monoclonal plasmacytosis resulting from the deposition of fibrils composed of immunoglobulin light chain proteins. AL accounts for approximately two-thirds of all peripheral amyloid cases and has an estimated incidence of about 1.4 per 100,000 people per year in the United States, comparable to acute lymphoblastic leukemia and chronic myeloid leukemia (Non-Patent Literature 6). While AL is common, accounting for one-fifth of related plasmacytosis multiple myelomas, it can be said to be more destructive, partly due to its rapidly progressing organ destruction, lack of effective anti-amyloid drugs, and the inability to effectively diagnose the disease before organ failure occurs, resulting in a median survival time of only 13.2 months. Fewer than 5% of all AL patients survive for more than 10 years from diagnosis (Non-Patent Literature 7). Furthermore, the median survival time for patients with cardiac AL amyloidosis is less than 5 months.

[0006] ATTR is a form of systemic amyloidosis. 25% of patients die within 24 months of diagnosis (Non-Patent Literature 8). Current therapies cannot prevent organ damage. ATTR amyloidosis is caused by transthyretin (TTR) fibrils. Transthyretin is a protein produced by the liver that helps transport thyroid hormones and vitamin A in the blood. Normally, TTR is a tetramer consisting of four single-chain monomers. In hereditary ATTR amyloidosis, it is thought that mutations in the TTR gene destabilize the protein, causing the tetramer to dissociate into monomers, which then aggregate into amyloid fibrils. In wild-type ATTR amyloidosis, normal TTR proteins become unstable, misfold, and form amyloid fibrils.

[0007] These amyloid fibrils then accumulate in multiple organs throughout the body. For example, in the wrist, they accumulate in a narrow passage called the carpal tunnel. This can cause carpal tunnel syndrome, which can lead to numbness and tingling in the hand and arm. In the spinal canal, it can cause narrowing of the spine (spinal stenosis). In the heart, it can cause heart failure and / or an irregular heartbeat called atrial fibrillation.

[0008] Another common 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 regions of Europe, and its frequency varies among ethnic groups (Non-Patent Literature 9). In areas where familial Mediterranean fever is prevalent and untreated, the incidence of AA can be 100%. In Europe, the incidence of AA is estimated at 0.86% based on an autopsy study conducted in Denmark (Non-Patent Literature 10), but in patients with rheumatoid or psoriatic arthritis, the incidence of AA can be as high as 26%. Such high prevalence may necessitate screening programs to detect the disease earlier. Amyloid deposition is associated with a persistent increase in plasma concentrations of serum amyloid protein A (sAA), a precursor of amyloid fibrils (Non-Patent Literature 11). Although AA differs from AL in the type of precursor protein it deposits, both share common mechanistic features related to fibril formation and deposition (Non-Patent Documents 12, 13).

[0009] In addition to disorders for which the pathogenesis of amyloid is well established, fibrous deposits with the structural and coloring properties of amyloid have been identified in other syndromes, but their association with those disease states has not yet been established. For example, in type 2 diabetes, islet amyloid precursor protein (IAPP) is deposited as amyloid in the islets of Langerhans (Non-Patent Literature 14). Aggregation of IAPP leads to oligomeric structures that are toxic to pancreatic cells (Non-Patent Literature 15). Therefore, it has been suggested that the formation of IAPP amyloid in patients with type 1 diabetes contributes to the destruction of β-cells and leads to insulin-dependent transition (Non-Patent Literature 16). In another example, plaques containing amyloid fibrils composed of apolipoprotein AI have been identified in more than half of patients with atherosclerotic carotid arteries (Non-Patent Literature 17, Non-Patent Literature 18). Although the deposition of these fibrils was more common in older patients, apoA-I is undoubtedly present in the early stages of plaque formation (Non-Patent Literature 19). As a final example, Apo-AI amyloid has recently been identified in the knee meniscus of patients who have undergone knee replacement surgery and may contribute to the physical deterioration of the joint (Non-Patent Literature 20).

[0010] In total, more than 29 proteins have been chemically or serologically identified as components of fibrils in amyloid deposits. The properties of these proteins are crucial for differentiating diseases, determining treatments, and establishing prognosis. Amyloid fibrils are associated with a clinically heterogeneous group of diseases and can be formed from structurally different and functionally diverse precursor proteins, but the deposits themselves share several remarkably similar features, including fibril structure, fibril epitopes, and the development of similar accessory molecules, including heparan sulfate proteoglycans (HSPGs). Amyloid is a heterogeneous complex that, in addition to profibrils, contains glycosaminoglycans (GAGs), particularly perlecan HSPG (Non-Patent Literature 21, 22, 23, 24, 25, and 26). Figure 1 shows a partial list of amyloid and amyloid-related disorders.

[0011] To date, the most effective therapeutic intervention to remove amyloid deposits that can promote organ function recovery and improve prognosis has involved the use of amyloid-reactive antibodies as a means of immunotherapy. Several immunotherapies (antibodies) have been developed for amyloid-related diseases, including the 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. Each of these approaches has limitations or has not met the primary outcomes in late-stage clinical trials (phase 2 / 3). [Prior art documents] [Non-patent literature]

[0012] [Non-Patent Document 1] Merlini, G. et al. (2003) N. Engl. J. Med. 349, 583-596 [Non-Patent Document 2] Merlini, G. et al. (2004) J. Intern. Med. 255, 159-178 [Non-Patent Document 3] De Lorenzi, E. et al. (2004) Curr. Med. Chem. 11, 1065-1084 [Non-Patent Document 4] Merlini, G. (2004) Neth. J. Med. 62, 104-105 [Non-Patent Document 5] Westermark, P. et al. (2007) Amyloid.14, 179-183 [Non-Patent Document 6] 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

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Non-licensed literature 9

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[0014] Methods for detecting and treating amyloidosis, or methods using modified immunoglobulins provided herein, are also provided herein.

[0015] Furthermore, nucleic acids encoding modified immunoglobulins are provided herein. In some embodiments, host cells containing nucleic acids encoding modified immunoglobulins are provided herein. In some embodiments, the host cells are CHO cells.

[0016] In one embodiment, the present invention provides a modified immunoglobulin comprising an amyloid-reactive peptide and an Ig antibody or a functional fragment thereof that binds to human amyloid fibrils, and the Ig antibody or The functional fragment comprises a heavy chain and a light chain, and the peptide and the Ig antibody or its functional fragment are linked together at the N-terminus of the Ig light chain and / or the N-terminus and / or C-terminus of the Ig heavy chain.

[0017] In some embodiments, the amyloid-reactive peptide comprises an amino acid sequence having at least 85% sequence identity with any one of the amino acid sequences described in SEQ ID NOs: 1 to 14.

[0018] In some embodiments, the amyloid-reactive peptide and the Ig antibody or its functional fragment are linked together at the N-terminus of the Ig light chain.

[0019] In some embodiments, the modified immunoglobulin includes a spacer sequence between the amyloid-reactive peptide and the Ig antibody or its functional fragment.

[0020] In some embodiments, the modified immunoglobulin comprises at least two amyloid-reactive peptides, which are the same peptide or different peptides.

[0021] In some embodiments, the Ig antibody or its functional fragment comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDRL1 as described in SEQ ID NO: 20, CDRL2 as described in SEQ ID NO: 21, and CDRL3 as described in SEQ ID NO: 22, and the VH includes CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, and CDRH3 as described in SEQ ID NO: 19.

[0022] In some embodiments, the Ig antibody or its functional fragment is a chimeric antibody or its functional fragment.

[0023] In some embodiments, the Ig antibody or its functional fragment comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19, or b) the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 20, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19.

[0024] In some embodiments, the Ig antibody or its functional fragment includes a human framework sequence.

[0025] In some embodiments, the Ig antibody includes a human Fc region.

[0026] In some embodiments, the modified immunoglobulin comprises at least two amyloid-reactive peptides, the peptides being the same peptide or different peptides.

[0027] In some embodiments, the modified immunoglobulin binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, Alλ1, or ATTR fibrils.

[0028] In another embodiment, it binds to human amyloid fibrils fused with amyloid-reactive peptides. An antibody-peptide fusion protein containing an antibody is provided herein, wherein the antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH).

[0029] In some embodiments, VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.

[0030] In some embodiments, the antibody is a chimeric antibody.

[0031] In some embodiments, a) VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19, or b) VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 20, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22 VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequences described in SEQ ID NOs: 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19, or c) VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs: 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequences described in SEQ ID NOs: 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.

[0032] In some embodiments, VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 64, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 73, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.

[0033] In some embodiments, VL includes Leu at position 46 and Phe at position 87, and VH includes Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94.

[0034] In some embodiments, VL includes the amino acid sequence described in SEQ ID NO: 36, and VH includes the amino acid sequence described in SEQ ID NO: 55.

[0035] In some embodiments, VL comprises one or more amino acid residues selected from the group consisting of: Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87. In some embodiments, VH comprises one or more amino acid residues selected from the group consisting of: Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Val at position 93, and Thr at position 94 (amino acid positions are numbered according to the Kabat numbering system).

[0036] In some embodiments, VL includes Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH includes Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, and T at position 89. Includes hr, Val at position 93, and Thr at position 94.

[0037] In some embodiments, VL includes Leu at position 46 and Phe at position 87, and VH includes Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94.

[0038] In some embodiments, VL includes the amino acid sequence described in the group consisting of SEQ ID NOs. 32 to 42.

[0039] In some embodiments, VH includes the amino acid sequence described in the group consisting of SEQ ID NOs: 43 to 63.

[0040] In some embodiments, VL includes the amino acid sequence described in SEQ ID NO: 34, and VH includes the amino acid sequence described in SEQ ID NO: 48.

[0041] In some embodiments, VL includes the amino acid sequence described in SEQ ID NO: 35, and VH includes the amino acid sequence described in SEQ ID NO: 51.

[0042] In some embodiments, the amyloid-reactive peptide comprises the amino acid sequences described in SEQ ID NOs: 1 to 14.

[0043] In some embodiments, the amyloid-reactive peptide is fused to the N-terminus of VL or VH.

[0044] In some embodiments, an amyloid-reactive peptide is fused to the N-terminus of VL or VH by a spacer. In some embodiments, the spacer is a peptide spacer. In some embodiments, the spacer contains the amino acid sequence GGGYS.

[0045] In some embodiments, the antibody-peptide fusion protein binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, Alλ1, or ATTR fibrils.

[0046] In another embodiment, a pharmaceutical composition comprising a modified immunoglobulin or an antibody-peptide fusion protein is provided herein.

[0047] In another embodiment, nucleic acids (or more) encoding modified immunoglobulins and antibody-peptide fusion proteins are provided herein. In another embodiment, vectors comprising nucleic acids (or more) are provided herein. In another embodiment, host cells comprising vectors are provided herein.

[0048] In another aspect, the present invention provides a method for producing a modified immunoglobulin or antibody-peptide fusion protein, comprising culturing the host cells described in paragraph

[0045] under conditions suitable for the expression of a vector encoding the modified immunoglobulin or antibody-peptide fusion protein, and recovering the modified immunoglobulin or antibody-peptide fusion protein.

[0049] In another aspect, the present invention provides a method for treating a subject having amyloid-related disorder, comprising administering to the subject an effective amount of modified immunoglobulin or antibody-peptide fusion protein.

[0050] In some embodiments, amyloid-related disorders are amyloidosis.

[0051] In some embodiments, amyloid-related disorders are selected from the group consisting of AL, AH, Aβ2M, ATTR, transthyretin, AA, AApoAI, AApoAII, AGel, Alys, ALEct2, AFib, ACYs, ACa, AMed, AIAPP, APro, AIns, APrP, or Aβ amyloidosis.

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

[0053] In another embodiment, the present invention provides a method for targeting amyloid deposits for clearance, comprising contacting the amyloid deposits with a modified immunoglobulin or antibody-peptide fusion protein provided herein. In some embodiments, the amyloid deposits are removed. In some embodiments, the amyloid deposits are opsonized by the modified immunoglobulin or antibody-peptide fusion protein.

[0054] In another embodiment, a method for targeting amyloid deposits for clearance is provided herein, comprising contacting the amyloid deposits with a modified immunoglobulin or an antibody-peptide fusion protein.

[0055] In some embodiments, targeting amyloid deposits for clearance results in the clearance of amyloid deposits.

[0056] In some embodiments, clearance arises from the opsonization of amyloid deposits.

[0057] In some embodiments, the half-life of amyloid-reactive peptides in modified immunoglobulins or antibody-peptide fusion proteins is increased by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or more compared to amyloid-reactive peptides alone.

[0058] In another embodiment, a method for generating a modified immunoglobulin is provided herein, comprising providing a first expression vector and a second expression vector, wherein the first expression vector comprises a first nucleic acid sequence encoding an Ig antibody light chain or a functional fragment thereof, the second expression vector comprises a second nucleic acid sequence encoding an Ig antibody heavy chain or a functional fragment thereof, the first expression vector and / or the second expression vector comprises a third nucleic acid sequence encoding a first peptide, the third nucleic acid sequence being located adjacent to the first nucleic acid sequence and / or the second nucleic acid sequence, and inserting the first expression vector and the second expression vector into a cell, wherein the expression of the first expression vector and the second expression vector in the cell results in an immunoglobulin linked to the first peptide.

[0059] In some embodiments, the first expression vector and / or the second expression vector includes a fourth nucleic acid sequence encoding a second peptide, the fourth nucleic acid sequence being located adjacent to the first and / or second nucleic acid sequence.

[0060] In some embodiments, the expression of the first and second expression vectors in cells results in immunoglobulins linked to the first and second peptides.

[0061] In some embodiments, the spacer nucleic acid sequence is located between the third nucleic acid sequence and the first nucleic acid sequence and / or the second first nucleic acid sequence.

[0062] In another aspect, the present invention relates to subjects suffering from amyloid-related diseases, or amyloid-related diseases The present invention provides a method for treating a subject suspected of having a disease, comprising: i) administering a modified immunoglobulin or antibody-peptide fusion protein to the subject, wherein the modified immunoglobulin or antibody-peptide fusion includes a detectable label; ii) determining whether a signal associated with the detectable label can be detected from the subject; and b) if a signal is detected, administering amyloidosis treatment to the subject.

[0063] In some embodiments, if no signal is detected, the method further includes monitoring the subject for the subsequent development of amyloid deposits.

[0064] In some embodiments, the method further includes determining the intensity of a signal and comparing the signal to a threshold, wherein if the threshold is exceeded, the subject is determined to have amyloid deposits.

[0065] In some embodiments, amyloidosis treatment involves administering a modified immunoglobulin or antibody-peptide fusion protein to the subject.

[0066] In some embodiments, administration of modified immunoglobulins or antibody-peptide fusion proteins results in the clearance of amyloid deposits in the subject.

[0067] In another aspect, the present invention provides a method for identifying amyloid deposits in a subject, comprising administering a modified immunoglobulin or antibody-peptide fusion protein to the subject, wherein the modified immunoglobulin or antibody-peptide fusion protein includes a detectable label, and detecting a signal from the modified immunoglobulin or antibody-peptide fusion protein.

[0068] In some embodiments, subjects are determined to have amyloid-free or unspecified monoclonal immunoglobulinemia (MGUS), multiple myeloma (MM), or one or more related plasma cell disorders.

[0069] In another aspect, the present invention provides a method for detecting a ligand, comprising: detecting a modified immunoglobulin or antibody-peptide fusion as described in any one paragraph, wherein the peptide of the modified immunoglobulin or antibody-peptide fusion protein has binding affinity to the ligand; contacting the ligand with the modified immunoglobulin or antibody-peptide fusion protein; and determining a signal from the detectable label, thereby detecting the ligand.

[0070] In some embodiments, modified immunoglobulins or antibody peptide fusion proteins are conjugated with a detectable label.

[0071] In some embodiments, the modified immunoglobulin or antibody-peptide fusion protein includes a spacer located at the N-terminus of the amyloid-reactive peptide.

[0072] In some embodiments, the amyloid-reactive peptide is ligated to the N-terminus of the VH of the antibody.

[0073] In one embodiment, a humanized antibody that binds to human amyloid fibrils is provided herein, the humanized antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, and SEQ ID NO: 18 The CDR-H2 contains an amino acid sequence, and the CDR-H3 contains an amino acid sequence described in Sequence ID No. 19, wherein VL contains one or more amino acid residues selected from the group consisting of: Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH contains one or more amino acid residues selected from the group consisting of: Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Val at position 93, and Thr at position 94 (where the amino acid positions are numbered according to the Kabat numbering system).

[0074] In some embodiments, VL includes Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH includes Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94.

[0075] In some embodiments, VL includes Leu at position 46 and Phe at position 87, and VH includes Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94.

[0076] In some embodiments, VL includes the amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42.

[0077] In some embodiments, VH includes the amino acid sequence described in the group consisting of SEQ ID NOs: 44 to 63.

[0078] In some embodiments, VL includes the amino acid sequence described in SEQ ID NO: 34, and VH includes the amino acid sequence described in SEQ ID NO: 48.

[0079] In some embodiments, VL includes the amino acid sequence described in SEQ ID NO: 35, and VH includes the amino acid sequence described in SEQ ID NO: 51.

[0080] In another embodiment, a humanized antibody that binds to human amyloid fibrils is provided herein, the humanized antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), where a) VL comprises CDR-L1 containing the amino acid sequences described in SEQ ID NOs. 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22, and VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19, or b) VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 20, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 21 -VH includes CDR-L2 and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequences described in SEQ ID NOs: 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19, or c)VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs: 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequences described in SEQ ID NOs: 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.

[0081] In some embodiments, VL includes one or more amino acid residues selected from the group consisting of: Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH includes one or more amino acid residues selected from the group consisting of: Including: Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Val at position 93, and Thr at position 94 (where the amino acid positions are numbered according to the Kabat numbering system).

[0082] In some embodiments, the antibody is a full-length antibody, a Fab fragment, or an scFv.

[0083] In some embodiments, the antibody includes an Fc region.

[0084] In some embodiments, the Fc region is an Fc region of the IgG1, IgG2, IgG3, or IgG4 isotype.

[0085] In some embodiments, a humanized antibody or antibody-peptide fusion protein is conjugated to a detectable label.

[0086] In some embodiments, the humanized antibody binds to rVλ6Wil fibril, Per125 wtATTR extract, KEN hATTR extract, SHI ALλ liver extract, and / or TAL ALκ liver extract.

[0087] In some embodiments, the humanized antibody binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, Alλ1, or ATTR fibrils.

[0088] In another aspect, the present invention provides a pharmaceutical composition comprising a humanized antibody. In another aspect, the present invention provides nucleic acids (or more) encoding a humanized antibody. In another aspect, a vector comprising nucleic acids (or more) is provided herein. In another aspect, a host cell comprising a vector is provided herein.

[0089] In another aspect, the present invention provides a method for producing a humanized antibody, comprising culturing the host cells described in paragraph

[0088] under conditions suitable for the expression of a vector encoding a humanized antibody, and recovering the humanized antibody.

[0090] In another embodiment, the present invention provides a method for treating a subject having an amyloid-related disorder, comprising administering an effective amount of a humanized antibody to the subject. In some embodiments, the amyloid-related disorder is 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, ACa, AMed, AIAPP, APro, AIns, APrP, or Aβ amyloidosis. In some embodiments, the subject is human.

[0091] In another embodiment, the present invention provides a method for treating a subject who has or is suspected to have an amyloid disorder, the method comprising determining whether the subject has amyloid deposits by detecting a humanized antibody, administering the humanized antibody to the subject, determining whether a signal associated with the detectable label can be detected from the subject, and, if a signal is detected, administering amyloidosis treatment to the subject.

[0092] In some embodiments, if no signal is detected, the method further includes monitoring the subject for the subsequent development of amyloid deposits.

[0093] In some embodiments, this method determines the signal intensity and sets the signal to a threshold value. This further includes comparison, and if a threshold is exceeded, the subject is determined to have amyloid deposits. In some embodiments, amyloidosis treatment includes administering a humanized antibody to the subject.

[0094] In another embodiment, the present invention provides a method for identifying amyloid deposits in a subject, comprising: detecting a humanized antibody; administering the humanized antibody to the subject; and detecting a signal from the humanized antibody.

[0095] In some embodiments, subjects are determined to have amyloid-free or unspecified monoclonal immunoglobulinemia (MGUS), multiple myeloma (MM), or one or more related plasma cell disorders. [Brief explanation of the drawing]

[0096] [Figure 1] A partial list of amyloid and amyloid-related disorders is shown below. [Figure 2] These are schematic diagrams of Ig-peptide fusions. Panel A shows a schematic diagram of peptide p5 (SEQ ID NO: 1) fused to the N-terminus of an Ig light chain. Panel B shows an Ig-peptide fusion, illustrating the Ig structure with two Ig light chains fused to peptide p5 (SEQ ID NO: 1). [Figure 3] A schematic diagram (not to scale) shows the proposed mechanism of action of an Ig-peptide fusion for clearing amyloid deposits. The Ig-peptide binds to amyloid fibrils (or heparan sulfate glycosaminoglycans) through peptide interactions, which recruit macrophages to then engulf (phagocytose) the amyloid to break it down. [Figure 4] The autoradiograph of 125I-Igp5 after SDS-PAGE gel electrophoresis is shown. Under non-reducing conditions, the protein migrated as a single Ig, and when reduced, it was shown to contain the heavy and light chains -p5 and be consistent with an intact Ig molecule. [Figure 5] This shows the in vivo distribution of 125I-Igp5 in AA amyloid mice (AA) and healthy amyloid-free mice (WT) 20 hours after injection. [Figure 6] SPECT / CT images of 125I-Igp5 in AA mice and healthy wild-type controls 20 hours after Igp5 injection are shown, illustrating uptake in the liver and spleen containing amyloid in AA mice, as well as the long blood pool half-life of the reagent in WT animals. H, heart; L, liver; S, spleen. Red / yellow indicates the presence of 125I-Igp5. [Figure 7]This image shows in vitro phagocytosis of pHrodo-red labeled rVλ6Wil fibrils in the presence of human THP1 monocytes / macrophages. Increased fluorescence intensity indicates the presence of labeled amyloid substrate in the low pH environment of macrophage phagolysosomes. MOPC 31c is a mouse monoclonal antibody control that does not resonate with fibrils. [Figure 8] The autoradiograph of 125I-Igp5 after SDS-PAGE gel electrophoresis is shown. Igp5 was purified from tissue culture supernatant, radiolabeled with 125I, and the product was characterized by SDS-PAGE using mouse 11-1F4 (IgG1κ) as a control. The protein was analyzed under both reducing (Red.) and non-reducing (NR) conditions. [Figure 9] The binding of 125I-Igp5 to various amyloid-related substrates is shown. Panel A shows the binding of 125I-m11-1F4 and 125I-Igp5 to κ4-peptide-coated beads or rVλ6Wil fibrils. 125I-m11-1F4 binds to κ4-peptide-coated beads but not to rVλ6Wil fibrils, while 125I-Igp5 binds to both substrates. Panel B shows the quantification of the binding of 125I-Igp5 and m11-1F4 to various synthetic amyloid fibrils and amyloid extracts. Panel C shows the correlation between the binding of 125I-Igp5 and 125I-p5 to their substrates. The binding of 125I-Igp5 to various synthetic amyloid fibrils and amyloid extracts was significantly enhanced compared to m11-1F4, and the reactivity correlated with that of the p5 peptide alone, indicating that the binding is peptide-driven. [Figure 10]The panel shows microautoradiography (ARG) analysis and Congo red staining of various tissues. Panel A shows the retention of 125I-Igp5 in hepatosplenic AA amyloid and amyloid binding in other tissues in mice. ARG and Congo red staining show the specific retention of 125I-Igp5 in hepatosplenic AA amyloid and amyloid binding in other tissues in mice. Panel B shows the results of ARG analysis of amyloid-free tissue. In healthy (WT) mice, no specific reactivity with amyloid-free tissue was observed, and the only identified source of 125I-Igp5 was the blood pool. [Figure 11A] A schematic diagram of mIgG-p5 is shown. From left to right, the region of mIgG-p5 from the C-terminus to the N-terminus is illustrated and includes the Ig light chain sequence ("IgLC", 220 amino acids), spacer sequence, peptide p5 (31 amino acids), spacer sequence, and N-terminal secretory leader sequence. [Figure 11B] The amino acid sequence of the mIgG-p5 region is shown, from left to right, including the N-terminus of the Ig light chain (starting with amino acid residue DVVMTQTP (SEQ ID NO: 82)), the C-terminal spacer sequence of the p5 peptide (amino acid residue VTPTV (SEQ ID NO: 24)), the p5 peptide (amino acid residues KAQKAQAKQAKQAQKAQKAQAKQAKQ (SEQ ID NO: 1)), and the N-terminal spacer sequence (amino acid residues AQAGQAGQAQGGGYS (SEQ ID NO: 23)). The amino acid sequence is shown from the C-terminus to the N-terminus. [Figure 11C] The proposed structural model of peptide p5 fused to the N-terminus of the Ig light chain is shown. [Figure 11D] The autoradiographs of 125I-labeled mIgG-p5, 125I-m11-1F4, and 125I-p5 after SDS-PAGE gel electrophoresis are shown. For each protein, the relative positions of the full-length antibody ("Ig"), heavy chain ("HC"), light chain ("LC"), and peptide p5 are shown under non-reducing ("NR") or reducing ("R") conditions. [Figure 11E]The in vivo distribution of 125I-labeled mIgG-p5 in wild-type amyloid-free mice 24, 48, or 72 hours after injection. The y-axis shows the level of in vivo distribution as a percentage of the injected dose per gram of tissue, and the tissue type is shown on the x-axis. [Figure 11F] This graph shows the in vivo distribution of 125I-labeled mIgG-p5 24, 48, or 72 hours after injection of 125I-mIgG-p5 in mice with AA amyloidosis (primarily affecting the liver and spleen). The y-axis represents the level of in vivo distribution as a percentage of the injected dose per gram of tissue, and the tissue type is indicated on the x-axis. [Figure 11G] Microautography images showing 125I-labeled mIg-p5 in mice with AA amyloidosis 24 hours after injection are shown. 24 hours are shown in black, 48 hours in dark gray, and 72 hours in light gray. [Figure 12] The amino acid sequences of the annotated VH (top, SEQ ID NO: 15) and VL (bottom, SEQ ID NO: 16) of the parent mouse antibody m11-1F4 are shown. The CDR is shown in a box, residues in the canonical framework region are underlined, and residues at the VH-VL interface are shown in bold and italics. [Figure 13A] The following data are from europium-conjugate immunosorbent assay (EuLISA) measuring the 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 13B] Data from EuLISA measuring the binding of 70% pure VH6 / VL3-p5 (6-3-p5), 65% pure VH6 / VL3-p5R (6-3-p5R), c11-1F4, or VH6 / VL3 to rVλ6Wil fibrils are shown. [Figure 13C] Data from EuLISA measuring the binding of VH9 / VL / 4-p5R to rVλ6Wil fibrils, Per125 wtATTR extract, Ken ATTR extract, SHI ALλ liver extract, or TAL ALκ liver extract are shown. [Figure 13D] Data from EuLISA measuring the binding of VH9 / VL / 4-p5 to rVλ6Wil fibrils, Per125 wtATTR extract, Ken ATTR extract, SHI ALλ liver extract, or TAL ALκ liver extract are shown. [Figure 13E] The following data from EuLISA measures the binding of c11-1F4, m11-1F4, or VH9 / VL4 to rVλ6Wil fibrils. [Figure 13F] The image shows data from EuLISA measuring the binding of VH6 / VL3-p5 to Sno ATTR extract (dark gray circle) or Ken ATTR extract (light gray circle), and the binding of c11-1F4 to Sno ATTR extract (black square). [Figure 13G] The image shows data from EuLISA measuring the binding of VH6 / VL3-p5R to Per125 wtATTR (gray circle, see label), Sno ATTR extract (dark gray circle), or Ken ATTR extract (light gray circle), and the binding of c11-1F4 to Sno ATTR extract (black square). The x-axis shows the logarithmically converted molar concentration (-log(M)) of the monoclonal antibody, and the y-axis shows the level of binding (femtomole europium). [Figure 14] This graph 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 bound 125I-mIgG-p5, with the binding percentage for each sample shown above the histogram bars. The x-axis shows the types of amyloid extracts tested, from left to right: rVλ6Wil fibrils (71% bound), SNO hereditary (h) ATTR (12% bound), KEN hATTR (15% bound), Per125 wtATTR (31% bound), Per253 wild-type (wt) ATTR (17% bound), ALκ HIG extract (10% bound), ALκ TAL extract (37% bound), ALλ SHI extract (34% bound), and ALλ TYL extract (21% bound). Error bars represent the standard deviation. [Figure 15A]The x-axis shows the 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. The y-axis shows the level of rVλ6Wil fibril uptake (measured in fluorescence units), and the error bars represent the standard deviation. [Figure 15B] The x-axis shows the phagocytosis of pHrodo-red labeled rVλ6Wil fibrils by macrophages, either with THP-1 alone, or in combination with hIgG control, c11-1F4, mIgp5, VH9 / VL4-p5, or VH9 / VL4-p5R, from left to right. The y-axis shows the level of phagocytosis (fluorescence units), and the error bars represent the standard deviation. [Figure 15C] The x-axis shows phagocytosis of pHrodo-red labeled rVλ6Wil fibrils by macrophages in the presence of hIgG control, 5 μg rituxan (chimeric mAb as a negative control), 5 μg c11-1F4, 5 μg VH6 / VL3, 5 μg VH9 / VL4, VH6 / VL3-p5R, or VH6 / VL3-p5, as shown from left to right. The y-axis shows the level of phagocytosis (pHrodo fluorescence), and the error bars represent the standard deviation. [Figure 16] This graph shows the pharmacokinetic (PK) analysis of 125I-labeled VH9 / VL4 antibody administered intravenously to wild-type ("WT") mice. The x-axis represents time after administration in hours, and the y-axis represents the level of blood radioactivity in counts per minute ("cpm"). The curves are fitted using a double exponential decay equation. [Figure 17A] This graph shows the in vivo distribution of 125I-VH9 / VL4-p5 in WT amyloid-free mice. The x-axis, from left to right, represents sampled organs including muscle, liver, pancreas, spleen, left kidney, right kidney, stomach, upper intestine, lower intestine, heart, lungs, and blood. The y-axis represents the in vivo distribution level as the percentage of injected dose per gram of tissue. [Figure 17B]This graph shows the in vivo distribution of 125I-VH9 / VL4-p5R in WT amyloid-free mice. The x-axis, from left to right, represents sampled organs including muscle, liver, pancreas, spleen, left kidney, right kidney, stomach, upper intestine, lower intestine, heart, lungs, and blood. The y-axis represents the in vivo distribution level as the percentage of injected dose per gram of tissue. [Figure 18] This shows data from a peptide capture ELISA measuring the stability of VH6 / VL3-p5 in mouse plasma. 100 nM VH6 / VL3-p5 was added to either PBS at 4°C (darkest gray), PBS at 37°C (medium-low dark gray circles), EDTA anticoagulant plasma at 37°C (medium-low light gray circles), or heparin anticoagulant plasma at 37°C (lightest gray circle). The x-axis represents time after administration in days, and the y-axis represents the antibody bound to the ligand in absorbance units ("au"). [Modes for carrying out the invention]

[0097] Modified immunoglobulins that bind to amyloid are provided herein. In some embodiments, the modified immunoglobulin is a peptide-Ig fusion.

[0098] I. Definition Unless otherwise specified, technical terms are used according to their conventional usage. Definitions of common terms in molecular biology can be found in Benjamin Lewin, Genes IX, Jones and Bartlet, 2008 (ISBN 0763752223), Kendrew et al. (eds.), The Encyclopedia of Molecular Biology, Blackwell Science Ltd., 1994 (ISBN 0632021829), and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, VCH Publishers, Inc., 1995 (ISBN 9780471185710), as well as other similar references. As used herein, the singular forms "a," "an," and "the" refer to both singular and plural forms unless the context clearly indicates otherwise. The abbreviation "e.g." originates from the Latin "exempli gratia" and is used herein to indicate an unrestricted example. Therefore, the abbreviation "e.g." is synonymous with the term "for example." Where used herein, the term "comprises" means "includes."

[0099] In this specification, a range may be expressed as “approximately” from one particular value to and / or “approximately” another particular value. Where such a range is expressed, another aspect includes from one particular value of the range to and / or the other particular value of the range. It will be further understood that each endpoint of a range is important both in relation to the other endpoints and independently of the other endpoints. Similarly, where a value is expressed as an approximation, it will be understood that the preceding use of “approximately” forms another aspect of the particular value. In certain exemplary embodiments, the term “approximately” is understood to mean within the normal acceptable range in the art, e.g., within two standard deviations of the mean. “Approximately” may be understood to mean 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 clearly indicated by the context, all numerical values ​​provided herein may be modified by the term “approximately.” Furthermore, terms used herein, such as “example,” “exemplary,” or “illustrated,” do not imply preference, but rather indicate that the embodiments discussed later are merely examples of the embodiments presented.

[0100] It should be further understood that all base sizes or amino acid sizes, and all molecular weight or molecular mass values ​​given for nucleic acids or polypeptides are approximate and provided for illustrative purposes only. Methods and materials similar to or equivalent to those described herein may be used in carrying out or testing this disclosure, but preferred methods and materials are described below. In the event of any conflict, including a definition of terms, this specification shall prevail. In addition, the materials, methods, and examples are illustrative and not intended to limit the scope of this specification.

[0101] To facilitate an overview of the various embodiments of this disclosure, the following explanations of specific terms are provided.

[0102] Administration: Introduction of the composition into the subject via a selected route. For example, if the selected route is intravenous, the composition is administered by introducing it into the subject's vein. In some embodiments, a peptide is administered to the subject.

[0103] The terms amyloid, amyloid deposits, amyloid fibrils, and amyloid fibers refer to aggregates of insoluble fibrous proteins that share specific structural characteristics. Protein aggregates are formed, for example, by the aggregation of any one of several different proteins and have a tertiary structure consisting of a regular arrangement of β-sheets stacked perpendicular to the fiber axis. See Sunde et al., J.Mol.Biol. (1997) 273:729-39. Abnormal accumulation of amyloid in organs can lead to amyloidosis. Although their development is diverse, all amyloids share common morphological characteristics in that they stain with certain dyes, such as Congo red, and have a characteristic reddish-green birefringent appearance in polarized light after staining. Amyloids also share common ultrastructural features and common X-ray diffraction and infrared spectra.

[0104] 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 syndrome, hereditary cerebral hemorrhage with Dutch-type amyloidosis, and cerebral β-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.

[0105] Amyloidogenicity refers to the production, or tendency, of amyloid deposits. For example, certain soluble monomer proteins can undergo large-scale conformational changes, leading to their aggregation into highly regular, unbranched fibrils 8-10 nm wide, ultimately resulting in the formation of amyloid aggregates. For instance, in humans, more than 30 different proteins have been found to form amyloid deposits (or amyloid). Not all proteins within a diverse class of proteins, such as immunoglobulin light chains, can form amyloid; that is, some proteins are non-amyloidogenic, meaning they do not tend to form amyloid. However, other proteins in this class can form amyloid deposits and are therefore amyloidogenic. Furthermore, some within the class of light chain proteins may be considered more "amyloidogenic" than others based on their ease of forming amyloid fibrils. Certain light chain proteins are considered non-amyloidogenic or hypoamyloidogenic because they do not readily form amyloid fibrils in patients or in vitro.

[0106] Animals: A category that includes living multicellular vertebrates, such as mammals and birds. The term mammal includes both human and non-human mammals. Similarly, the term "subject" includes both human subjects and veterinary subjects. In some examples, a subject is a subject such as a subject suffering from amyloid disease.

[0107] Clearance: The term "clearance" refers to the measurable degree of clearance. This refers to reduction or removal. For example, amyloid deposit clearance as described herein relates to reducing or removing the deposit to a measurable or identifiable extent. Clearance may result in 100% removal, but it is not required to be 100%. Rather, clearance may result in less than 100% removal (e.g., about 10%, 20%, 30%, 40%, 50%, 60%, or more).

[0108] Conjugate: As used herein, the term “conjugate” refers to the product of the combination or linkage of two or more materials, the resulting product having at least two distinct elements, such as at least two domains. The materials being combined may be the same or different. Such combination may be via 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, a single protein having domains corresponding to each of the individually linked proteins.

[0109] Antibodies refer to single-chain, double-chain, and multi-chain proteins and glycoproteins belonging to the classes of polyclonal, monoclonal, chimeric, and heteroimmunoglobulins (monoclonal antibodies are preferred), and also include synthetic and genetically modified variants of these immunoglobulins. "Antibody fragments" include Fab, Fab', F(ab')2, and Fv fragments, as well as any portion of an antibody having specificity for a desired target epitope(s). "Monoclonal antibodies" are antibodies produced by a single clone of B lymphocytes. Monoclonal antibodies are produced by methods known to those skilled in the art (for example, by creating hybrid antibody-forming cells from a fusion of myeloma cells and immune splenocytes).

[0110] An epitope is a site on an antigen recognized by an antibody, and is determined by the specificity of the antibody's amino acid sequence. Epitopes are also called antigenic determinants. For example, an epitope may be part of a recombinant protein recognized by a specific antibody. Furthermore, epitopes can be structural epitopes or linear epitopes.

[0111] A chimeric antibody refers to an antibody that contains sequences derived from two different antibodies, typically from different species. Most typically, a chimeric antibody contains a human antibody fragment and a mouse antibody fragment (generally a human constant region and a mouse variable region).

[0112] Humanized antibodies refer to antibodies derived from non-human antibodies (typically from mice), as well as human antibodies that retain or substantially retain the antigen-binding properties of the parent antibody but have low immunogenicity in humans.

[0113] Complementarity-determining regions, or CDRs, refer to amino acid sequences that together define the binding affinity and specificity of the native Fv region of the native immunoglobulin binding site. The light and heavy chains of immunoglobulins each have three CDRs, designated L-CDR1, L-CDR2, L-CDR3, and H-CDR1, H-CDR2, and H-CDR3, respectively. By definition, the light chain CDRs are bounded by residues at positions 24 and 34 (L-CDR1), 50 and 56 (L-CDR2), and 89 and 97 (L-CDR3), while the heavy chain CDRs are bounded by residues at positions 31 and 35b (H-CDR1), 50 and 65 (H-CDR2), and 95 and 102 (H-CDR3) (Kabat). et al.,(1991)Sequences of Proteins of Immunological Interest,5th Edition,Department of Health and Human Services,Public Health Service,National Institutes of Health,Bethesda(NIH Publication No.91-324 (Using the numbering rules described in 2).

[0114] The framework region refers to the amino acid sequence inserted between CDRs. These portions of the antibody play a role in holding the CDRs in the correct orientation for antigen binding.

[0115] Specificity-determining residues (SDRs) refer to amino acid residues of immunoglobulins that are directly involved in antigen contact.

[0116] The constant region refers to the portion of the antibody molecule that confers effector function. In this invention, the variant antibody includes a constant region derived from human immunoglobulin. The heavy chain constant region can be selected from one of five isotypes: alpha, delta, epsilon, gamma, or mu. Various subclasses of the heavy chain (e.g., the IgG subclass of the heavy chain) are responsible for different effector functions. Therefore, by selecting the desired heavy chain constant region, a humanized antibody with the desired effector function can be produced. The light chain constant region may be kappa-type or lambda-type (preferably kappa-type).

[0117] Effective dose or therapeutic effective dose: The amount of drug sufficient to prevent, treat (including prevention), reduce, and / or alleviate any symptom and / or underlying cause of any disorder or disease (e.g., to prevent or suppress amyloidosis). In some embodiments, the “effective dose” is sufficient to reduce or eliminate the symptoms of the disease. An effective dose may be administered in more than one dose. For example, an effective dose of peptide is sufficient to bind to amyloid. The peptide may be effective when administered parenterally, for example, in amounts greater than about 1 μg to about 30 mg / kg of body weight.

[0118] Regulatory sequences: Nucleic acid sequences that regulate the expression of a heterologously ligated nucleic acid sequence. Regulatory sequences are ligated to nucleic acid sequences if they control and regulate the transcription and proper translation of the nucleic acid sequence. Thus, regulatory sequences may include appropriate promoters, enhancers, transcriptional terminators, pre-start codons (ATGs) of protein-coding genes, intron splicing signals, maintenance of the correct reading frame of that gene to enable proper translation of mRNA, and stop codons. The term “regulatory sequence” is intended to include at least components whose presence can affect expression, and may also include additional components whose presence is advantageous (e.g., leader sequences and fusion partner sequences). Regulatory sequences may include promoters.

[0119] A promoter is the smallest sequence sufficient to induce transcription. It also includes promoter elements sufficient to make promoter-dependent gene expression cell-type specific, tissue-specific, or induced by external signals or drugs, and such elements may be located in the 5' or 3' region of the gene. Both constitutive and inductive promoters are included (e.g., Bitter et al., Methods). See in Enzymology 153:516-544, 1987). For example, in the case of cloning in bacterial systems, inducible promoters such as bacteriophage λ pL, plac, ptrp, ptac (ptrp-lac hybrid promoter) can be used. In one embodiment, in the case of cloning in mammalian cell systems, promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or promoters derived from mammalian viruses (e.g., retroviral long-chain terminal repeats, adenovirus late promoter, vaccinia virus 7.5K promoter) can be used. Promoters produced by recombinant DNA or synthetic techniques can also be used to provide transcription of nucleic acid sequences. Polynucleotides can be inserted into an expression vector containing a promoter sequence that promotes efficient transcription of the inserted gene sequence in the host. The expression vector typically has an origin of replication, It includes a promoter, as well as specific nucleic acid sequences that enable phenotypic selection of transformed cells.

[0120] Suppression: Reducing to a measurable degree. Suppression does not require, for example, a complete loss or complete cessation of the function of the measured embodiment. For example, suppressing plaque formation may mean stopping further plaque growth, slowing further plaque growth, or reducing the size of the plaque.

[0121] Disease suppression or treatment: Suppressing the complete onset of a disease or condition (e.g., suppressing amyloidosis). "Treatment" refers to a therapeutic intervention that improves signs or symptoms after a disease or pathological condition has begun to develop. The term "alleviate" refers to any observable beneficial effect of treatment with respect to a disease or pathological condition. Beneficial effects may be demonstrated, for example, by delaying the onset of clinical symptoms of the disease in a susceptible subject, reducing the severity of some or all clinical symptoms of the disease, delaying disease progression, improving the subject's overall health or well-being, 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 who shows no signs of the disease or only early signs, with the aim of reducing the risk of developing the condition.

[0122] With regard to amyloid deposition formation, "inhibition" refers to preventing a reduction in amyloid deposition formation, such as compared to a control. For example, inhibition can result in a reduction of approximately 10%, 20%, 30%, 40%, 50%, 60%, or more in amyloid deposition compared to a control.

[0123] A label refers to any detectable compound or composition that is directly or indirectly conjugated to another molecule, thereby facilitating its detection. Specific, non-limiting examples of labels include fluorescent tags, chemiluminescent tags, haptens, enzyme conjugates, and radioisotopes. A protein that is "detectably labeled" means, for example, that the presence of the protein can be determined by a label associated with the protein.

[0124] 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, separately produced, or purified from other biological components of the cell of an organism in which it naturally exists (e.g., other chromosomes and extrachromosomal DNA and RNA, as well as proteins). Thus, “isolated” nucleic acids, peptides, and proteins include nucleic acids and proteins purified by standard purification methods. This term also includes nucleic acids, peptides, and proteins prepared by recombinant expression in cells, as well as chemically synthesized peptides and nucleic acids. The terms “isolated” or “purified” do not require absolute purity and are rather intended as relative terms. Thus, for example, an isolated peptide preparation may contain a higher concentration of the peptide or protein than that present in its natural environment within the cell. Preferably, the preparation is purified such that the protein or peptide represents at least 50% of the total peptide or total 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).

[0125] Linking: As used herein, the terms “join,” “joined,” “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 covalently linked, for example, in a recombinant fusion protein, with or without intervening sequences or domains. Linking can be used to ligate to another protein domain. Linking also includes incorporating two sequences together, for example, by placing two nucleic acid sequences together on the same nucleic acid chain so that the sequences are expressed together.

[0126] Nucleic acids: Polymers consisting of nucleotide units (ribonucleotides, deoxyribonucleotides, related naturally occurring structural variants, and their non-naturally occurring synthetic analogs) linked via phosphodiester bonds, related naturally occurring structural variants, and their non-naturally occurring synthetic analogs. Therefore, this term includes nucleotide polymers in which nucleotides and the links between them contain non-naturally occurring synthetic analogs, such as, but are not limited to, phosphorothioates, phosphoramidates, methylphosphonates, chiral methylphosphonates, 2-O-methylribonucleotides, and peptide nucleic acids (PNAs). Such polynucleotides can be synthesized, for example, using automated DNA synthesizers. The term "oligonucleotide" typically refers to short polynucleotides of generally about 50 nucleotides or less. If a nucleotide sequence is represented by a DNA sequence (i.e., A, T, G, C), it will be understood that RNA sequences (i.e., A, U, G, C) in which "T" is replaced by "U" are also included.

[0127] Nucleotides include, but are not limited to, monomers containing a base linked to a sugar (e.g., pyrimidines, purines, or their synthetic analogs) or a base linked to an amino acid (e.g., peptide nucleic acid (PNA)). A nucleotide is one monomer in a polynucleotide. A nucleotide sequence refers to the sequence of bases in a polynucleotide.

[0128] Conventional notation is used herein to describe nucleotide sequences: the left end of a single-stranded nucleotide sequence is called the 5' end, and the leftward direction of a double-stranded nucleotide sequence is called the 5' direction. The direction of nucleotide addition from 5' to 3' into a nascent RNA transcript is called the transcription direction. The DNA strand having the same sequence as the mRNA is called the "coding strand," and the sequence on the DNA strand having the same sequence as the mRNA transcribed from that DNA and located 5' relative to the 5' end of the RNA transcript is called the "upstream sequence." The sequence on the DNA strand having the same sequence as the RNA and located 3' relative to the 3' end of the coding RNA transcript is called the "downstream sequence."

[0129] cDNA refers to DNA that is complementary to or identical to mRNA, either in single-stranded or double-stranded form.

[0130] To code refers to the inherent properties of a particular sequence of nucleotides in a polynucleotide (e.g., a gene, cDNA, or mRNA), which serve as a template for the synthesis in biological processes of other polymers and macromolecules having either a defined sequence of nucleotides (e.g., rRNA, tRNA, and mRNA) or a defined sequence of amino acids, or the biological properties derived therefrom. Thus, a gene codes for a protein if the transcription and translation of the mRNA produced by that gene produces a protein in a cell or other biological system. Both the coding strand of a gene or cDNA (whose nucleotide sequence is identical to the mRNA sequence and is usually provided in the sequence listing) and the non-coding strand (used as a template for transcription) may be said to code for a protein or other product of that gene or cDNA. Unless otherwise specified, “nucleotide sequences coding an amino acid sequence” include all nucleotide sequences coding the same amino acid sequence, including degenerate versions of each other. Nucleotide sequences coding proteins and RNA may contain introns.

[0131] Recombinant nucleic acids refer to nucleic acids that have nucleotide sequences that are not naturally linked together. This includes nucleic acid vectors (e.g., adenovirus vectors) containing amplified or constructed nucleic acids that can be used to transform suitable host cells. Host cells containing recombinant nucleic acids are called “recombinant host cells.” Genes are then expressed in the recombinant host cells to produce “recombinant polypeptides,” etc. Recombinant nucleic acids can also perform non-coding functions (promoters, origins of replication, ribosome binding sites, etc.). The first sequence is “antisense” with respect to the second sequence if the polynucleotide whose sequence is the first sequence specifically hybridizes with the polynucleotide whose sequence is the second sequence.

[0132] pharmaceutically acceptable carriers: The pharmaceutically acceptable carriers used are conventional ones. Remington's Pharmaceutical Sciences, by EW Martin, Mack Publishing Co., Easton, PA, 19th Edition (1995) describes compositions and formulations suitable for the pharmaceutically acceptable delivery of the fusion proteins disclosed herein.

[0133] Generally, the properties of the carrier will depend on the specific mode of administration used. For example, parenteral formulations typically contain an injectable fluid that is pharmaceutically and physiologically acceptable (e.g., water, saline, equilibrium salt solution, aqueous dextrose, glycerol, etc., as the vehicle). In the case of solid compositions (e.g., in the form of powder, pills, tablets, or capsules), conventional non-toxic solid carriers may include, for example, pharmaceutical-grade mannitol, lactose, starch, or magnesium stearate. In addition to a biologically neutral carrier, the administered pharmaceutical composition may contain small amounts of non-toxic adjuncts (e.g., wetting or emulsifying agents, preservatives, and pH buffers, etc.), such as sodium acetate or sorbitan monolaurate.

[0134] Polypeptide: A polymer in which monomers are amino acid residues linked together via amide bonds. If the amino acids are α-amino acids, either L-optical isomers or D-optical isomers may be used (L-isomers are preferred). As used herein, the terms “polypeptide” or “protein” are intended to encompass any amino acid sequence and include modified sequences such as glycoproteins. Specifically, the term “polypeptide” is intended to encompass naturally occurring proteins as well as those produced by recombinant or synthetic means. In some examples, the peptide is one or more of the peptides disclosed herein.

[0135] Purified: The term "purified" does not require absolute purity; rather, it is intended as a relative term. Therefore, for example, a purified protein preparation is one in which the reference protein is purer than the protein in its natural environment within a cell or production reaction chamber (as appropriate).

[0136] Recombinant nucleic acids are those that have sequences that do not exist in nature, or sequences that are created by the artificial combination of two differently separated segments of a sequence. This artificial combination is often achieved by chemical synthesis, or more commonly, by the artificial manipulation of isolated segments of nucleic acids (e.g., by genetic engineering techniques).

[0137] Sequence Identity: The similarity between two nucleic acid sequences or two amino acid sequences is expressed in terms of sequence similarity and is called sequence identity. Sequence identity is often measured in terms of the percentage of identity (or similarity or homology), with a higher percentage indicating that the two sequences are more similar.

[0138] Methods for arranging sequences for comparison are well known in the relevant art. Various programs and arrangements For column algorithms, see 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. al.Meth.Mol.Bio.24,307-31,1994.Altschul et al. This is described in al. (J.Mol.Biol.215:403-410,1990), where a detailed discussion of sequence sorting methods and homology calculations is presented.

[0139] 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 is available on the internet for use with the sequence analysis programs blastp, blastn, blastx, tblastn, and tblastx.

[0140] Operatively linked: A first nucleic acid sequence is operably linked to a second nucleic acid sequence when the first nucleic acid sequence is positioned to have a functional relationship with the second nucleic acid sequence. For example, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are adjacent to each other in the same reading frame and, if necessary, are intended to link two protein-coding regions.

[0141] Drug: A compound or composition that, when administered appropriately to a target or cell, can induce a desired therapeutic or preventive effect.

[0142] A vector is a nucleic acid molecule that is introduced into a host cell, thereby producing a transformed host cell. A recombinant DNA vector is a vector containing recombinant DNA. A vector may contain nucleic acid sequences that cause it to replicate in a host cell, such as an origin of replication. A vector may also contain genes and other genetic elements of one or more selectable markers known in the art. A viral vector is a recombinant DNA vector having at least several nucleic acid sequences derived from one or more viruses. The term vector includes plasmids, linear nucleic acid molecules, and those described across adenovirus vectors and adenoviruses as a whole.

[0143] The subjects refer to vertebrates. Vertebrates may be mammals (e.g., humans). The subjects may be human patients. The subjects may be patients who have or are suspected of having a disease or condition and may require treatment or diagnosis, or may require monitoring for the progression of the disease or condition. Patients may also be undergoing therapy that needs to be monitored for its effectiveness. In some exemplary embodiments, the subjects include those with amyloidosis (e.g., Alzheimer's disease, Huntington's disease, or prion disease) or peripheral amyloidosis (as seen in patients with light chain (AL) amyloidosis and type 2 diabetes).

[0144] The act of treating or the term treatment refers to the symptoms of a disease or pathological condition after it has begun to manifest. This refers to therapeutic interventions that alleviate symptoms or conditions. The term “alleviate” refers to any observable beneficial effect of treatment with respect to a disease or pathological condition. Beneficial effects may be demonstrated, for example, by delaying the onset of clinical symptoms of the disease in a susceptible subject, reducing the severity of some or all clinical symptoms of the disease, delaying disease progression, improving the subject’s overall health or well-being, 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 who shows no signs of the disease or only early signs, with the aim of reducing the risk of developing the condition.

[0145] Preferably, non-identical residue positions are distinguished by conserved amino acid substitutions. The term "conserved amino acid substitution" refers to the interchangeability of residues having similar side chains. For example, the group of amino acids with aliphatic side chains are glycine, alanine, valine, leucine, and isoleucine; the group of amino acids with aliphatic-hydroxyl side chains are serine and threonine; the group of amino acids with amide-containing side chains are asparagine and glutamine; the group of amino acids with aromatic side chains are phenylalanine, tyrosine, and tryptophan; the group of amino acids with basic side chains are lysine, arginine, and histidine; and the group of amino acids with sulfur-containing side chains are cysteine ​​and methionine. Preferred conserved amino acid substituents are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine.

[0146] As discussed herein, slight variations in the amino acid sequence of antibody or immunoglobulin molecules are intended to be included in this disclosure, provided that such variations maintain at least 75%, more preferably at least 80%, 90%, 95%, and most preferably 99%. Specifically, conservative amino acid substitutions are intended. Conservative substitutions are substitutions that occur within a family of related amino acids in their side chains. Genetically encoded amino acids are generally classified into the following families: (1) acidic amino acids are aspartic acid and glutamic acid; (2) basic amino acids are lysine, arginine, and histidine; (3) nonpolar amino acids are alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; and (4) uncharged polar amino acids are glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Examples of hydrophilic amino acids include arginine, asparagine, aspartic acid, glutamine, glutamic acid, histidine, lysine, serine, and threonine. Examples of hydrophobic amino acids include alanine, cysteine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, tyrosine, and valine. Other families of amino acids include (i) the aliphatic hydroxyl 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, single substitutions of leucine with isoleucine or valine, aspartic acid with glutamic acid, or threonine with serine, or similar substitutions of structurally related amino acids, especially when the substitution does not involve amino acids within a framework site, are reasonably expected to have little effect on the binding or properties of the resulting molecule. Whether amino acid changes result in functional peptides can be easily determined by assaying the specific activity of polypeptide derivatives. The assay is described in detail herein.Fragments or analogues of antibody or immunoglobulin molecules can be readily prepared by those skilled in the art. Preferred amino and carboxyl termini of the fragment or analogue arise near the boundaries of the functional domain. Structural and functional domains can be identified by comparing nucleotide and / or amino acid sequence data with public or proprietary sequence databases. Preferably, sequence motifs arising within other proteins of known structure and / or function. Alternatively, computerized comparison methods are used to identify the expected protein structural domains. Methods for identifying protein sequences that fold into known three-dimensional structures are known (Bowie et al. Science 253:164 (1991)). Thus, the above examples demonstrate that those skilled in the art can recognize sequence motifs and structural structures that can be used to define structural and functional domains according to the present invention.

[0147] Preferred amino acid substitutions are those that (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for protein complex formation, (4) alter binding affinity, or (5) impart or modify other physicochemical or functional properties of such analogues. Analogues may include various mutant proteins of a given sequence other than naturally occurring peptide sequences. For example, one or more amino acid substitutions (preferably conserved amino acid substitutions) may occur in naturally occurring sequences (preferably in the polypeptide portion outside of the domain(s) that form intermolecular contacts). Conservative amino acid substitutions do not substantially alter the structural features of the parent sequence (for example, the substituted amino acids do not tend to disrupt helices that occur in the parent sequence, nor do they tend to disrupt other types of secondary structures that characterize the parent sequence). Examples of recognized secondary and tertiary structures of polypeptides in the art include Proteins, Structures and Molecular Principles (Creighton, Ed., WH Freeman and Company, New York (1984)), Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, NY (1991)), and Thornton This is described in et al. Nature 354:105 (1991).

[0148] Except for CDR1 in VH, CDRs generally contain amino acid residues that form a hypervariable loop. CDRs also contain a "specificity-determining residue" or "SDR," which is the residue that contacts the antigen. SDRs are contained within a region of CDR called abbreviated CDR or a-CDR. Exemplary CDRs (a-CDR-L1, a-CDR-L2, a-CDR-L3, a-CDR-H1, a-CDR-H2, and a-CDR-H3) arise at amino acid residues 31-34 (L1), 50-55 (L2), 89-96 (L3), 31-35B (H1), 50-58 (H2), and 95-102 (H3) (see Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)).

[0149] "Framework" or "FR" refers to residues in the variable domain other than the CDR residue. The variable domain FR generally consists of the following four FR domains: FR1, FR2, FR3, and FR4. Therefore, the CDR sequence and FR sequence generally appear in the following VH (or VL) sequence: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4, or FR1-CDR-H1(L1)-FR2-CDR-H2(L2)-FR3-CDR3-H3(L3)-FR4.

[0150] II. Modified immunoglobulins and antibody-peptide fusion proteins A. Modified immunoglobulins In certain exemplary embodiments, modified immunoglobulins targeting amyloid are provided. Such modified immunoglobulins include, for example, amyloid-reactive peptides, which are ligated to immunoglobulin (Ig) by means of a fragment, via elongation of the N-terminus of an Ig light chain protein in the antigen-binding (Fab) region, or via the C-terminus of the heavy chain, thereby forming a peptide-Ig fusion. Modified immunoglobulins can be used, for example, by administering the modified immunoglobulin to a subject suffering from amyloidosis. It can treat the target. In some embodiments, the modified immunoglobulin is a fusion protein containing an antibody linked to an amyloid-reactive peptide.

[0151] In some embodiments, the modified immunoglobulin comprises an antibody linked to a peptide. In some embodiments, the modified immunoglobulin comprises an antibody containing 1, 2, 3, 4, 5, or 6 CDRs of antibody 11-1F4. In some embodiments, the antibody comprises the VH and / or VL of antibody 11-1F4. In some embodiments, the antibody comprises the heavy and / or light chains of antibody 11-1F4, and the antibody is linked to a peptide.

[0152] In certain embodiments, the modified immunoglobulin comprises an antibody, the antibody comprising VH comprising (a) CDR-H1 containing the amino acid sequence of SEQ ID NO: 17, (b) CDR-H2 containing the amino acid sequence of SEQ ID NO: 18, and (c) CDR-H3 containing the amino acid sequence of SEQ ID NO: 19, and the antibody is linked to a peptide.

[0153] In certain embodiments, the modified immunoglobulin comprises an antibody, the antibody comprising VL comprising (a) CDR-L1 containing the amino acid sequence of SEQ ID NO: 20, (b) CDR-L2 containing the amino acid sequence of SEQ ID NO: 21, and (c) CDR-L3 containing the amino acid sequence of SEQ ID NO: 22, and the antibody is linked to a peptide.

[0154] In one embodiment, the modified immunoglobulin comprises an antibody containing VL, which has the amino acid sequence of SEQ ID NO: 16, and VH, which has the amino acid sequence of SEQ ID NO: 15, and the antibody is linked to a peptide.

[0155] In another embodiment, the modified immunoglobulin comprises an antibody comprising VH, which comprises CDR-H1 containing the amino acid sequence of SEQ ID NO: 17, CDR-H2 containing the amino acid sequence of SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence of SEQ ID NO: 19; and VL, which comprises CDR-L1 containing the amino acid sequence of SEQ ID NO: 20, CDR-L2 containing the amino acid sequence of SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence of SEQ ID NO: 22, and the antibody is linked to a peptide. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing any of the amino acid sequences listed in Table 1. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 2.

[0156] In another embodiment, the modified immunoglobulin comprises an antibody comprising VH CDR1, VH CDR2, and VH CDR3 of VH having the sequence described in SEQ ID NO: 15, and VL CDR1, VL CDR2, and VL of VL having the sequence described in SEQ ID NO: 16, wherein the antibody is linked to a peptide. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing any of the amino acid sequences listed in Table 1. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 2.

[0157] In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide. In some embodiments, the modified immunoglobulin comprises a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 without a C-terminal lysine residue and VL with the amino acid sequence of SEQ ID NO: 16, the antibody being linked to a peptide.

[0158] In another embodiment, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing one of the amino acid sequences listed in Table 1. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin comprises an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the N-terminus of the light chain or the C-terminus of the heavy chain of the antibody. In some embodiments, the antibody also contains a spacer amino acid sequence between the peptide and the N-terminus of the light chain or the C-terminus of the heavy chain. In some embodiments, the peptide is linked to the N-terminus of the light chain. In some embodiments, the peptide is linked to the N-terminus of the heavy chain.

[0159] In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, the heavy chain being linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, the heavy chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, the heavy chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0160] In some embodiments, the modified immunoglobulin comprises an antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the modified immunoglobulin comprises an antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin comprises an antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0161] In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide having one of the amino acid sequences in Table 1.

[0162] In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide having the amino acid sequence of SEQ ID NO: 1. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0163] In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide having the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0164] In some embodiments, the modified immunoglobulins described herein bind to amyloid deposits or amyloid fibrils. In some embodiments, the modified immunoglobulins bind to one or more amyloidogenic peptides of amyloid. In some embodiments, the amyloid bound by the modified immunoglobulins includes amyloidogenic λ6 variable domain protein (Vλ6Wil) or amyloidogenic immunoglobulin light chain (AL), Aβ(1-40) amyloidoid fibrils or amyloidogenic Aβ precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid bound by the modified immunoglobulins includes amyloidogenic immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTR), apolipoprotein This includes protein AI (AApoAI), apolipoprotein AII (AApoAII), gelsolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen A variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP), α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or IAAP, ALκ4, Alλ1, and other amyloidogenic peptides. The amyloidogenic peptides bound by modified immunoglobulins may be proteins, protein fragments, or protein domains. In some embodiments, the amyloid deposit or amyloid fibrils contain recombinant amyloidogenic proteins. In some embodiments, amyloid is part of the pathology of the disease.

[0165] As those skilled in the art will understand, the antigen-binding fragment (or Fab region) is the head of an antibody that naturally interacts with a target antigen. Components of the Fab region allow, for example, an antibody to bind to a specific ligand and, through that interaction, further activate the immune system. For the antibody isotypes IgG, IgA, IgD, IgE, and IgM, Ig consists of two proteins, a heavy chain and a light chain, which interact in pairs to form intact Ig, containing two heavy chains and two light chains. Both the heavy and light chains are further divided into variable and constant domains (lightly variable and heavily variable domains containing the Fab functional region, as well as the heavy chain forming a fragment crystalline (Fc) domain that interacts with cellular receptors and complement). The Fc region of Ig has highly conserved N-glycosylation sites.

[0166] In certain exemplary embodiments, one or more of the peptides shown in Table 1 below may be ligated to an Ig antibody or its functional fragment via the N-terminus of the light chain protein or the C-terminus of the heavy chain, thereby forming a modified immunoglobulin. That is, any of the sequences specified below in Table 1 can be ligated independently or simultaneously to the heavy chain or light chain of an Ig antibody or its functional fragment to form a peptide-Ig conjugate. For example, two of the amyloid-reactive peptides can be ligated to a single Ig antibody by ligating the amino acid sequence of the amyloid-reactive peptide to the N-terminus of an Ig light chain protein. [Table 1]

[0167] While we do not wish to be bound by any particular theory, it is thought that the peptide domain of the peptide-Ig conjugate, when administered to a subject, targets the modified immunoglobulin to amyloid deposits. The Fc domain then triggers an immune response at the amyloid site, thereby leading to the removal of amyloid by means of opsonization. In addition, modified immunoglobulin is thought to have a longer half-life than amyloid-reactive peptide alone. For example, the circulating half-life of IgG in humans is approximately 21 days, while the half-life of amyloid-reactive peptide alone in humans is approximately 11 hours. Therefore, Ig enhances the half-life of circulating modified immunoglobulin. In certain exemplary embodiments, contact of amyloid deposits with modified immunoglobulin results in a half-life of approximately 10%, 20%, and 30% compared to contact of amyloid deposits with amyloid-reactive peptide alone. This results in a half-life that is increased by 40%, 50%, 60%, 70%, 80%, or more. Therefore, when modified immunoglobulins are administered to a target, they can exert an immunostimulatory effect for a longer period at the site of amyloid deposition, thereby increasing the immune response at the site of amyloid deposition.

[0168] In some embodiments, the amyloid-reactive peptide of the modified immunoglobulin peptide described herein comprises an amino acid sequence that is at least 80%, 85%, 90%, or more identical (for example, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the amino acid sequence described as any one of SEQ ID NOs: 1 to 14. In some embodiments, the amyloid-reactive peptide conjugated to an Ig antibody or its functional fragment comprises or may consist of about 10 to about 55 amino acids. The amyloid-reactive peptides of the present invention may contain, 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 No. WO2016032949 (the entire application of which is incorporated herein by reference).

[0169] The amino acids that form all or part of the amyloid-reactive peptide bound to the Ig antibody or a fragment thereof may be naturally occurring amino acids, naturally occurring amino acids, post-translationally modified amino acids, enzymatically synthesized amino acids, derivatized amino acids, stereoisomers and modifications such as constructs or structures designed to mimic amino acids. The amino acids that form the peptide of the present invention may be one or more of the 20 common amino acids found in naturally occurring proteins, or one or more of modified amino acids and atypical amino acids. Modified immunoglobulins may be prepared by any technique known to those skilled in the art (including chemical synthesis or recombinant means using standard molecular biological techniques).

[0170] In certain exemplary embodiments, recombinant DNA technology can be used to clone the nucleotide sequence encoding the peptide of the present invention, fuse it to an Ig light chain, insert it into an expression vector, transform or transfect it into a suitable host cell, and culture it under conditions suitable for expression (see Examples). The peptide-Ig light chain fusion is then isolated. Advantageously, as those skilled in the art will understand in view of this disclosure, any peptide sequence can be ligated to an Ig antibody using the methods described herein. That is, amyloid-reactive peptides are used as an example of peptides ligated to an Ig antibody, but the method of ligating a peptide to an Ig antibody (e.g., to the N-terminus of an Ig light chain protein and / or to the N-terminus and / or C-terminus of an Ig heavy chain protein) can be used for a variety of different peptides to ligate a peptide to an Ig antibody.

[0171] In certain exemplary embodiments, multiple identical or different peptides may be ligated to a single Ig antibody or a functional fragment thereof. For example, a first expression vector may include a light chain nucleic acid sequence that is integrated with a nucleic acid sequence encoding peptide A, with the nucleic acid sequence of peptide A positioned in the vector so that peptide A is ligated to the N-terminus of a light chain protein and expressed. Furthermore, a second expression vector may include a heavy chain nucleic acid sequence that is integrated with a nucleic acid sequence encoding peptide B, with the nucleic acid sequence of peptide B positioned in the vector so that peptide B is ligated to the N-terminus of a light chain protein and expressed.

[0172] In such exemplary embodiments, when both expression vectors are expressed in the same cell, the resulting Ig protein has one peptide A sequence at the N-terminus of each light chain (a total of two peptides). A) can have peptide B at the N-terminus of the heavy chain. In certain exemplary embodiments, the vector may contain peptide C at the C-terminus, thereby yielding an antibody having two peptide A sequences (one on each light chain), a peptide B sequence at the N-terminus of the heavy chain, and a peptide C sequence linked to the C-terminus of the heavy chain. Thus, as those skilled in the art will understand by this disclosure, expression vectors can be modified to modify immunoglobulins to have the same or different combinations of proteins. In a particular example using amyloid-reactive peptides, the modified immunoglobulin may contain two p5 protein sequences (SEQ ID NO: 1), i.e., one at the N-terminus of each light chain. In other exemplary embodiments, the peptide linked to the immunoglobulin may have affinity for a ligand and can therefore be used to detect the ligand.

[0173] In certain exemplary embodiments, modified immunoglobulins can be obtained by isolation or purification. Protein purification techniques, to some extent, involve homogenizing and crudely fractionating cells, tissues, or organs into peptide and non-peptide fractions. Other protein purification techniques include, for example, precipitation with ammonium sulfate, polyethylene glycol (PEG), antibodies, etc., or precipitation by thermal denaturation, followed by centrifugation, chromatography steps (e.g., ion exchange, gel filtration, reversed phase, hydroxyl apatite, and affinity chromatography), isoelectric focusing, gel electrophoresis, for example, polyacrylamide gel electrophoresis, and combinations of these techniques and other techniques.

[0174] Various chromatographic techniques include, but are not limited to, ion exchange chromatography, gel exclusion chromatography, affinity chromatography, immunoaffinity chromatography, and reversed-phase chromatography. Particularly efficient methods for purifying peptides are high-performance liquid chromatography (FPLC) or high-performance liquid chromatography (HPLC). In certain exemplary embodiments, the Fc domain may be linked to the amyloid-reactive peptide via the GGGYS linker sequence (SEQ ID NO: 27).

[0175] In certain embodiments, the modified immunoglobulin may include an amino acid spacer sequence between the N-terminus of the light chain or the C-terminus of the heavy chain and the amyloid-reactive peptide. In certain embodiments, the peptide-Ig conjugate may include an amino acid spacer sequence between the N-terminus of the peptide and the leader sequence required for the secretion of the Ig peptide from the reagent-expressing cell. In some embodiments, the spacer peptide may contain or consist of about 3 to about 55 amino acids. The spacer peptide of the present invention contains or may consist of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 amino acids. As used herein, if nucleic acid sequences or amino acid sequences are in close proximity to each other in sequence, then a nucleic acid sequence or amino acid sequence is "adjacent" to another nucleic acid sequence or amino acid sequence. For example, two nucleic acid sequences may be adjacent to each other as described herein, but may still contain an intervening spacer sequence.

[0176] B. Antibody-peptide fusion protein Also provided herein are antibody-peptide fusion proteins that target amyloid. Such antibody-peptide fusion proteins include, for example, amyloid-reactive peptides, which are ligated to immunoglobulin (Ig) for example, via a fragment, via elongation of the N-terminus of an Ig light chain protein in the antigen-binding (Fab) region, or via the C-terminus of the heavy chain, thereby forming a peptide-Ig fusion. Antibody-peptide fusion proteins can be used to treat subjects suffering from amyloidosis, for example, by administering the antibody-peptide fusion protein to the subject.

[0177] In some embodiments, the antibody-peptide fusion protein includes an antibody linked to a peptide. In some embodiments, the antibody-peptide fusion protein includes an antibody containing 1, 2, 3, 4, 5, or 6 CDRs of the antibody, as shown in Table 3.

[0178] In a particular embodiment, the antibody-peptide fusion protein comprises an antibody, the antibody comprising VH comprising (a) CDR-H1 having the amino acid sequence of SEQ ID NO: 17, (b) CDR-H2 having the amino acid sequence of SEQ ID NO: 18, and (c) CDR-H3 having the amino acid sequence of SEQ ID NO: 19, and the antibody is linked to the peptide.

[0179] In certain embodiments, the antibody-peptide fusion protein comprises an antibody, the antibody comprising a VL comprising (a) CDR-L1 containing the amino acid sequence of SEQ ID NO: 20, (b) CDR-L2 containing the amino acid sequence of SEQ ID NO: 21, and (c) CDR-L3 containing the amino acid sequence of SEQ ID NO: 22, and the antibody is linked to the peptide.

[0180] In one embodiment, the antibody-peptide fusion protein comprises an antibody containing VL, which has the amino acid sequence of SEQ ID NO: 16, and VH, which has the amino acid sequence of SEQ ID NO: 15, wherein the antibody is linked to the peptide.

[0181] In another embodiment, the antibody-peptide fusion protein comprises an antibody comprising VH, which includes CDR-H1 containing the amino acid sequence of SEQ ID NO: 17, CDR-H2 containing the amino acid sequence of SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence of SEQ ID NO: 19; and VL, which includes CDR-L1 containing the amino acid sequence of SEQ ID NO: 20, CDR-L2 containing the amino acid sequence of SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence of SEQ ID NO: 22, and the antibody is linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide containing 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 containing 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 containing the amino acid sequence of SEQ ID NO: 2.

[0182] In another embodiment, the antibody-peptide fusion protein comprises an antibody comprising VH CDR1, VH CDR2, and VH CDR3 of VH having the sequence described in SEQ ID NO: 15, and VL CDR1, VL CDR2, and VL of VL having the sequence described in SEQ ID NO: 16, and the antibody is linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide having one 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 having 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 having the amino acid sequence of SEQ ID NO: 2.

[0183] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 without a C-terminal lysine residue and a VL with the amino acid sequence of SEQ ID NO: 16, the antibody being linked to the peptide.

[0184] In another embodiment, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide. In some embodiments, the antibody-peptide fusion protein is listed in Table 1. The antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide containing one of the amino acid sequences of the specified sequence. In some embodiments, the antibody-peptide fusion protein comprises an antibody linked to an amyloid-reactive peptide containing 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 containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the N-terminus of the light chain or the C-terminus of the heavy chain of the antibody. In some embodiments, the antibody also includes a spacer amino acid sequence between the peptide and the N-terminus of the light chain or the C-terminus of the heavy chain. In some embodiments, the peptide is linked to the N-terminus of the light chain.

[0185] In some embodiments, the antibody-peptide fusion protein comprises an antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0186] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0187] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide containing any of the amino acid sequences in Table 1.

[0188] In some embodiments, the antibody-peptide fusion protein comprises an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0189] In some embodiments, the antibody-peptide fusion protein comprises an antibody and a heavy chain comprising VH containing the amino acid sequence of SEQ ID NO: 15, and a light chain comprising VL containing the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0190] In some embodiments, the antibody-peptide fusion protein described herein binds to amyloid deposits or amyloid fibrils. In some embodiments, the antibody-peptide fusion protein binds to one or more amyloidogenic peptides of amyloid. In some embodiments, the amyloid bound by the antibody-peptide fusion protein includes amyloidogenic λ6 variable domain protein (Vλ6Wil) or amyloidogenic immunoglobulin light chain (AL), Aβ(1-40) amyloidoid fibrils or amyloidogenic Aβ precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid bound by the antibody-peptide fusion protein includes amyloidogenic immunoglobulin heavy chain (AH), β2-microglobulin (Aβ2M), transthyretin variant (ATTR), apolipoprotein AI (AApoAI), apolipoprotein The amyloidogenic peptides include chloroform AII (AApoAII), gelsolin (AGel), lysozyme (ALys), leukocyte chemotactic factor (ALect2), fibrinogen A variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP), α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or IAAP, ALκ4, Alλ1, and other amyloidogenic peptides. The amyloidogenic peptides conjugated by the antibody-peptide fusion protein may be proteins, protein fragments, or protein domains. In some embodiments, the amyloid deposits or amyloid fibrils include recombinant amyloidogenic proteins. In some embodiments, amyloid is part of the pathology of the disease.

[0191] In some embodiments, the antibodies provided herein bind specifically to amyloid light chain fibrils. In some embodiments, the amyloid-reactive peptide binds 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 conjugated by the antibody-peptide fusion protein 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 A variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP), α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or IAAP, ALκ4, Alλ1, or other amyloidogenic peptides. In some embodiments, the amyloid-reactive peptide binds to a heparan sulfate glycosaminoglycan.

[0192] As those skilled in the art will understand, the antigen-binding fragment (or Fab region) is the head of an antibody that naturally interacts with a target antigen. Components of the Fab region allow, for example, an antibody to bind to a specific ligand and, through that interaction, further activate the immune system. For the antibody isotypes IgG, IgA, IgD, IgE, and IgM, Ig consists of two proteins, a heavy chain and a light chain, which interact in pairs to form intact Ig, containing two heavy chains and two light chains. Both the heavy and light chains are further divided into variable and constant domains (lightly variable and heavily variable domains containing the Fab functional region, as well as the heavy chain forming a fragment crystalline (Fc) domain that interacts with cellular receptors and complement). The Fc region of Ig has highly conserved N-glycosylation sites.

[0193] In certain exemplary embodiments, one or more of the peptides shown in Table 1 below may be ligated to an Ig antibody or its functional fragment via the N-terminus of the light chain protein or the C-terminus of the heavy chain, thereby forming an antibody-peptide fusion protein. That is, any of the sequences specified below in Table 1 can be ligated independently or simultaneously to the heavy chain or light chain of an Ig antibody or its functional fragment to form a peptide-Ig conjugate. For example, two of the amyloid-reactive peptides can be ligated to a single Ig antibody by ligating the amino acid sequence of the amyloid-reactive peptide to the N-terminus of an Ig light chain protein.

[0194] While we do not wish to be bound by any particular theory, the peptide domain of a peptide-Ig conjugate, when administered to a target, can affect the antibody-peptide fusion protein. It is thought that the antibody-peptide fusion protein targets amyloid deposits. Subsequently, the Fc domain triggers an immune response at the amyloid site, thereby leading to the removal of amyloid through opsonization or other means. In addition, the antibody-peptide fusion protein is thought to have 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. Therefore, Ig enhances the half-life of the circulating antibody-peptide fusion protein. In certain embodiment examples, contact of amyloid deposits with the antibody-peptide fusion protein results in a half-life that is approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or more than that of contact of amyloid deposits with the amyloid-reactive peptide alone. Therefore, when administered to a target, antibody-peptide fusion proteins can exert an immunostimulatory effect for a longer period at amyloid deposition sites, thereby increasing the immune response at those sites.

[0195] 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 (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the amino acid sequence described as any one of SEQ ID NOs: 1 to 14. In some embodiments, the amyloid-reactive peptide conjugated to the Ig antibody or its functional fragment comprises or may consist of about 10 to about 55 amino acids. The amyloid-reactive peptides of the present invention may contain, 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 No. WO2016032949 (the entire application of which is incorporated herein by reference).

[0196] The amino acids that form all or part of the amyloid-reactive peptide bound to the Ig antibody or a fragment thereof may be naturally occurring amino acids, naturally occurring amino acids, post-translationally modified amino acids, enzymatically synthesized amino acids, derivatized amino acids, stereoisomers and modifications such as constructs or structures designed to mimic amino acids. The amino acids forming the peptide of the present invention may be one or more of the 20 common amino acids found in naturally occurring proteins, or one or more of modified amino acids and atypical amino acids. Antibody-peptide fusion proteins can be prepared by any technique known to those skilled in the art (including chemical synthesis or recombinant means using standard molecular biological techniques).

[0197] In certain exemplary embodiments, recombinant DNA technology can be used to clone the nucleotide sequence encoding the peptide of the present invention, fuse it to an Ig light chain, insert it into an expression vector, transform or transfect it into a suitable host cell, and culture it under conditions suitable for expression (see Examples). The peptide-Ig light chain fusion is then isolated. Advantageously, as those skilled in the art will understand in view of this disclosure, any peptide sequence can be ligated to an Ig antibody using the methods described herein. That is, amyloid-reactive peptides are used as an example of peptides ligated to an Ig antibody, but the method of ligating a peptide to an Ig antibody (e.g., to the N-terminus of an Ig light chain protein and / or to the N-terminus and / or C-terminus of an Ig heavy chain protein) can be used for a variety of different peptides to ligate a peptide to an Ig antibody.

[0198] In certain exemplary embodiments, multiple identical or different peptides may be ligated to a single Ig antibody or a functional fragment thereof. For example, the first expression vector may ligate peptide A. The second expression vector may include a light chain nucleic acid sequence that is integrated with the nucleic acid sequence encoding peptide A, and the nucleic acid sequence of peptide A is positioned in the vector so that peptide A is ligated to the N-terminus of the light chain protein and expressed. Furthermore, the second expression vector may include a heavy chain nucleic acid sequence that is integrated with the nucleic acid sequence encoding peptide B, and the nucleic acid sequence of peptide B is positioned in the vector so that peptide B is ligated to the N-terminus of the light chain protein and expressed.

[0199] In such exemplary embodiments, when both expression vectors are expressed in the same cell, the resulting Ig protein may have one peptide A sequence at the N-terminus of each light chain (two peptide A sequences in total) and one peptide B at the N-terminus of the heavy chain. In certain exemplary embodiments, the vector may contain peptide C at its C-terminus, thereby yielding an antibody having two peptide A sequences (one on each light chain), a peptide B sequence at the N-terminus of the heavy chain, and a peptide C sequence linked to the C-terminus of the heavy chain. Thus, as those skilled in the art will understand by this disclosure, expression vectors may be modified to modify immunoglobulins to have the same or different combinations of proteins. In a specific example using amyloid-reactive peptides, an antibody-peptide fusion protein may contain two p5 protein sequences (SEQ ID NO: 1), i.e., one at the N-terminus of each light chain. In other exemplary embodiments, the peptide linked to the immunoglobulin may have affinity for the ligand and can therefore be used to detect the ligand.

[0200] In certain exemplary embodiments, antibody-peptide fusion proteins can be obtained by isolation or purification. Protein purification techniques, to some extent, involve homogenizing and crudely fractionating cells, tissues, or organs into peptide and non-peptide fractions. Other protein purification techniques include, for example, precipitation with ammonium sulfate, polyethylene glycol (PEG), antibodies, etc., or precipitation by thermal denaturation, followed by centrifugation, chromatography steps (e.g., ion exchange, gel filtration, reversed phase, hydroxyl apatite, and affinity chromatography), isoelectric focusing, gel electrophoresis, for example, polyacrylamide gel electrophoresis, and combinations of these techniques and other techniques.

[0201] Various chromatographic techniques include, but are not limited to, ion exchange chromatography, gel exclusion chromatography, affinity chromatography, immunoaffinity chromatography, and reversed-phase chromatography. Particularly efficient methods for purifying peptides are high-performance liquid chromatography (FPLC) or high-performance liquid chromatography (HPLC). In certain exemplary embodiments, the Fc domain may be linked to the amyloid-reactive peptide via the GGGYS linker sequence (SEQ ID NO: 27).

[0202] In certain embodiments, the antibody-peptide fusion protein may include an amino acid spacer sequence between the N-terminus of the light chain or the C-terminus of the heavy chain and the amyloid-reactive peptide. In certain embodiments, the peptide-Ig conjugate may include an amino acid spacer sequence between the N-terminus of the peptide and a leader sequence required for the secretion of the Ig peptide from the reagent-expressing cell. In some embodiments, the spacer peptide may contain or consist of about 3 to about 55 amino acids. The spacer peptide of the present invention contains or may consist of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 amino acids. As used herein, if nucleic acid sequences or amino acid sequences are in close proximity to each other in sequence, then a nucleic acid sequence or amino acid sequence is "adjacent" to another nucleic acid sequence or amino acid sequence. For example, two nucleic acid sequences may be adjacent to each other as described herein, but may still contain an intervening spacer sequence.

[0203] III. Humanized antibodies that bind to human amyloid fibrils Humanized antibodies that bind to human amyloid fibrils are provided herein. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH) containing one or more CDRs of a mouse antibody. In some embodiments, the VH and / or VL are derived from human VH and / or VL sequences (e.g., "human acceptor sequences"). In some embodiments, the VH and / or VL include, for example, amino acid substitutions in the framework region of the VH and / or VL.

[0204] A. Humanized antibodies In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises 1, 2, 3, 4, 5, or 6 CDRs of the antibody, as shown in Table 3. In some embodiments, the humanized antibody comprises CDR-H1, CDR-H2, and CDR-H3, each containing the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequence described in SEQ ID NO: 15, and CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequence described in SEQ ID NO: 16. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20 having one or more conserved amino acid substitutions, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21 having one or more conserved amino acid substitutions, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22 having one or more conserved amino acid substitutions; and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17 having one or more conserved amino acid substitutions, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18 having one or more conserved amino acid substitutions, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19 having one or more conserved amino acid substitutions. In some embodiments, the humanized antibody comprises 1, 2, 3, 4, 5, or 6 CDRs of the antibody shown in Table 3, each having one or more conserved amino acid substitutions.In some embodiments, the humanized antibody comprises CDR-H1, CDR-H2, and CDR-H3, each containing the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequence described in SEQ ID NO: 15, which has one or more conserved amino acid substitutions, and CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequence described in SEQ ID NO: 16, which has one or more conserved amino acid substitutions.

[0205] In some embodiments, a humanized antibody that binds to human amyloid fibrils includes a light chain variable region (VL) and a heavy chain variable region (VH), where VL includes a framework region (FR) and VH includes a framework region (FR). In some embodiments, the framework region is FR1, FR2, FR3, or FR4. In some embodiments, VL includes FR1, FR2, FR3, and FR4 from the N-terminus to the C-terminus. In some embodiments, VH includes FR1, FR2, FR3, or FR4. In some embodiments, VH includes FR1, CDR-H1, FR2, CDR-H2, FR3, CDR-H3, and FR4 from the N-terminus to the C-terminus. In some embodiments, the humanized antibody includes a VL and / or VH containing amino acid substitutions at one or more positions in the framework region (e.g., FR1, FR2, FR3, or FR4) compared to a human acceptor sequence having a transplanted CDR (e.g., SEQ ID NO: 32 or SEQ ID NO: 43).

[0206] In some embodiments, the humanized antibody includes a VL containing an amino acid substitution in FR2, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the amino acid substitution in FR2 is selected from the group consisting of an amino acid substitution at position 36, an amino acid substitution at position 37, and an amino acid substitution at position 46, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in FR2 is selected from the group consisting of Tyr at position 36, Leu at position 37, and Leu at position 46, and the amino acid positions are numbered according to the Kabat numbering system.

[0207] In some embodiments, the humanized antibody includes a VL containing an amino acid substitution in FR3, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the amino acid substitution in FR3 is selected from the group consisting of an amino acid substitution at position 85 and an amino acid substitution at position 87, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in FR3 is selected from the group consisting of Leu at position 85 and Phe at position 87, and the amino acid positions are numbered according to the Kabat numbering system.

[0208] In some embodiments, the humanized antibody includes a VH having an amino acid substitution in FR2, compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the amino acid substitution in FR2 is selected from the group consisting of an amino acid substitution at position 37 and an amino acid substitution at position 48, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in FR2 is selected from the group consisting of Val at position 37 and Leu at position 48, and the amino acid positions are numbered according to the Kabat numbering system.

[0209] In some embodiments, the humanized antibody comprises a VH that includes an amino acid substitution in FR3 as compared to the VH comprising the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the amino acid substitution in FR3 is selected from the group consisting of an amino acid substitution at position 67, an amino acid substitution at position 48, an amino acid substitution at position 71, an amino acid substitution at position 76, an amino acid substitution at position 78, an amino acid substitution at position 79, an amino acid substitution at position 80, an amino acid substitution at position 89, an amino acid substitution at position 93, and an amino acid substitution at position 94, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in FR3 is selected from the group consisting of Leu at position 67, Ser at position 48, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Val at position 93, and Thr at position 94, and the amino acid positions are numbered according to the Kabat numbering system.

[0210] In some embodiments, the humanized antibody includes a VL containing one or more amino acid substitutions at one or more positions of the VL compared to a VL containing the amino acid sequence described in SEQ ID NO: 32, where the amino acid positions are sequentially numbered starting from the N-terminus of SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL containing an amino acid substitution at position 33 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL containing an amino acid substitution at position 34 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL containing an amino acid substitution at position 41 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL containing an amino acid substitution at position 42 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL containing an amino acid substitution at position 51 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL containing an amino acid substitution at position 90 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody includes a VL having an amino acid substitution at position 92, compared to a VL having the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody comprises a VL containing 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the humanized antibody comprises a VL containing 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to VL1, as shown in Table 6A.

[0211] In some embodiments, the humanized antibody comprises a VL that includes one or more amino acid residues at one or more positions of the VL, and the amino acid positions are numbered starting from the N-terminus of the VL according to the numbering of SEQ ID NO: 32. In some embodiments, the humanized antibody comprises a VL that includes Ser, Gln, Glu, His, or Ala at position 33. In some embodiments, the humanized antibody comprises a VL that includes Ala or Val at position 34. In some embodiments, the humanized antibody comprises a VL that includes Tyr at position 41. In some embodiments, the humanized antibody comprises a VL that includes Leu at position 42. In some embodiments, the humanized antibody comprises a VL that includes Leu at position 51. In some embodiments, the humanized antibody comprises a VL that includes Leu at position 90. In some embodiments, the humanized antibody comprises a VL that includes Phe at position 92.

[0212] In some embodiments, the humanized antibody comprises a VH that includes one or more amino acid residues at one or more positions of the VH, and the amino acid positions are numbered starting from the N-terminus of the VH. In some embodiments, the humanized antibody comprises a VH that includes Val at position 37. In some embodiments, the humanized antibody comprises a VH that includes Leu at position 48. In some embodiments, the humanized antibody comprises a VH that includes Leu at position 67. In some embodiments, the humanized antibody comprises a VH that includes Ser at position 68. In some embodiments, the humanized antibody comprises a VH that includes Lys at position 71. In some embodiments, the humanized antibody comprises a VH that includes Ser at position 76. In some embodiments, the humanized antibody comprises a VH that includes Val at position 78. In some embodiments, the humanized antibody comprises a VH that includes Leu at position 79. In some embodiments, the humanized antibody comprises a VH that includes Phe at position 80. In some embodiments, the humanized antibody comprises a VH that includes Thr at position 92. In some embodiments, the humanized antibody comprises a VH that includes Val at position 96. In some embodiments, the humanized antibody comprises a VH that includes Thr at position 97.

[0213] In some embodiments, the humanized antibody comprises a VL containing one or more amino acid residues at one or more positions of the VL, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the humanized antibody comprises a VL containing Tyr at position 36. In some embodiments, the humanized antibody comprises a VL containing Leu at position 37. In some embodiments, the humanized antibody comprises a VL containing Leu at position 46. In some embodiments, the humanized antibody comprises a VL containing Leu at position 85. In some embodiments, the humanized antibody comprises a VL containing Phe at position 87.

[0214] In some embodiments, the humanized antibody comprises a VL containing one or more amino acid residues at one or more positions of the VL, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the humanized antibody comprises a VL containing Tyr at position 36 and Leu at position 37. In some embodiments, the humanized antibody comprises a VL containing Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87. In some embodiments, the humanized antibody comprises a VL containing Leu at position 46 and Phe at position 87.

[0215] In some embodiments, the humanized antibody comprises VH containing one or more amino acid residues at one or more positions of VH, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the humanized antibody comprises VH containing Val at position 37. In some embodiments, the humanized antibody comprises VH containing Leu at position 48. In some embodiments, the humanized antibody comprises VH containing Leu at position 67. In some embodiments, the humanized antibody comprises, The humanized antibody contains VH containing Ser at position 68. In some embodiments, the humanized antibody contains VH containing Lys at position 71. In some embodiments, the humanized antibody contains VH containing Ser at position 76. In some embodiments, the humanized antibody contains VH containing Val at position 78. In some embodiments, the humanized antibody contains VH containing Leu at position 79. In some embodiments, the humanized antibody contains VH containing Phe at position 80. In some embodiments, the humanized antibody contains VH containing Thr at position 89. In some embodiments, the humanized antibody contains VH containing Val at position 93. In some embodiments, the humanized antibody contains VH containing Thr at position 94.

[0216] In some embodiments, the humanized antibody comprises a VH containing one or more amino acid residues at one or more positions of the VH, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the humanized antibody comprises a VH containing Val at position 37 and Leu at position 48. In some embodiments, the humanized antibody comprises a VH containing Leu at position 67, Ser at position 68, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises a VH containing Val at position 37, Leu at position 48, Leu at position 67, and Ser at position 68. In some embodiments, the humanized antibody comprises a VH containing Val at position 37, Leu at position 48, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VH containing Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VH containing Lys at position 71, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VH containing Lys at position 71, Ser at position 76, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VH containing Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the humanized antibody contains VH with Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94.

[0217] In some embodiments, the humanized antibody comprises VL containing Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH containing Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VL containing Leu at position 46 and Phe at position 87, and VH containing Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the humanized antibody comprises VL containing Leu at position 46 and Phe at position 87, and VH containing Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VL containing Leu at position 46 and Phe at position 87, and VH containing Lys at position 71, Ser at position 76, Val at position 93, and Thr at position 94. In some embodiments, the humanized antibody comprises VL containing Leu at position 46 and Phe at position 87, and VH containing Lys at position 71, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94.

[0218] In some embodiments, the humanized antibody contains the amino acid sequence of VL shown in Table 6A. In some embodiments, the humanized antibody contains 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 6A. In some embodiments, VL contains the amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42.

[0219] In some embodiments, the humanized antibody contains the amino acid sequence of VH shown in Table 6B. In some embodiments, the humanized antibody contains 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 shown in Table 6B. In some embodiments, VH contains the amino acid sequence described in the group consisting of SEQ ID NOs: 44 to 63.

[0220] In some embodiments, the humanized antibody comprises VL of VL4 shown in Table 6A and VH of VH9 shown in Table 6B. In some embodiments, VL comprises the amino acid sequence described in SEQ ID NO: 35, and VH comprises the amino acid sequence described in SEQ ID NO: 51.

[0221] In some embodiments, the humanized antibody comprises VL of VL3 shown in Table 6A and VH of VH6 shown in Table 6B. In some embodiments, VL comprises the amino acid sequence described in SEQ ID NO: 34, and VH comprises the amino acid sequence described in SEQ ID NO: 48.

[0222] In some embodiments, the humanized antibody includes the VL of VL4 shown in Table 6A and the VH of VH10 shown in Table 6B. In some embodiments, the VL includes the amino acid sequence described in SEQ ID NO: 35, and the VH includes the amino acid sequence described in SEQ ID NO: 52. In some embodiments, the humanized antibody includes the VL of VL4 shown in Table 6A and the VH of VH8 shown in Table 6B. In some embodiments, the VL includes the amino acid sequence described in SEQ ID NO: 35, and the VH includes the amino acid sequence described in SEQ ID NO: 50. In some embodiments, the humanized antibody includes the VL of VL4 shown in Table 6A and the VH of VH7 shown in Table 6B. In some embodiments, the VL includes the amino acid sequence described in SEQ ID NO: 35, and the VH includes the amino acid sequence described in SEQ ID NO: 69.

[0223] In some embodiments, the humanized antibody is a full-length antibody, a Fab fragment, or an scFv.

[0224] Humanized antibodies with CDR substitutions In some embodiments, the humanized antibody contains one or more CDR substitutions. In some embodiments, the CDR substitution is located within CDR-L1. In some embodiments, the CDR substitution is located within CDR-H2.

[0225] In some embodiments, the humanized antibody comprises VL, which includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22; and VH, which includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. In some embodiments, the humanized antibody comprises VL, which includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 20, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22; and VH, which includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequences described in SEQ ID NOs. 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19. In some embodiments, the humanized antibody comprises a VL containing one or more amino acid residues selected from the group consisting of Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the humanized antibody comprises a VH containing one or more amino acid residues selected from the group consisting of: Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, and L at position 79. eu, Phe at position 80, Thr at position 89, Val at position 93, Thr at position 94 (amino acid positions are numbered according to the Kabat numbering system).

[0226] In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 64, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 65, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 66, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 67, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 68, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 69, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 70, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.

[0227] In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 71, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises the amino acid sequence described in SEQ ID NO: 20 The humanized antibody comprises CDR-L1 containing an amino acid sequence, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 72, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), where VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 73, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 74, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 76, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), where the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 78, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 80, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, and the amino acid sequence described in SEQ ID NO: 21. VH includes CDR-L2 containing the sequence and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 81, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19.

[0228] In some embodiments, the humanized antibody comprises one, two, three, four, five, or six CDRs of the antibodies shown in Table 6C and / or Table 6D.

[0229] In some embodiments, the humanized antibody includes CDR-H1, CDR-H2, and CDR-H3, each containing the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequences described in SEQ ID NOs. 53 to 63. In some embodiments, the humanized antibody includes CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequences described in SEQ ID NOs. 36 to 42.

[0230] In some embodiments, the humanized antibody comprises a VL that contains an amino acid substitution in CDR-L1 as compared to the VL comprising the amino acid sequence set forth in SEQ ID NO: 35. In some embodiments, the amino acid substitution in CDR-L1 is selected from the group consisting of an amino acid substitution at position 28 and an amino acid substitution at position 29, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-L1 is selected from the group consisting of Ser, Gln, Glu, His, or Ala at position 28, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-L1 is selected from the group consisting of Ala or Val at position 29, and the amino acid positions are numbered according to the Kabat numbering system.

[0231] In some embodiments, the humanized antibody comprises a VH that contains an amino acid substitution in CDR-H2 as compared to the VH comprising the amino acid sequence set forth in SEQ ID NO: 51. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of an amino acid substitution at position 54, an amino acid substitution at position 55, or an amino acid substitution at position 64, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of Ser, Gln, Glu, Ala, or His at position 54, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of Ala or Val at position 55, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of Val, Ile, Leu, or Ala at position 64, and the amino acid positions are numbered according to the Kabat numbering system.

[0232] B. Antibody-Peptide Fusion Proteins Comprising Humanized Antibodies Antibody-peptide fusion proteins comprising humanized antibodies that bind to human amyloid fibrils fused to amyloid-reactive peptides are also provided herein. In some embodiments, the antibody-peptide fusion protein comprises humanized antibodies described herein. In some embodiments, the humanized antibody is any one of the humanized antibodies described herein.

[0233] In certain embodiments, the antibody-peptide fusion protein comprises a humanized antibody having a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises CDR-L1 having the amino acid sequence described in SEQ ID NO: 20, CDR-L2 having the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 having the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 having the amino acid sequence described in SEQ ID NO: 17, CDR-H2 having the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 having the amino acid sequence described in SEQ ID NO: 19. Morphologically, the humanized antibody contains 1, 2, 3, 4, 5, or 6 CDRs of the antibody, as shown in Table 3. In some embodiments, the humanized antibody contains CDR-H1, CDR-H2, and CDR-H3, each containing the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequence described in SEQ ID NO: 15, and CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequence described in SEQ ID NO: 16. In some embodiments, the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20 having one or more conserved amino acid substitutions, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21 having one or more conserved amino acid substitutions, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22 having one or more conserved amino acid substitutions; and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17 having one or more conserved amino acid substitutions, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18 having one or more conserved amino acid substitutions, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19 having one or more conserved amino acid substitutions. In some embodiments, the humanized antibody comprises 1, 2, 3, 4, 5, or 6 CDRs of the antibody shown in Table 3, each having one or more conserved amino acid substitutions. In some embodiments, the humanized antibody comprises CDR-H1, CDR-H2, and CDR-H3, each containing the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequence described in SEQ ID NO: 15, which has one or more conserved amino acid substitutions, and CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequence described in SEQ ID NO: 16, which has one or more conserved amino acid substitutions.

[0234] In some embodiments, the humanized antibody comprises VL, which includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22; and VH, which includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, which comprises VL, which includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 20, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22; and VH, which includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19.

[0235] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing an amino acid substitution at FR2 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the amino acid substitution at FR2 is selected from the group consisting of an amino acid substitution at position 36, an amino acid substitution at position 37, and an amino acid substitution at position 46, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution at FR2 is selected from the group consisting of Tyr at position 36, Leu at position 37, and Leu at position 46, and the amino acid positions are numbered according to the Kabat numbering system.

[0236] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing an amino acid substitution at FR3 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the amino acid substitution at FR3 is selected from the group consisting of an amino acid substitution at position 85 and an amino acid substitution at position 87, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution at FR3 is selected from the group consisting of Leu at position 85 and Phe at position 87, and the amino acid positions are numbered according to the Kabat numbering system.

[0237] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing an amino acid substitution at FR2 compared to a VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the amino acid substitution at FR2 is selected from the group consisting of an amino acid substitution at position 37 and an amino acid substitution at position 48, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution at FR2 is selected from the group consisting of Val at position 37 and Leu at position 48, and the amino acid positions are numbered according to the Kabat numbering system.

[0238] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH having an amino acid substitution at FR3 compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the amino acid substitution at FR3 is selected from the group consisting of an amino acid substitution at position 67, an amino acid substitution at position 48, an amino acid substitution at position 71, an amino acid substitution at position 71, an amino acid substitution at position 76, an amino acid substitution at position 78, an amino acid substitution at position 79, an amino acid substitution at position 80, an amino acid substitution at position 89, an amino acid substitution at position 93, and an amino acid substitution at position 94, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in FR3 is selected from the group consisting of Leu at position 67, Ser at position 48, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, Thr at position 89, Val at position 93, and Thr at position 94, and the amino acid positions are numbered according to the Kabat numbering system.

[0239] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing one or more amino acid substitutions at one or more positions of the VL compared to a VL containing the amino acid sequence described in SEQ ID NO: 32, where the amino acid positions are numbered starting from the N-terminus of SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing an amino acid substitution at position 33 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing an amino acid substitution at position 34 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing an amino acid substitution at position 41 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing an amino acid substitution at position 42 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing a VL with an amino acid substitution at position 51 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing a VL with an amino acid substitution at position 90 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing a VL with an amino acid substitution at position 92 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing a VL with 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing a VL with 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to VL1, as shown in Table 6A.

[0240] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH having one or more amino acid substitutions at one or more positions of the VH compared to a VH having the amino acid sequence described in SEQ ID NO: 43, the amino acid position being the N-terminus of SEQ ID NO: 43 The numbers are assigned starting from there. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH having an amino acid substitution at position 37 compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH having an amino acid substitution at position 48 compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH having an amino acid substitution at position 67 compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH having an amino acid substitution at position 68 compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH having an amino acid substitution at position 71 compared to a VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing an amino acid substitution at position 76 compared to VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing an amino acid substitution at position 78 compared to VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing an amino acid substitution at position 79 compared to VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing an amino acid substitution at position 80 compared to VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing an amino acid substitution at position 92 compared to VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH having an amino acid substitution at position 96 compared to a VH having the amino acid sequence described in SEQ ID NO: 43.In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH having an amino acid substitution at position 97 compared to VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions compared to VH having the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions compared to VH1, as shown in Table 6B.

[0241] In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing one or more amino acid residues at one or more positions of the VL, the amino acid positions being numbered starting from the N-terminus of the VL. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Ser, Gln, Glu, His, or Ala at position 33. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Ala or Val at position 34. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Tyr at position 41. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 42. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 51. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 90. In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising a VL containing Phe at position 92.

[0242] In some embodiments, the antibody-peptide fusion comprises a humanized antibody, the humanized antibody comprising VH having one or more amino acid residues at one or more positions of VH, the amino acid positions being VH Numbering begins from the N-terminus. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Val at position 37. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Leu at position 48. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Leu at position 67. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Ser at position 68. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Lys at position 71. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Ser at position 76. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Val at position 78. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Leu at position 79. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Phe at position 80. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Thr at position 92. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Val at position 96. In some embodiments, the antibody-peptide fusion includes a humanized antibody, the humanized antibody containing VH with Thr at position 97.

[0243] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing one or more amino acid substitutions at one or more positions of the VL, the amino acid positions being numbered according to the Kabat numbering system. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Tyr at position 36. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 37. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 46. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 85. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Phe at position 87.

[0244] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing one or more amino acid residues at one or more positions of the VL, the amino acid positions being numbered according to the Kabat numbering system. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Tyr at position 36 and Leu at position 37. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VL containing Leu at position 46 and Phe at position 87.

[0245] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing one or more amino acid residues at one or more positions of the VH, the amino acid positions being numbered according to the Kabat numbering system. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Val at position 37. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Leu at position 48. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Leu at position 67. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Ser at position 68. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Lys at position 71. In some embodiments, the antibody-peptide fusion includes a humanized antibody, and the humanized antibody includes a VH containing Ser at position 76. In some embodiments, the antibody-peptide fusion includes a humanized antibody, and the humanized antibody includes a VH containing Val at position 78. In some embodiments, the antibody-peptide fusion includes a humanized antibody, and the humanized antibody includes a VH containing Leu at position 79. In some embodiments, the antibody-peptide fusion includes a humanized antibody, and the humanized antibody includes a VH containing Phe at position 80. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH containing Thr at position 89. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH containing Val at position 93. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, and the humanized antibody includes a VH containing Thr at position 94.

[0246] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing one or more amino acid residues at one or more positions of the VH, the amino acid positions being numbered according to the Kabat numbering system. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Val at position 37 and Leu at position 48. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Leu at position 67, Ser at position 68, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Val at position 37, Leu at position 48, Leu at position 67, and Ser at position 68. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH containing Val at position 37, Leu at position 48, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH containing Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH containing Lys at position 71, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VH containing Lys at position 71, Ser at position 76, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a VH containing Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94.

[0247] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VL containing Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH containing Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising at position 46 The antibody-peptide fusion protein comprises VL containing Leu and Phe at position 87, and VH containing Lys at position 71, Ser at position 76, Val at position 93, and Thr at position 94. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Lys at position 71, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94.

[0248] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing the amino acid sequence of VL shown in Table 6A. 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 6A. In some embodiments, VL contains the amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42.

[0249] In some embodiments, the antibody-peptide fusion protein includes a humanized antibody containing the amino acid sequence of VH shown in Table 6B. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody, the humanized antibody containing 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 shown in Table 6B. In some embodiments, VH contains the amino acid sequence described in the group consisting of SEQ ID NOs: 44 to 63.

[0250] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing VL of VL4 shown in Table 6A and VH of VH9 shown in Table 6B. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing VL containing the amino acid sequence described in SEQ ID NO: 35 and VH containing the amino acid sequence described in SEQ ID NO: 51.

[0251] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing VL of VL3 shown in Table 6A and VH of VH6 shown in Table 6B. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing VL containing the amino acid sequence described in SEQ ID NO: 34 and VH containing the amino acid sequence described in SEQ ID NO: 48.

[0252] In some embodiments, the antibody-peptide fusion protein includes a humanized antibody comprising the VL of VL4 shown in Table 6A and the VH of VH10 shown in Table 6B. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody comprising the VL comprising the amino acid sequence described in SEQ ID NO: 35 and the VH comprising the amino acid sequence described in SEQ ID NO: 52. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody comprising the VL of VL4 shown in Table 6A and the VH of VH8 shown in Table 6B. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody comprising the VL comprising the amino acid sequence described in SEQ ID NO: 35 and the VH comprising the amino acid sequence described in SEQ ID NO: 50. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody comprising the VL of VL4 shown in Table 6A and the VH of VH7 shown in Table 6B. In some embodiments, the antibody-peptide fusion protein includes a humanized antibody comprising the VL comprising the amino acid sequence described in SEQ ID NO: 35 and the VH comprising the amino acid sequence described in SEQ ID NO: 69.

[0253] In some embodiments, the antibody-peptide fusion protein contains an amyloid-reactive peptide. In some embodiments, the amyloid-reactive peptide contains one or more peptides shown in Table 1. In certain embodiments, the amyloid-reactive peptide contains an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 14. In some embodiments, the amyloid The amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 1. In some embodiments, the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 2. In some embodiments, the amyloid-reactive peptide is positively charged.

[0254] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a light chain. In some embodiments, an amyloid-reactive peptide is fused to the N-terminus of the light chain. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody, the humanized antibody comprising a light chain, and the amyloid-reactive peptide is fused to the N-terminus of the light chain by a linker. In some embodiments, the linker is a peptide linker. In some embodiments, the linker comprises the amino acid sequence GGGYS. In some embodiments, the linker comprises the amino acid sequence described in SEQ ID NO: 27. In some embodiments, the linker is uncharged.

[0255] In some embodiments, one or more of the peptides shown in Table 1 below may be bound to a humanized antibody or its functional fragment via the N-terminus of the light chain protein or the C-terminus of the heavy chain, thereby forming an antibody-peptide fusion protein containing the humanized antibody. That is, any of the following sequences identified in Table 1 can be independently or simultaneously ligated to the heavy or light chain of a humanized antibody or its functional fragment to form an antibody-peptide fusion protein. For example, two amyloid-reactive peptides can be ligated to a single Ig antibody by ligating the amino acid sequence of an amyloid-reactive peptide to the N-terminus of the light chain of a humanized antibody.

[0256] In some embodiments, the antibody-peptide fusion protein includes a light chain, comprising an amyloid-reactive peptide and the light chain from the N-terminus to the C-terminus. In some embodiments, the light chain comprises VL and CL1 from the N-terminus to the C-terminus. In some embodiments, VL is one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein includes a heavy chain, comprising VH, CH1, CH2, and CH3 from the N-terminus to the C-terminus. In some embodiments, VH is one of the VHs described herein.

[0257] In some embodiments, the antibody-peptide fusion protein includes a light chain, comprising an amyloid-reactive peptide, a spacer peptide, and another light chain from the N-terminus to the C-terminus. 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 light chain comprises VL and CL1 from the N-terminus to the C-terminus. In some embodiments, VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein includes a heavy chain, comprising VH, CH1, CH2, and CH3 from the N-terminus to the C-terminus.

[0258] In some embodiments, the antibody-peptide fusion protein comprises, from N-terminus to C-terminus, a secreted 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 light chain comprises, from N-terminus to C-terminus, VL and CL1. In some embodiments, VL is any one of the VLs described herein. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain comprising, from N-terminus to C-terminus, VH, CH1, CH2, and CH3. In some embodiments, VH is any one of the VHs described herein. In some embodiments, VH is any one of the VHs described herein. In some embodiments, the antibody-peptide fusion protein comprises the structure illustrated in Figure 11A or Figure 11B.

[0259] In some embodiments, the antibody-peptide fusion protein includes an amino acid spacer sequence between the N-terminus of the light chain and the amyloid-reactive peptide. In some embodiments, the antibody-peptide fusion protein includes an amino acid spacer sequence between the N-terminus of the peptide and a leader sequence required for the secretion of the antibody-peptide fusion protein from cells expressing the antibody-peptide fusion protein. In some embodiments, the spacer peptide is a flexible spacer peptide. In some embodiments, the spacer peptide is uncharged. In some embodiments, the spacer peptide is a glycine-serine linker. In some embodiments, the spacer peptide contains a glycine-serine linker. In some embodiments, the spacer peptide contains or consists of about 3 to about 55 amino acids. The spacer peptide of the present invention contains or may consist of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 amino acids. In some embodiments, the spacer peptide has an amino acid length of approximately 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, or 155 amino acids (including any value or range between these values). In some embodiments, the spacer peptide contains 15 amino acids. In some embodiments, the spacer peptide contains the amino acid sequence of SEQ ID NO: 23. In some embodiments, the spacer peptide contains the amino acid sequence of SEQ ID NO: 24. In some embodiments, the spacer peptide contains the amino acid sequence of SEQ ID NO: 27. In some embodiments, the antibody-peptide fusion protein does not contain a spacer between the amyloid-reactive peptide and the antibody.

[0260] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, and a light chain containing VL with an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, wherein the light chain is linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, which does not contain a C-terminal lysine residue, and a VL with an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, wherein the antibody is linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with an amino acid sequence described in SEQ ID NOs. 51, which does not contain a C-terminal lysine residue, and a VL with an amino acid sequence described in the group consisting of SEQ ID NOs. 35, wherein the light chain is linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with an amino acid sequence described in SEQ ID NOs. 51, which does not contain a C-terminal lysine residue, and a VL with an amino acid sequence described in SEQ ID NOs. 35, wherein the antibody is linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 34, the light chain being linked to the peptide. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48 without a C-terminal lysine residue and a VL with the amino acid sequence of SEQ ID NO: 34, the antibody being linked to the peptide.

[0261] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs: 44-63, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs: 44-63, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs: 44-63, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody containing a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing a VH having the amino acid sequence of SEQ ID NO: 48, the heavy chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 2.

[0262] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL having an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, and the light chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL having an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL having an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 2. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL having an amino acid sequence of SEQ ID NO. 35, and the light chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL having an amino acid sequence of SEQ ID NO. 35, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 35, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 34, and the light chain is linked to a peptide containing any of the amino acid sequences in Table 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 34, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 34, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0263] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 35, wherein the light chain is linked to a peptide containing any of the amino acid sequences in Table 1.

[0264] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 35, wherein the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0265] In some embodiments, the antibody-peptide fusion protein comprises a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 35. The material contains a tethered antibody, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0266] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 34, wherein the light chain is linked to a peptide containing one of the amino acid sequences in Table 1.

[0267] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 34, wherein the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0268] In some embodiments, the antibody-peptide fusion protein comprises a humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 34, wherein the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the light chain at its N-terminus.

[0269] C. Humanized antibodies and antibody-peptide fusion proteins containing humanized antibodies In some embodiments, the humanized antibody or antibody-peptide fusion protein comprising a humanized antibody of this disclosure comprises an Fc region. In some embodiments, Fc is the Fc of an IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the humanized antibody or antibody-peptide fusion protein comprising a humanized antibody promotes Fc-mediated antibody effector function. In some embodiments, the humanized antibody or antibody-peptide fusion protein comprising a humanized antibody promotes antibody-dependent cell phagocytosis.

[0270] In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of less than approximately 100, 10, 1, 0.1, or 0.01 μM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of approximately 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 humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of less than 500, 100, 10, or 1 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of approximately 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 250, 500, 750, 1000, 2000, or less than 2200 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of approximately 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 humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of approximately 40-50 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of 40-50 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of less than 50 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils with a dissociation constant (Kd) of less than the Kd at which c11-1F4 binds to human amyloid fibrils.

[0271] In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) that is less than about 0.01, 0.1, or 1 μM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) that is 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, or 10 μM (including any value or range between these values). In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) that is less than about 1, 10, 100, or 1000 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) 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 between these values). In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) that is about 17 nM, 7 nM, 16 nM, 75 nM, or 95 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) that is less than about 10 nM, 20 nM, 80 nM or 100 nM. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to human amyloid fibrils at an antibody half-maximal binding concentration (EC 50 ) that is less than the EC 50 ) of c11-1F4 that binds to human amyloid fibrils.

[0272] Dissociation constant and EC 50Methods for calculating the dissociation constant are known in the art and include, for example, surface plasmon resonance and EuLISA (see, for example, Examples, Table 7, and Figures 13A-13G). In some embodiments, the dissociation constant is determined, for example, by measuring the binding to the Len(1-22) monomer peptide using surface plasmon resonance. In some embodiments, EC 50 This is determined using EuLISA. In some embodiments, EC 50 The binding levels to rVλ6Wil fibrils, Per125 wtATTR extract, Ken ATTR extract, SHI ALλ liver extract, or TAL ALκ liver extract are determined using EuLISA.

[0273] In some embodiments, a humanized antibody or antibody-peptide fusion protein is conjugated with a detectable label. In some embodiments, the detectable label is a radionuclide (e.g., I- 125 , I- 123 , I- 131 , Zr- 89 ,Tc- 99m , Cu- 64 , Br- 76 , F- 18 ), selected from the group consisting of enzymes (horseradish peroxidase), biotin, and fluorophores. Any known means in the art for detectably labeling proteins can be used and / or adapted for use with the methods described herein. For example, humanized antibodies or antibody-peptide fusion proteins can be radiolabeled with radioisotopes or labeled with fluorescent or chemiluminescent tags. Exemplary radioisotopes include, for example, 18 F, 111 In, 99m Tc, and 123 I and 125I is an example. These and other radioisotopes can be conjugated to humanized antibodies or antibody-peptide fusion proteins using well-known chemistry, which may or may not involve the use of chelating agents such as DTPA or DOTA covalently bound to the light chain protein of the humanized antibody or antibody-peptide fusion protein. Exemplary fluorescent or chemiluminescent tags include fluorescein, Texas Red, rhodamine, Alexa dye, and luciferase, which can be conjugated to humanized antibodies or antibody-peptide fusion proteins by reaction with lysine, cysteine, glutamic acid, and aspartic acid side chains. In one example embodiment, the label is applied to a fluorescent microplate using excitation and emission wavelengths appropriate to the tag used. It is detected using a reader or a fluorometer. Radioactive labels can be detected, for example, by using a gamma counter or scintillation counter depending on the type of radioactive emission, and by using an energy window suitable for the precise detection of a specific radionuclide. However, any other suitable technique for detecting radioisotopes can also be used to detect the labels. In some embodiments, the detectable labels are 125 It is I.

[0274] In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to rVλ6Wil fibril, Per125 wtATTR extract, KEN hATTR extract, SHI ALλ liver extract, and / or TAL ALκ liver extract. In some embodiments, the humanized antibody or antibody-peptide fusion protein described herein binds to amyloid deposits or amyloid fibril. In some embodiments, the humanized antibody or antibody-peptide fusion protein binds to one or more amyloidogenic peptides of amyloid. In some embodiments, the amyloid bound by the humanized antibody or antibody-peptide fusion protein includes amyloidogenic λ6 variable domain protein (Vλ6Wil) or amyloidogenic immunoglobulin light chain (AL), Aβ(1-40) amyloidoid fibril or amyloidogenic Aβ precursor protein, or serum amyloid protein A (AA). In other embodiments, the amyloid conjugated by a humanized antibody or antibody-peptide fusion protein 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 A variant (AFib), cystatin variant (ACys), calcitonin (ACal), lactoadherin (AMed), islet amyloid polypeptide (AIAPP), prolactin (APro), insulin (AIns), prion protein (APrP), α-synuclein (AαSyn), tau (ATau), atrial natriuretic factor (AANF), or IAAP, ALκ4, Alλ1, or other amyloidogenic peptides. The amyloidogenic peptide conjugated by a humanized antibody or antibody-peptide fusion protein may be a protein, protein fragment, or protein domain. In some embodiments, the amyloid deposit or amyloid fibrils contain recombinant amyloidogenic proteins. In some embodiments, amyloid is part of the pathology of the disease.

[0275] In some embodiments, the binding of a humanized antibody or antibody-peptide fusion protein to human amyloid promotes phagocytosis of human amyloid fibrils. In some embodiments, the humanized antibody or antibody-peptide fusion protein opsonizes human amyloid fibrils. In some embodiments, the humanized antibody or antibody-peptide fusion protein opsonizes rVλ6Wil fibrils. In some embodiments, contact of human amyloid fibrils with the humanized antibody or antibody-peptide fusion protein of this disclosure in the presence of macrophages promotes the uptake of human amyloid fibrils by macrophages. In some embodiments, contact of human amyloid fibrils with the humanized antibody or antibody-peptide fusion protein of this disclosure in the presence of macrophages promotes the opsonization of human amyloid fibrils. In some embodiments, the binding of a humanized antibody or antibody-peptide fusion protein to human amyloid promotes phagocytosis of human amyloid fibrils to a degree equivalent to or greater than that of a control antibody (e.g., mIgp5 and / or c11-1F4). In some embodiments, a humanized antibody or an antibody-peptide fusion protein containing a humanized antibody promotes antibody-dependent cellular phagocytosis.

[0276] Pharmaceutical compositions comprising any of the modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins described herein are also provided herein. In some embodiments The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0277] IV. Diagnostic and Detection Methods In certain exemplary embodiments, modified immunoglobulins, humanized antibodies, and antibody-peptide fusion proteins may be labeled with various drugs to enable their detection in in vivo and in vitro assays, for example, after purification of the fusion peptide. 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 This includes, for example, enzymes (horseradish peroxidase), biotin, and fluorophores. Any known means in the art for detectably labeling proteins can be used and / or adapted for use with the methods described herein. For example, Ig antibodies or fragments thereof, and / or amyloid-reactive peptides can be radiolabeled with radioisotopes, or labeled with fluorescent or chemiluminescent tags. Exemplary radioisotopes include, for example, 18 F, 111 In, 99m Tc, and 123 I and 125 I is one example. These and other radioactive isotopes can be conjugated to isolated immunoglobulin light chains using well-known chemistry, which may or may not involve the use of chelating agents such as DTPA or DOTA covalently bound to the light chain protein of an Ig antibody. Exemplary fluorescent or chemiluminescent tags include fluorescein, Texas Red, rhodamine, Alexa dye, 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 fluorophotometer with excitation and emission wavelengths suitable for the tag used. Radioactive labels can be detected, for example, using a gamma counter or scintillation counter depending on the type of radioactive emission, by using an energy window suitable for the precise detection of a particular radionuclide. However, any other suitable techniques for detecting radioactive isotopes can also be used to detect labels.

[0278] With respect to amyloidosis, for example, such labels can be used to diagnose the presence of amyloid, determine the amyloid protein load, monitor the ability of modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins to bind to amyloid in a particular subject, monitor the progression of amyloidosis, and / or monitor the subject's response to amyloid treatment (including treatments involving the administration of modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins to the subject). For example, modified immunoglobulins containing amyloid-reactive peptides, humanized antibodies, or antibody-peptide fusion proteins are labeled with the detectable labels described herein and then administered to a subject suffering from or suspected of suffering from an amyloid disorder (e.g., amyloidosis, monoclonal immunoglobulinemia of unspecified significance (MGUS), multiple myeloma (MM), or related plasma cell disorders). The subject can then be imaged to detect the presence of the detectably labeled immunoglobulin, humanized antibody, or antibody-peptide fusion protein.

[0279] In certain exemplary embodiments, signals from detectably labeled modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins can be quantified, thereby providing an indicator of the level of amyloid deposition in a subject. For example, the signal intensity can be compared to a standard signal threshold, where a threshold above indicates the presence of amyloidosis, and a threshold below indicates the absence or low level of amyloidosis. A subject may be diagnosed with amyloidosis, in which case treatment such as chemotherapy, corticosteroids (lenalidomide or thalidomide), and / or bortezomib (Velcade) may be administered. Additionally or alternatively, the modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins described herein may be used as described herein. These can be administered to a subject to treat it. In certain exemplary embodiments, subjects may be stratified into one or more groups, such as low amyloid loading, medium amyloid loading, or high amyloid loading, and then treated accordingly. To monitor the progress of treatment, subjects may be readjusted with detectably labeled modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins, and thus their amyloid loading may be reassessed.

[0280] V. Treatment Methods A. Method using modified immunoglobulins In certain exemplary embodiments, a method for treating a subject having amyloidosis is provided herein. For example, the subject is administered an effective amount of a modified immunoglobulin described herein, thereby treating the subject or enabling imaging of amyloid deposits. In certain exemplary embodiments, a method for clearing amyloid deposits in a subject is provided. This method includes, for example, selecting a subject having amyloidosis and administering an effective amount of a modified immunoglobulin described herein to the subject. Examples of modified immunoglobulins include amyloid-reactive peptides that bind to amyloid deposits, ligated to an Ig antibody or fragment via the N-terminus of the light chain protein or the C-terminus of the heavy chain of an Ig antibody or fragment thereof. Administration of the amyloid-reactive Ig fusion peptide results in amyloid clearance, thereby treating the subject.

[0281] In some embodiments, modified immunoglobulins bind to amyloid in an individual. In some embodiments, amyloid deposits may contribute to the pathology of the disease. In other embodiments, amyloid deposits may indicate amyloidosis or amyloid-related disease in an individual. In some embodiments, modified immunoglobulins bind to amyloid in an individual having amyloidosis. In some embodiments, amyloidosis is localized to a specific tissue or organ system, such as the liver, heart, or central nervous system. In other embodiments, amyloidosis is systemic amyloidosis. In some embodiments, amyloidosis is familial amyloidosis. In other embodiments, amyloidosis is sporadic amyloidosis. In some embodiments, amyloidosis or amyloid-related disease is AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, ALect2 amyloidosis, as well as IAPP amyloidosis in type II diabetes, Alzheimer's disease, Down syndrome, hereditary cerebral hemorrhage with Dutch type amyloidosis, cerebral β-amyloid angiopathy, cavernous encephalopathy, thyroid tumors, Parkinson's disease, Lewy body dementia, tauopathy, Huntington's disease, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, age-related pituitary disorders, iatrogenic syndromes, cavernous encephalopathy, reactive chronic inflammation, thyroid tumors, myeloma, or other forms of cancer. In some embodiments, modified immunoglobulins bind to amyloid associated with normal aging. In other embodiments, modified immunoglobulins are used in the diagnosis, treatment, or prognosis of amyloidosis or amyloid-related disease in a subject.

[0282] In certain exemplary embodiments, methods are provided for both the diagnosis and treatment of a subject suffering from amyloidosis. Such a method includes administering to the subject a detectably labeled modified immunoglobulin containing an amyloid-reactive peptide, and determining, based on the administration of the labeled modified immunoglobulin, that the subject suffers from amyloidosis. An effective amount of an amyloid therapeutic agent can then be administered to the subject. For example, one or more modified immunoglobulins containing an effective amount of amyloid-reactive peptide can be administered.

[0283] In some embodiments, the subject is a mammal such as a primate, cattle, rodent, or pig. In some embodiments, the subject is a human.

[0284] B. Methods using humanized antibodies or antibody-peptide fusion proteins Methods for treating subjects with amyloid-related disorders are also provided herein, comprising administering to the subject an effective amount of the humanized antibody or antibody-peptide fusion protein of this disclosure.

[0285] In some embodiments, amyloid-related disorders are selected from the group consisting of AL, AH, Aβ2M, ATTR, transthyretin, AA, AApoAI, AApoAII, AGel, Alys, ALEct2, AFib, ACys, ACa, AMed, AIAPP, APro, AIns, APrP, α-synuclein, au, or Aβ amyloidosis. In some embodiments, amyloidosis or amyloid-related disorders include AA amyloidosis, AL amyloidosis, AH amyloidosis, Aβ amyloidosis, ATTR amyloidosis, ALect2 amyloidosis, as well as IAPP amyloidosis in type II diabetes, Alzheimer's disease, Down syndrome, hereditary cerebral hemorrhage with Dutch amyloidosis, cerebral β-amyloid angiopathy, cavernous encephalopathy, thyroid tumors, Parkinson's disease, Lewy body dementia, tauopathy, Huntington's disease, senile systemic amyloidosis, familial hemodialysis, senile systemic aging, age-related pituitary disorders, iatrogenic syndromes, cavernous encephalopathy, reactive chronic inflammation, thyroid tumors, myeloma, or other forms of cancer. In some embodiments, amyloid-related disorders are systemic amyloidosis. In some embodiments, humanized antibodies or antibody-peptide fusion proteins bind to amyloid associated with normal aging. In other embodiments, humanized antibodies or antibody-peptide fusion proteins are used in the diagnosis, treatment, or prognosis of amyloidosis or amyloid-related diseases in subjects.

[0286] Methods for targeting amyloid deposits for clearance are also provided herein. In some embodiments, the method involves contacting the amyloid deposits with a humanized antibody or antibody-peptide fusion protein of the Disclosure. In some embodiments, the amyloid deposits are removed. In some embodiments, the amyloid deposits are cleared. In some embodiments, the amyloid deposits are opsonized by the humanized antibody or antibody-peptide fusion protein. In some embodiments, binding of the humanized antibody or antibody-peptide fusion protein to human amyloid fibrils facilitates phagocytosis of human amyloid fibrils and removal of amyloid deposits. In some embodiments, the humanized antibody or antibody-peptide fusion protein opsonizes human amyloid fibrils, thereby removing amyloid deposits. In some embodiments, the humanized antibody or antibody-peptide fusion protein opsonizes rVλ6Wil fibrils. In some embodiments, the binding of a humanized antibody or antibody-peptide fusion protein to human amyloid fibrils promotes phagocytosis and / or opsonization of human amyloid fibrils to a degree equivalent to or greater than that of a control antibody (e.g., mIgp5 and / or c11-1F4).

[0287] In some embodiments, a method for treating amyloid-related disorders is provided herein, comprising administering a modified immunoglobulin or antibody-peptide fusion protein conjugated with a detectable label, detecting the label, and, if a signal is detected, administering amyloidosis treatment to the subject. In some embodiments, the detectable label is a radiolabel. In some embodiments, the detectable label is I 125 , Tc 99The 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 (e.g., biotin), which can be detected by binding to a specific homologous detectable moiety (e.g., labeled avidin). In some embodiments, amyloid deposits are identified in the liver, spleen, or blood of the subject. In some embodiments, the treatment of amyloidosis comprises a modified immunoglobulin or antibody-peptide fusion protein provided herein.

[0288] Also provided herein are methods for identifying amyloid deposits in a subject, comprising administering a modified immunoglobulin or antibody-peptide fusion protein, wherein the modified immunoglobulin or antibody-peptide fusion protein is conjugated to a detectable label. In some embodiments, the method comprises detecting a signal from the modified immunoglobulin or antibody-peptide fusion protein. In some embodiments, the detectable label is a radiolabel. In some embodiments, the detectable label is I 125 , Tc 99 The 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 (e.g., biotin), which can be detected by binding to a specific homologous detectable moiety (e.g., labeled avidin). In some embodiments, amyloid deposits are identified in the liver, spleen, or blood of the subject.

[0289] In some embodiments, a method for detecting a ligand is provided herein, comprising contacting the ligand with a modified immunoglobulin or antibody-peptide fusion conjugated with a detectable label, and determining a signal from the detectable label. In some embodiments, the detectable label is an emitting label. In some embodiments, the detectable label is I125 , Tc 99 The 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 (e.g., biotin), which can be detected by binding to a specific homologous detectable moiety (e.g., labeled avidin). In some embodiments, the contact is in vitro. In some embodiments, the contact is in vivo.

[0290] VI. Methods for producing nucleic acids, vectors, host cells, and antibodies A. Nucleic acids encoding modified immunoglobulins or antibody-peptide fusion proteins Nucleic acids encoding modified immunoglobulins are also provided herein. In some embodiments, the nucleic acid encodes one of the modified immunoglobulins described herein.

[0291] In some embodiments, the nucleic acid encodes a modified immunoglobulin containing an antibody comprising the VH and / or VL of antibody 11-1F4, and the antibody is linked to a peptide. In some embodiments, the nucleic acid encodes an antibody comprising the heavy chain and / or light chain of antibody 11-1F4, and the antibody is linked to a peptide.

[0292] In a particular embodiment, the nucleic acid encodes a modified immunoglobulin comprising an antibody comprising (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 17, (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 18, and (c) VH comprising CDR-H3 comprising the amino acid sequence of SEQ ID NO: 19, the antibody being linked to a peptide.

[0293] In a particular embodiment, the nucleic acid encodes a modified immunoglobulin comprising an antibody comprising (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 21, and (c) VL comprising CDR-L3 comprising the amino acid sequence of SEQ ID NO: 22, the antibody being linked to the peptide.

[0294] In one embodiment, the nucleic acid encodes a modified immunoglobulin comprising an antibody containing VL, which contains the amino acid sequence of SEQ ID NO: 16, and VH, which contains the amino acid sequence of SEQ ID NO: 15, and the antibody is linked to the peptide.

[0295] In another embodiment, the nucleic acid is CDR-H1 containing the amino acid sequence of SEQ ID NO: 17, CDR-H2 containing the amino acid sequence of SEQ ID NO: 18, and CDR- containing the amino acid sequence of SEQ ID NO: 19. The modified immunoglobulin encodes an antibody containing VH containing H3, and VL containing CDR-L1 containing the amino acid sequence of SEQ ID NO: 20, CDR-L2 containing the amino acid sequence of SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence of SEQ ID NO: 22, wherein the antibody is linked to a peptide. In some embodiments, the modified immunoglobulin includes an antibody linked to an amyloid-reactive peptide containing one of the amino acid sequences listed in Table 1. In some embodiments, the modified immunoglobulin includes an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin includes an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 2.

[0296] In another embodiment, the nucleic acid encodes a modified immunoglobulin comprising antibodies containing VH CDR1, VH CDR2, and VH CDR3 of VH having the sequence described in SEQ ID NO: 15, and VL CDR1, VL CDR2, and VL CDR3 of VL having the sequence described in SEQ ID NO: 16, wherein the antibodies are linked to peptides. In some embodiments, the modified immunoglobulin comprises antibodies linked to amyloid-reactive peptides containing any of the amino acid sequences listed in Table 1. In some embodiments, the modified immunoglobulin comprises antibodies linked to amyloid-reactive peptides containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modified immunoglobulin comprises antibodies linked to amyloid-reactive peptides containing the amino acid sequence of SEQ ID NO: 2.

[0297] In some embodiments, the nucleic acid encodes a modified immunoglobulin comprising an antibody comprising an antibody heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide. In some embodiments, the modified immunoglobulin comprises an antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 15 without a C-terminal lysine residue and a light chain containing the amino acid sequence of SEQ ID NO: 16, the antibody being linked to a peptide.

[0298] In another embodiment, the nucleic acid encodes a modified immunoglobulin comprising an antibody comprising VH from any of the above-provided embodiments and VL from any of the above-provided embodiments, wherein the antibody is linked to a peptide.

[0299] In some embodiments, the nucleic acid encodes an antibody linked to an amyloid-reactive peptide. In some embodiments, the nucleic acid encodes an antibody linked to an amyloid-reactive peptide containing one of the amino acid sequences listed in Table 1. In some embodiments, the nucleic acid encodes an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody containing a light chain linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the N-terminus of the antibody light chain or the C-terminus of the heavy chain. In some embodiments, the nucleic acid encodes an antibody also containing a spacer amino acid sequence between the peptide and the N-terminus of the antibody light chain or the C-terminus of the heavy chain. In some embodiments, the peptide is linked to the N-terminus of the antibody light chain.

[0300] In some embodiments, the nucleic acid encodes a modified immunoglobulin comprising an antibody comprising a heavy chain comprising VH containing the amino acid sequence of SEQ ID NO: 15, linked to a peptide comprising one of the amino acid sequences of Table 1. In some embodiments, the nucleic acid encodes an antibody comprising a heavy chain comprising VH containing the amino acid sequence of SEQ ID NO: 15, linked to a peptide comprising the amino acid sequence of SEQ ID NO:2.

[0301] In some embodiments, the nucleic acid encodes a modified immunoglobulin comprising an antibody comprising a light chain comprising a VL containing the amino acid sequence of SEQ ID NO: 16 linked to a peptide containing one of the amino acid sequences of Table 1. In some embodiments, the nucleic acid comprises the amino acid sequence of SEQ ID NO: 1 The nucleic acid encodes an antibody comprising a light chain containing a VL containing the amino acid sequence of SEQ ID NO: 16 linked to a peptide. In some embodiments, the nucleic acid encodes an antibody comprising a light chain containing a VL containing the amino acid sequence of SEQ ID NO: 16 linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0302] In some embodiments, the nucleic acid encodes a modified immunoglobulin containing an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide containing one of the amino acid sequences in Table 1.

[0303] In some embodiments, the nucleic acid encodes a modified immunoglobulin comprising an antibody including a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1.

[0304] In some embodiments, the nucleic acid encodes a modified immunoglobulin comprising an antibody including a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0305] Furthermore, nucleic acids (or more) encoding antibody-peptide fusion proteins are also provided herein. In some embodiments, the nucleic acid encodes one of the antibody-peptide fusion proteins described herein.

[0306] In a particular embodiment, the nucleic acid encodes an antibody-peptide fusion protein containing an antibody, the antibody comprising VH including (a) CDR-H1 containing the amino acid sequence of SEQ ID NO: 17, (b) CDR-H2 containing the amino acid sequence of SEQ ID NO: 18, and (c) CDR-H3 containing the amino acid sequence of SEQ ID NO: 19, and the antibody is linked to the peptide.

[0307] In a particular embodiment, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody, the antibody comprising VL comprising (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20, (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 21, and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 22, and the antibody is linked to the peptide.

[0308] In one embodiment, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody containing VL, which contains the amino acid sequence of SEQ ID NO: 16, and VH, which contains the amino acid sequence of SEQ ID NO: 15, wherein the antibody is linked to the peptide.

[0309] In another embodiment, the nucleic acid encodes an antibody-peptide fusion protein containing an antibody, wherein the antibody comprises VH, which includes CDR-H1 containing the amino acid sequence of SEQ ID NO: 17, CDR-H2 containing the amino acid sequence of SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence of SEQ ID NO: 19; and VL, which includes CDR-L1 containing the amino acid sequence of SEQ ID NO: 20, CDR-L2 containing the amino acid sequence of SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence of SEQ ID NO: 22, and the antibody is linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing an antibody linked to an amyloid-reactive peptide containing one of the amino acid sequences listed in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing an antibody linked to an amyloid-reactive peptide containing the amino acid sequence of SEQ ID NO: 2.

[0310] In another embodiment, the nucleic acid encodes an antibody-peptide fusion protein containing an antibody, and the antibody is VH CDR1, VH CDR2, and VH having the sequence described in SEQ ID NO: 15. The nucleic acid comprises CDR3 and VL CDR1, VL CDR2, and VL having the sequence described in SEQ ID NO: 16, and the antibody is linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide having one of the amino acid sequences listed in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide having the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide having the amino acid sequence of SEQ ID NO: 2.

[0311] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody having a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, the light chain being linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 without a C-terminal lysine residue and a VL having the amino acid sequence of SEQ ID NO: 16, the antibody being linked to the peptide.

[0312] In another embodiment, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide comprising one of the amino acid sequences listed in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody linked to an amyloid-reactive peptide comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the peptide is linked to the N-terminus of the light chain or the C-terminus of the heavy chain of the antibody. In some embodiments, the antibody also comprises a spacer amino acid sequence between the peptide and the N-terminus of the light chain or the C-terminus of the heavy chain.

[0313] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 15, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0314] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody comprising a light chain comprising a VL containing the amino acid sequence of SEQ ID NO: 16, and the light chain is linked to a peptide comprising one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody comprising a light chain comprising a VL containing the amino acid sequence of SEQ ID NO: 16, and the light chain is linked to a peptide comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody comprising a light chain comprising a VL containing the amino acid sequence of SEQ ID NO: 16, and the light chain is linked to a peptide comprising the amino acid sequence of SEQ ID NO: 2.

[0315] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody having a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, wherein the light chain is linked to a peptide having one of the amino acid sequences in Table 1.

[0316] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody having a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 1.

[0317] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising an antibody having a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 15 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 16, the light chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 2.

[0318] B. Nucleic acids encoding humanized antibodies or antibody-peptide fusion proteins containing humanized antibodies Furthermore, nucleic acids (or multiple nucleic acids) encoding humanized antibodies or antibody-peptide fusion proteins are also provided herein. A humanized antibody or antibody-peptide fusion protein may be any of the humanized antibodies or antibody-peptide fusion proteins described herein.

[0319] In some embodiments, the nucleic acid encodes a humanized antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), where the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the nucleic acid encodes a humanized antibody comprising 1, 2, 3, 4, 5, or 6 CDRs of the antibody, as shown in Table 3. In some embodiments, the nucleic acid encodes a humanized antibody comprising CDR-H1, CDR-H2, and CDR-H3, each containing the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequence described in SEQ ID NO: 15, and CDR-L1, CDR-L2, and CDR-L3, each containing the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequence described in SEQ ID NO: 16.

[0320] In some embodiments, the nucleic acid encodes a humanized antibody, and the humanized antibody includes one or more CDR substitutions. In some embodiments, the humanized antibody includes VL, which includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 64-70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22; and VH, which includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 19. In some embodiments, the nucleic acid encodes a humanized antibody, and the humanized antibody includes VL, which includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs. 20, CDR-L2 containing the amino acid sequence described in SEQ ID NOs. 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs. 22; and VH, which includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs. 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs. 71-81, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs. 19.

[0321] In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing one or more amino acid substitutions at one or more positions of the VL compared to a VL containing the amino acid sequence described in SEQ ID NO: 32, where the amino acid positions are sequentially numbered starting from the N-terminus of SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 33 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 34 compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. The nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 41, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 42, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 51, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 90, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing an amino acid substitution at position 92, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions, compared to a VL containing the amino acid sequence described in SEQ ID NO: 32. In some embodiments, the nucleic acid encodes a humanized antibody, comprising a VL containing 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions compared to VL1, as shown in Table 6A.

[0322] In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising one or more amino acid substitutions at one or more positions of VH compared to VH comprising the amino acid sequence described in SEQ ID NO: 43, where the amino acid positions are numbered starting from the N-terminus of SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising an amino acid substitution at position 37 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising an amino acid substitution at position 48 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising an amino acid substitution at position 67 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising an amino acid substitution at position 68 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising an amino acid substitution at position 71 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH comprising an amino acid substitution at position 76 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH with an amino acid substitution at position 78 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH with an amino acid substitution at position 79 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH with an amino acid substitution at position 80 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH with an amino acid substitution at position 92 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH with an amino acid substitution at position 96 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH with an amino acid substitution at position 97 compared to VH comprising the amino acid sequence described in SEQ ID NO: 43.In some embodiments, the nucleic acid encodes a humanized antibody comprising VH containing 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions compared to VH containing the amino acid sequence described in SEQ ID NO: 43. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH containing 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions compared to VH1, as shown in Table 6B.

[0323] In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing one or more amino acid residues at one or more positions of the VL, where the amino acid positions are numbered starting from the N-terminus of the VL. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing Ser, Gln, Glu, His, or Ala at position 33. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing Ala or Val at position 34. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing Tyr at position 41. Some embodiments In some embodiments, the nucleic acid encodes a humanized antibody containing a VL with Leu at position 42. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL with Leu at position 51. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL with Leu at position 90. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL with Phe at position 92.

[0324] In some embodiments, the nucleic acid encodes a humanized antibody containing VH with one or more amino acid residues at one or more positions of VH, where the amino acid positions are numbered starting from the N-terminus of VH. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 37. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 48. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 67. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Ser at position 68. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Lys at position 71. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Ser at position 76. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 78. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 79. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Phe at position 80. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Thr at position 92. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 96. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Thr at position 97.

[0325] In some embodiments, the nucleic acid encodes a humanized antibody containing a VL that includes one or more amino acid residues at one or more positions of the VL, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL that includes Tyr at position 36. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL that includes Leu at position 37. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL that includes Leu at position 46. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL that includes Leu at position 85. In some embodiments, the nucleic acid encodes a humanized antibody containing a VL that includes Phe at position 87.

[0326] In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing one or more amino acid residues at one or more positions of the VL, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing Tyr at position 36 and Leu at position 37. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87. In some embodiments, the nucleic acid encodes a humanized antibody comprising a VL containing Leu at position 46 and Phe at position 87.

[0327] In some embodiments, the nucleic acid encodes a humanized antibody containing VH with one or more amino acid residues at one or more positions of VH, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 37. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 48. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 67. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Ser at position 68. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Lys at position 71. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Ser at position 76. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 78. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 79. In some embodiments, the nucleic acid is at position Position 80 encodes a humanized antibody containing VH with Phe. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Thr at position 89. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 93. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Thr at position 94.

[0328] In some embodiments, the nucleic acid encodes a humanized antibody containing VH with one or more amino acid residues at one or more positions of VH, where the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 37 and Leu at position 48. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 67, Ser at position 68, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 37, Leu at position 48, Leu at position 67, and Ser at position 68. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Val at position 37, Leu at position 48, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH containing Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH containing Lys at position 71, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH containing Lys at position 71, Ser at position 76, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VH containing Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody containing VH with Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94.

[0329] In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and VH containing Val at position 37, Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Thr at position 89, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Leu at position 48, Ser at position 96, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Leu at position 48, Leu at position 67, Ser at position 68, Lys at position 71, Ser at position 76, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Lys at position 71, Ser at position 76, Val at position 93, and Thr at position 94. In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing Leu at position 46 and Phe at position 87, and VH containing Lys at position 71, Val at position 78, Leu at position 79, Val at position 93, and Thr at position 94.

[0330] In some embodiments, the nucleic acid encodes a humanized antibody containing the amino acid sequence of VL shown in Table 6A. In some embodiments, the nucleic acid encodes a humanized antibody containing 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 6A. In some embodiments, the nucleic acid encodes a humanized antibody containing VL containing the amino acid sequence described in the group consisting of SEQ ID NOs.33 to 42.

[0331] In some embodiments, the nucleic acid encodes a humanized antibody containing the amino acid sequence of VH shown in Table 6B. In some embodiments, the nucleic acid encodes a humanized antibody containing 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 shown in Table 6B. In some embodiments, the nucleic acid encodes a humanized antibody containing VH containing the amino acid sequence described in the group consisting of SEQ ID NOs.44 to 63.

[0332] In some embodiments, the nucleic acid encodes a humanized antibody comprising VL of VL4 shown in Table 6A and VH of VH9 shown in Table 6B. In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing the amino acid sequence described in SEQ ID NO: 35 and VH containing the amino acid sequence described in SEQ ID NO: 51.

[0333] In some embodiments, the nucleic acid encodes a humanized antibody comprising VL of VL3 shown in Table 6A and VH of VH6 shown in Table 6B. In some embodiments, the nucleic acid encodes a humanized antibody comprising VL containing the amino acid sequence described in SEQ ID NO: 34 and VH containing the amino acid sequence described in SEQ ID NO: 48.

[0334] In some embodiments, the nucleic acid encodes a humanized antibody comprising the VL of VL4 shown in Table 6A and the VH of VH10 shown in Table 6B. In some embodiments, the nucleic acid encodes a humanized antibody comprising the VL containing the amino acid sequence described in SEQ ID NO: 35 and the VH containing the amino acid sequence described in SEQ ID NO: 52. In some embodiments, the nucleic acid encodes a humanized antibody comprising the VL of VL4 shown in Table 6A and the VH of VH8 shown in Table 6B. In some embodiments, the nucleic acid encodes a humanized antibody comprising the VL containing the amino acid sequence described in SEQ ID NO: 35 and the VH containing the amino acid sequence described in SEQ ID NO: 50. In some embodiments, the nucleic acid encodes a humanized antibody comprising the VL of VL4 shown in Table 6A and the VH of VH7 shown in Table 6B. In some embodiments, the VL contains the amino acid sequence described in SEQ ID NO: 35, and the VH contains the amino acid sequence described in SEQ ID NO: 69.

[0335] Also provided herein are nucleic acids (may include multiple) encoding antibody-peptide fusion proteins, including humanized antibodies of the present disclosure. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein, including one of the humanized antibodies of the present disclosure.

[0336] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, and the humanized antibody comprises a heavy chain containing VH having an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, and a light chain containing VL having an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, the light chain being linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein, and the humanized antibody comprises a heavy chain containing VH having an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, which does not contain a C-terminal lysine residue, and a VL having an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, the antibody being linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, and the humanized antibody comprises a heavy chain containing VH having an amino acid sequence of SEQ ID NO. 51, and a light chain containing VL having an amino acid sequence of SEQ ID NO. 35, the light chain being linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51 without a C-terminal lysine residue, and VL with the amino acid sequence of SEQ ID NO: 35, and the antibody is linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48, and SEQ ID NO: 34 The nucleic acid comprises a light chain containing a VL containing the amino acid sequence of the sequence, and the light chain is linked to the peptide. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein comprising a humanized antibody, the humanized antibody comprising a heavy chain containing a VH containing the amino acid sequence of SEQ ID NO: 48 without a C-terminal lysine residue, and a VL containing the amino acids of SEQ ID NO: 34, and the antibody is linked to the peptide.

[0337] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprises a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprises a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprises a heavy chain containing VH with an amino acid sequence described in the group consisting of SEQ ID NOs. 44 to 63, and the heavy chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 2. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprises a heavy chain containing VH with an amino acid sequence of SEQ ID NO. 51, and the heavy chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51, and the heavy chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51, and the heavy chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48, and the heavy chain being linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48, and the heavy chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1.In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody containing a heavy chain containing a VH having the amino acid sequence of SEQ ID NO: 48, and the heavy chain is linked to a peptide having the amino acid sequence of SEQ ID NO: 2.

[0338] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody includes a light chain containing a VL with an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, and the light chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody includes a light chain containing a VL with an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody includes a light chain containing a VL with an amino acid sequence described in the group consisting of SEQ ID NOs. 33 to 42, and the light chain is linked to a peptide containing the amino acid sequence of SEQ ID NO. 2. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody includes a light chain containing a VL with an amino acid sequence of SEQ ID NO. 35, and the light chain is linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 35, and the light chain being linked to a peptide with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 35, and the light chain being linked to a peptide with the amino acid sequence of SEQ ID NO: 2. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 34, and the light chain being linked to a peptide containing one of the amino acid sequences in Table 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 34, and the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 1. In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a light chain containing a VL with the amino acid sequence of SEQ ID NO: 34, and the light chain being linked to a peptide containing the amino acid sequence of SEQ ID NO: 2.

[0339] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 51 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 35, the light chain being linked to a peptide containing any of the amino acid sequences in Table 1.

[0340] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 51, and a light chain containing VL having the amino acid sequence of SEQ ID NO: 35, the light chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 1.

[0341] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 51 and a light chain containing VL having the amino acid sequence of SEQ ID NO: 35, the light chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 2.

[0342] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH with the amino acid sequence of SEQ ID NO: 48 and a light chain containing VL with the amino acid sequence of SEQ ID NO: 34, the light chain being linked to a peptide containing one of the amino acid sequences in Table 1.

[0343] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 48, and a light chain containing VL having the amino acid sequence of SEQ ID NO: 34, the light chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 1.

[0344] In some embodiments, the nucleic acid encodes an antibody-peptide fusion protein containing a humanized antibody, the humanized antibody comprising a heavy chain containing VH having the amino acid sequence of SEQ ID NO: 48, and a light chain containing VL having the amino acid sequence of SEQ ID NO: 34, the light chain being linked to a peptide having the amino acid sequence of SEQ ID NO: 2.

[0345] C. Vector, host cell In some embodiments, the nucleic acids provided herein are contained in one or more vectors. For example, in some embodiments, a vector containing a heavy chain and a light chain of a modified immunoglobulin is provided herein, the light chain being ligated to a peptide. In some embodiments, the heavy chain and light chain ligated to the peptide are contained in different vectors.

[0346] In some embodiments, the vector comprises nucleic acids (may include multiple) encoding a humanized antibody or antibody-peptide fusion protein of the Disclosure.

[0347] In the case of antibody production, the heavy and light chains linked to the peptide expression vector are in the art. The expression vector can be introduced into known and suitable antibody-producing cell lines. The introduction of the expression vector can be achieved by simultaneous transfection via electroporation or by any other suitable transformation technique available in the art. The antibody-producing cell line can then be selected, grown, and the antibody purified. The purified antibody can then be analyzed by standard techniques such as SDS-PAGE.

[0348] Host cells are also provided, containing nucleic acids encoding one of the modified immunoglobulins described herein. Suitable host cells for cloning or expression of antibody-coding vectors include prokaryotic or eukaryotic cells described herein. For example, modified immunoglobulins can be produced in bacteria, particularly when glycosylation and Fc effector function are not required. For the expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Patents 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKCLo, ed., Humana Press, Totowa, NJ, 2003), pp. 245-254 (Description of antibody fragment expression in E. coli)). After expression, the peptide-linked antibody can be isolated from the bacterial cell paste in the soluble fraction and further purified.

[0349] In some embodiments, the host cell comprises a vector containing nucleic acid(s) encoding a humanized antibody or antibody-peptide fusion protein of the present disclosure.

[0350] Suitable host cells for the expression of glycosylated antibodies can also be derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant cells and insect cells. Numerous baculovirus strains have been identified and can be used in conjunction with insect cells, particularly for the transfection of Spodoptera frugiperda cells.

[0351] Plant cell cultures can also be used as host cells. See, for example, U.S. Patents 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (which describe PLANTIBODIES® technology for producing antibodies in transgenic plants).

[0352] Vertebrate cells may also be used as hosts. For example, mammalian cell lines adapted for growth in suspension may be useful. Other examples of useful mammalian host cell lines include SV40-transformed monkey kidney CVl cell line (COS-7), human embryonic kidney cell line (293 or 293 cells, e.g., Graham et al., J. Gen Virol. 36:59 (1977)), baby hamster kidney cells (BHK), mouse Sertoli cells (e.g., TM4 cells described in Mather, Biol. Reprod. 23:243-251 (1980)), monkey kidney cells (CVl), African green monkey kidney cells (VERO-76), human cervical cancer cells (HELA), canine kidney cells (MDCK), buffalo rat liver cells (BRL3A), human lung cells (W138), human liver cells (HepG2), mouse mammary tumor cells (MMT060562), and TRI cells (e.g., Mather et al., Annals). These include MRC5 cells and FS4 cells (described in NY.Acad.Sci.383:44-68 (1982)). 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 (Y0, NS0, and Sp2 / 0, etc.). For an overview of specific mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol.248 (BKCLo, ed., Humana Press, Totow). See a,NJ), pp.255-268 (2003).

[0353] D. Methods for producing antibodies or antibody-peptide fusion proteins Methods for producing the modified immunoglobulins, humanized antibodies, or antibody-peptide fusion proteins of the Disclosure are also provided herein. In some embodiments, the method comprises culturing host cells of the Disclosure under conditions suitable for the expression of a vector encoding the modified immunoglobulin, humanized antibody, or antibody-peptide fusion protein, and recovering the modified immunoglobulin, humanized antibody, or antibody-peptide fusion protein.

[0354] VII. Method for Humanizing Mouse Antibodies Methods for humanizing mouse antibodies are provided herein. In some embodiments, the mouse antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), where VL comprises CDR-L1, CDR-L2, and CDR-L3, and VH comprises CDR-H1, CDR-H2, and CDR-H3, and VH and VL comprise one or more framework residues. In some embodiments, CDR-L1 of the mouse antibody comprises the amino acid sequence described in SEQ ID NO: 20, CDR-L2 of the mouse antibody comprises the amino acid sequence described in SEQ ID NO: 21, CDR-L3 of the mouse antibody comprises the amino acid sequence described in SEQ ID NO: 22, CDR-H1 of the mouse antibody comprises the amino acid sequence described in SEQ ID NO: 17, CDR-H2 of the mouse antibody comprises the amino acid sequence described in SEQ ID NO: 18, and CDR-H3 of the mouse antibody comprises the amino acid sequence described in SEQ ID NO: 19. In some embodiments, the mouse antibody comprises one, two, three, four, five, or six CDRs of antibody 11-1F4, as shown in Table 3. In some embodiments, the mouse antibody comprises CDR-H1, CDR-H2, and CDR-H3, each comprising the amino acid sequences of VH CDR-H1, CDR-H2, and CDR-H3 having the sequence described in SEQ ID NO: 15, and CDR-L1, CDR-L2, and CDR-L3, each comprising the amino acid sequences of VL CDR-L1, CDR-L2, and CDR-L3 having the sequence described in SEQ ID NO: 16.

[0355] In some embodiments, the method for humanizing a mouse antibody includes: i. performing homology modeling to obtain a modeled structure of the mouse antibody; ii. calculating the solvent-accessible surface area of ​​framework residues in the modeled structure of the mouse antibody; iii. determining whether the framework residues are buried residues, and whether the buried residues have a solvent-accessible surface area of ​​less than approximately 15%; iv. providing human VH and VL; v. introducing CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 of the mouse antibody into human VH and VL, respectively, thereby generating transplanted antibodies; and vi. introducing a reversion mutation at the location of the transplanted antibody, wherein the location of the reversion mutation is a buried residue. In some embodiments, step vi is repeated. In some embodiments, step vi is performed 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times.

[0356] In some embodiments, the revertant mutation is located close to the CDR. In some embodiments, the revertant mutation is separated from the CDR by 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or fewer amino acid residues. In some embodiments, the revertant mutation is separated from the CDR by 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or fewer angstroms (including any range or value in between).

[0357] In some embodiments, the revertant mutation is introduced into human VL, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the revertant mutation is introduced into human VL, and the revertant mutation contains Tyr at position 36. In some embodiments, the revertant mutation is introduced into human VL, and the revertant mutation contains Leu at position 37. In some embodiments, the revertant mutation In some embodiments, the revertant mutation is introduced into the human VL and contains Leu at position 46. In some embodiments, the revertant mutation is introduced into the human VL and contains Leu at position 85. In some embodiments, the revertant mutation is introduced into the human VL and contains Phe at position 87.

[0358] In some embodiments, the revertant mutation is introduced into human VH, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Val at position 37. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Leu at position 48. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Leu at position 67. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Ser at position 68. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Lys at position 71. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Ser at position 76. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Val at position 78. In some embodiments, the revertant mutation is introduced into human VH, and the revertant mutation contains Leu at position 79. In some embodiments, the revertant mutation is introduced into human VH and the revertant mutation includes Phe at position 80. In some embodiments, the revertant mutation is introduced into human VH and the revertant mutation includes Thr at position 89. In some embodiments, the revertant mutation is introduced into human VH and the revertant mutation includes Val at position 93. In some embodiments, the revertant mutation is introduced into human VH and the revertant mutation includes Thr at position 94.

[0359] In some embodiments, the method further includes introducing one or more amino acid substitutions into the CDR of the transplanted antibody. In some embodiments, the humanized antibody includes a VL with amino acid substitutions in CDR-L1 compared to the VL of the transplanted antibody. In some embodiments, the amino acid substitutions in CDR-L1 are selected from the group consisting of amino acid substitutions at position 28 and amino acid substitutions at position 29, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitutions in CDR-L1 are selected from the group consisting of Ser, Gln, Glu, His, or Ala at position 28, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitutions in CDR-L1 are selected from the group consisting of Ala or Val at position 29, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the humanized antibody includes a VH with amino acid substitutions in CDR-H2 compared to the VH of the transplanted antibody. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of amino acid substitutions at positions 54, 55, or 64, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of Ser, Gln, Glu, Ala, or His at position 54, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of Ala or Val at position 55, and the amino acid positions are numbered according to the Kabat numbering system. In some embodiments, the amino acid substitution in CDR-H2 is selected from the group consisting of Val, Ile, Leu, or Ala at position 64, and the amino acid positions are numbered according to the Kabat numbering system.

[0360] Embodiment Embodiment 1. Modified immunoglobulin, an amyloid-reactive peptide comprising an amino acid sequence having at least 85% sequence identity with any one of the amino acid sequences described in SEQ ID NOs: 1 to 14, The modified immunoglobulin comprises an Ig antibody or a functional fragment thereof.

[0361] Embodiment 2. The amyloid-reactive peptide and the Ig antibody or its functional fragment are The modified immunoglobulin according to Embodiment 1, which is linked together at the N-terminus of the Ig light chain protein or the C-terminus of the Ig heavy chain protein.

[0362] Embodiment 3. The modified immunoglobulin according to Embodiment 1 or 2, wherein the modified immunoglobulin includes a spacer sequence between the amyloid-reactive peptide and the Ig antibody or its functional fragment.

[0363] Embodiment 4. The modified immunoglobulin according to any one of Embodiments 1 to 3, wherein the modified immunoglobulin comprises at least two amyloid-reactive peptides, and the amyloid-reactive peptides are the same peptide or different peptides.

[0364] Embodiment 5. A method for treating a subject suffering from amyloidosis, comprising administering to the subject an effective amount of a modified immunoglobulin described in any one of Embodiments 1 to 4.

[0365] Embodiment 6. A method for targeting amyloid deposits for clearance, comprising contacting the amyloid deposits with a modified immunoglobulin described in any one of Embodiments 1 to 4.

[0366] Embodiment 7. The method according to Embodiment 6, wherein targeting the amyloid deposits for clearance results in clearance of the amyloid deposits.

[0367] Embodiment 8. The method according to Embodiment 6 or 7, wherein the clearance arises from the opsonization of the amyloid deposits.

[0368] Embodiment 9. The method according to any one of Embodiments 1 to 8, wherein the amyloid-reactive peptide linked to the Ig antibody or its functional fragment is bound to one or more amyloid deposit types including AA, AL, AH, ATTR, Aβ2M, ALect2, wild type, TTR, AApoAI, AApoAII, AGel, Alys, ALect2, Afib, ACys, ACaal, AMedin, AIAPP, APro, AIns, APrP, or Aβ.

[0369] Embodiment 10. The method according to any one of Embodiments 6 to 8, wherein when the amyloid deposit is brought into contact with the amyloid-reactive peptide linked to the Ig antibody or a functional fragment thereof, the half-life of the amyloid-reactive Igp5 conjugate is increased by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or more compared to the amyloid-reactive peptide alone.

[0370] Embodiment 11. Modified immunoglobulin, Peptides and A modified immunoglobulin comprising an Ig antibody or a functional fragment thereof, wherein the peptide and the Ig antibody or functional fragment thereof are linked together at the N-terminus of an Ig light chain protein and / or the N-terminus and / or C-terminus of an Ig heavy chain protein.

[0371] Embodiment 12. The modified immunoglobulin according to Embodiment 11, wherein the modified immunoglobulin includes a spacer sequence between the peptide and the Ig antibody or its functional fragment.

[0372] Embodiment 13. The modified immunoglobulin according to any one of Embodiments 1 to 3, wherein the modified immunoglobulin comprises at least two peptides, the peptides being the same peptide or different peptides.

[0373] Embodiment 14. A method for producing modified immunoglobulins, To provide a first expression vector and a second expression vector, The first expression vector comprises a first nucleic acid sequence encoding an Ig antibody light chain or a functional fragment thereof. The second expression vector comprises a second nucleic acid sequence encoding an Ig antibody heavy chain or a functional fragment thereof. The first expression vector and / or the second expression vector comprises a third nucleic acid sequence encoding a first peptide, wherein the third nucleic acid sequence is located adjacent to the first nucleic acid sequence and / or the second nucleic acid sequence, The method comprising inserting the first and second expression vectors into a cell, wherein the expression of the first and second expression vectors in the cell results in an immunoglobulin linked to the first peptide.

[0374] Embodiment 15. The method according to Embodiment 14, wherein the first expression vector and / or the second expression vector comprises a fourth nucleic acid sequence encoding a second peptide, the fourth nucleic acid sequence being located adjacent to the first nucleic acid sequence and / or the second nucleic acid sequence.

[0375] Embodiment 16. The method according to Embodiment 15, wherein the expression of the first and second expression vectors in the cells results in immunoglobulins linked to the first and second peptides.

[0376] Embodiment 17. The method according to Embodiment 14, wherein the spacer nucleic acid sequence is located between the third nucleic acid sequence and the first nucleic acid sequence and / or the second first nucleic acid sequence.

[0377] Embodiment 18. The method according to Embodiment 14, wherein the first peptide comprises an amino acid sequence having at least 85% sequence identity with any one of the amino acids described in SEQ ID NOs: 1 to 14.

[0378] Embodiment 19. Modified immunoglobulin produced by the method described in any of Embodiments 14 to 19.

[0379] Embodiment 20. A method for treating a subject suffering from or suspected of suffering from an amyloid system disease, Whether the subject has amyloid deposits, The modified immunoglobulin described in any of Embodiments 1 to 4 is to be detectably labeled, The subject is administered the modified immunoglobulin, The determination is made by determining whether the signal associated with the detectable marker can be detected from the target, The method comprising, if the aforementioned signal is detected, administering amyloidosis treatment to the subject.

[0380] Embodiment 21. The method according to Embodiment 20, wherein if no signal is detected, the target is monitored for the subsequent development of amyloid deposits.

[0381] Embodiment 22. The method according to Embodiment 21, further comprising determining the intensity of the signal and comparing the signal with a threshold, wherein if the intensity exceeds the threshold, it is determined that the object has amyloid deposits.

[0382] Embodiment 23. The method according to any one of Embodiments 20 to 22, wherein the amyloidosis treatment comprises administering the modified immunoglobulin described in any one of claims 1 to 4 to the subject.

[0383] Embodiment 24. The method according to Embodiment 23, wherein the administration of the modified immunoglobulin results in the clearance of the amyloid deposits in the subject.

[0384] Embodiment 25. A method for identifying amyloid deposits in a subject, comprising: detecting a modified immunoglobulin described in any of Embodiments 1 to 4; administering the modified immunoglobulin to the subject; and detecting a signal from the modified immunoglobulin.

[0385] Embodiment 26. The method according to any one of Embodiments 20 to 25, wherein the subject is determined to be amyloid-free or suffering from monoclonal immunoglobulinemia of unknown significance (MGUS), multiple myeloma (MM), or one or more related plasma cell disorders.

[0386] Embodiment 27. A method for detecting a ligand, The labeling of a modified immunoglobulin according to any one of embodiments 11 to 13, wherein the peptide of the modified immunoglobulin has a binding affinity to the ligand, The ligand is brought into contact with the modified immunoglobulin, The method comprising determining a signal from the detectable label, thereby detecting the ligand.

[0387] Embodiment 28. Modified immunoglobulin, an amyloid-reactive peptide comprising any one of the amino acid sequences described in SEQ ID NOs: 1 to 14, An antibody or an immunologically active fragment thereof, wherein the antibody comprises a heavy chain variable domain including CDRH1 as described in SEQ ID NO: 17, CDRH2 as described in SEQ ID NO: 18, and CDRH3 as described in SEQ ID NO: 19, The modified immunoglobulin comprises a fragment comprising a light chain variable domain including CDRL1 described in SEQ ID NO: 20, CDRL2 described in SEQ ID NO: 21, and CDRL3 described in SEQ ID NO: 22.

[0388] Embodiment 29. The modified immunoglobulin according to claim 28, wherein the amyloid-reactive peptide comprises SEQ ID NO: 1 or SEQ ID NO: 2.

[0389] Embodiment 30. The modified immunoglobulin according to Embodiment 28 or 29, wherein the amyloid-reactive peptide is ligated to the N-terminus of the light chain of the antibody or an immunologically active fragment thereof.

[0390] Embodiment 31. A modified immunoglobulin according to any one of Embodiments 28 to 30, wherein the amyloid-reactive peptide is linked by a linker to the N-terminus of the light chain of the antibody or an immunologically active fragment thereof.

[0391] Embodiment 32. A modified immunoglobulin according to any one of Embodiments 28 to 31, wherein the antibody or its immunologically active fragment includes an Fc region.

[0392] Embodiment 33. The modified immunoglobulin according to any one of Embodiments 28 to 32, wherein the antibody is chimeric or humanized.

[0393] Embodiment 34. The modified immunoglobulin according to any one of Embodiments 28 to 32, wherein the antibody comprises a human framework sequence.

[0394] Embodiment 35. A method for treating amyloidosis in a subject, comprising administering a modified immunoglobulin described in any one of Embodiments 28 to 34 to a subject in need thereof.

[0395] Embodiment 36. A method for producing a modified immunoglobulin according to any one of Embodiments 28 to 34, comprising transforming a host cell with one or more nucleic acids encoding the modified immunoglobulin and culturing the cell under conditions for producing the modified immunoglobulin.

[0396] Embodiment 37. The modified immunoglobulin according to any one of Embodiments 1-4, 11-13, or 28-34, wherein the modified immunoglobulin binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, Alλ1, or ATTR fibril.

[0397] Embodiment 38. A nucleic acid encoding a modified immunoglobulin as described in any one of Embodiments 1-4, 11-13, or 28-34.

[0398] Embodiment 39. A host cell containing the nucleic acid described in Embodiment 38.

[0399] Embodiment 40. The host cell according to claim 39, wherein the host cell is a CHO cell. [Examples]

[0400] The following embodiments further illustrate the present invention, but should not be construed as limiting its scope. In light of this disclosure and the general level of those skilled in the art, the following embodiments are illustrative only, and it will be understood that numerous changes, modifications, and alterations may be used without departing from the subject matter of this disclosure. The accompanying drawings are intended to be considered an integral part of the specification and description of this disclosure.

[0401] As used herein, the following abbreviations apply: eq (equivalent), M (molar), μM (micromolar), N (normal), mole (mole), mmol (millimole), μmol (micromolar), nmol (nanomole), g (gram), mg (milligram), kg (kilogram), μg (microgram), L (liter), ml (milliliters), μl (microliters), cm (centimeter), mm (millimeter), μm (micrometer), nm (nanometer), °C (degrees Celsius), h (hour), min (minute), sec (second), msec (millisecond).

[0402] Example 1. Production and use of immunoglobulins fused to amyloidophilic peptides for targeting and removal of amyloid deposits. Introduction: Immunoglobulin-peptide fusion constructs are synthetic biomolecules consisting of Ig incorporating another protein or peptide. Ig confers two major advantages to the peptide of interest: firstly, enhanced plasma half-life of amyloid-reactive peptides due to the interaction between Ig and the neonatal Fc receptor; and secondly, the ability to engage and activate effector immune cells through interactions via membrane-bound Fc receptor and complement binding. To generate fusion peptide antibodies for targeting and removing amyloid deposits (e.g., clearing), we synthesized a mouse Ig-peptide fusion construct by incorporating the synthetic a...

Claims

1. An antibody-peptide fusion protein that binds to human amyloid fibrils, comprising an antibody fused to an amyloid-reactive peptide, The amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 1 or 2. The antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), a) The VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 64, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22; and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 73, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19; b) The VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs: 64 to 70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs: 18, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs: 19; c) The VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NO: 20, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequences described in SEQ ID NOs: 71 to 81, and CDR-H3 containing the amino acid sequence described in SEQ ID NO: 19; or d) The VL includes CDR-L1 containing the amino acid sequence described in SEQ ID NOs: 64 to 70, CDR-L2 containing the amino acid sequence described in SEQ ID NOs: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NOs: 22, and the VH includes CDR-H1 containing the amino acid sequence described in SEQ ID NOs: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NOs: 71 to 81, and CDR-H3 containing the amino acid sequence described in SEQ ID NOs:

19. The aforementioned antibody-peptide fusion protein.

2. The antibody-peptide fusion protein according to claim 1, wherein the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO:

2.

3. The antibody-peptide fusion protein according to claim 1, wherein the VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 64, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 73, and CDR-H3 containing the amino acid sequence described in SEQ ID NO:

19.

4. The antibody-peptide fusion protein according to claim 3, wherein VL includes Leu at position 46 and Phe at position 87, and VH includes Leu at position 48, Ser at position 76, Val at position 78, Leu at position 79, Phe at position 80, and Thr at position 94, and the amino acid positions are numbered according to the Kabat numbering system.

5. The antibody-peptide fusion protein according to claim 4, wherein VL comprises the amino acid sequence described in SEQ ID NO: 36, and VH comprises the amino acid sequence described in SEQ ID NO:

55.

6. The antibody-peptide fusion protein according to claim 1, wherein the antibody is fused to the amyloid-reactive peptide by a linker.

7. The antibody-peptide fusion protein according to claim 1, wherein the antibody includes an Fc region.

8. The antibody-peptide fusion protein according to claim 7, wherein the Fc region is an IgG1, IgG2, IgG3, or IgG4 isotype.

9. The antibody-peptide fusion protein according to any one of claims 1 to 8, wherein the antibody binds to rVλ6Wil, ALκ4, or Alλ1 fibrils.

10. The antibody is rVλ6Wil fibril, SHI ALλ liver extract, and / or TAL An antibody-peptide fusion protein according to any one of claims 1 to 8, which binds to an ALκ liver extract.

11. The antibody-peptide fusion protein according to any one of claims 1 to 8, wherein the antibody is humanized.

12. A pharmaceutical composition comprising an antibody-peptide fusion protein according to any one of claims 1 to 8.

13. A nucleic acid encoding an antibody-peptide fusion protein according to any one of claims 1 to 8.

14. A vector comprising the nucleic acid described in claim 13.

15. A host cell comprising the vector according to claim 14.

16. A method for producing an antibody-peptide fusion protein according to any one of claims 1 to 8, comprising culturing the host cell according to claim 15 under conditions suitable for the expression of the vector encoding the antibody-peptide fusion protein, and the antibody-peptide fusion protein The method comprising recovering the chlorine.

17. The use of an antibody-peptide fusion protein according to any one of claims 1 to 8 in the manufacture of a drug, wherein the drug is a drug for the treatment of amyloid-related disorder in a subject.

18. A composition comprising an antibody-peptide fusion protein according to any one of claims 1 to 8, for treating amyloid-related disorders in a subject.

19. The use according to claim 17, wherein the amyloid-related disorder is amyloidosis.

20. The use according to claim 17, wherein the amyloid-related disorder is selected from the group consisting of AL, AH, Aβ2M, ATTR, transthyretin, AA, AApoAI, AApoAII, Agel, Allys, ALECT2, Afib, Acys, Acal, Amed, AIAPP, Apro, Ains, AprP, α-synuclein, tau, or Aβ amyloidosis.

21. The use according to claim 17, wherein the amyloid-related disorder is systemic amyloidosis.

22. The use according to claim 17, wherein the amyloid-related disorder is AL.

23. The use according to claim 17, wherein the amyloid-related disorder is ATTR.

24. The use according to claim 17, wherein the subject is a human.

25. The composition according to claim 18, wherein the amyloid-related disorder is amyloidosis.

26. The composition according to claim 18, wherein the amyloid-related disorder is selected from the group consisting of AL, AH, Aβ2M, ATTR, transthyretin, AA, AApoAI, AApoAII, AGel, ALys, ALECT2, AFib, ACYs, ACaal, AMed, AIAPP, APro, AIns, APrP, α-synuclein, tau, or Aβ amyloidosis.

27. The composition according to claim 18, wherein the amyloid-related disorder is systemic amyloidosis.

28. The composition according to claim 18, wherein the amyloid-related disorder is AL.

29. The composition according to claim 18, wherein the amyloid-related disorder is ATTR.

30. The composition according to claim 18, wherein the subject is a human.

31. Use of the pharmaceutical composition according to claim 12 in the manufacture of a drug for treating amyloid-related diseases in a subject.

32. An antibody-peptide fusion protein that binds to human amyloid fibrils, comprising an antibody fused to an amyloid-reactive peptide, The aforementioned antibody-peptide fusion protein is an antibody-peptide fusion protein for the treatment of AL amyloidosis in the target, The amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 1 or 2. The antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), The VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 64, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 73, and CDR-H3 containing the amino acid sequence described in SEQ ID NO:

19. The aforementioned antibody-peptide fusion protein.

33. The antibody-peptide fusion protein according to claim 32, wherein VL comprises the amino acid sequence described in SEQ ID NO: 36, and VH comprises the amino acid sequence described in SEQ ID NO:

55.

34. The antibody-peptide fusion protein according to any one of claims 32 or 33, wherein the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO:

2.

35. An antibody-peptide fusion protein that binds to human amyloid fibrils, comprising an antibody fused to an amyloid-reactive peptide, The aforementioned antibody-peptide fusion protein is an antibody-peptide fusion protein for the treatment of ATTR amyloidosis in the target, The amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 1 or 2. The antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), The VL comprises CDR-L1 containing the amino acid sequence described in SEQ ID NO: 64, CDR-L2 containing the amino acid sequence described in SEQ ID NO: 21, and CDR-L3 containing the amino acid sequence described in SEQ ID NO: 22, and the VH comprises CDR-H1 containing the amino acid sequence described in SEQ ID NO: 17, CDR-H2 containing the amino acid sequence described in SEQ ID NO: 73, and CDR-H3 containing the amino acid sequence described in SEQ ID NO:

19. The aforementioned antibody-peptide fusion protein.

36. The antibody-peptide fusion protein according to claim 35, wherein VL comprises the amino acid sequence described in SEQ ID NO: 36, and VH comprises the amino acid sequence described in SEQ ID NO:

55.

37. The antibody-peptide fusion protein according to any one of claims 35 or 36, wherein the amyloid-reactive peptide comprises the amino acid sequence described in SEQ ID NO: 2.

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