US2022411489A1PendingUtilityA1

Modified immunoglobulins for targeting amyloid deposits

69
Assignee: ATTRALUS INCPriority: Nov 15, 2019Filed: Nov 13, 2020Published: Dec 29, 2022
Est. expiryNov 15, 2039(~13.3 yrs left)· nominal 20-yr term from priority
Inventors:Jonathan S. WallJames S. FosterSpencer Guthrie
C07K 2317/92C07K 2317/24A61P 25/28A61K 38/00C07K 2319/30C07K 2317/94C12N 15/62A61K 2039/505G01N 33/6896C07K 2317/52C07K 16/18C07K 2319/33C07K 2317/55C07K 14/4711C07K 2317/622A61K 47/6843C07K 2319/00C07K 2319/01C07K 2317/565G01N 2800/2821C12N 15/85
69
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Claims

Abstract

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.

Claims

exact text as granted — not AI-modified
We claim: 
     
         1 . A modified immunoglobulin, comprising:
 an amyloid reactive peptide; and   an Ig antibody or functional fragment thereof that binds to a human amyloid fibrils, wherein the Ig antibody or functional fragment thereof comprises a heavy chain and a light chain, wherein the peptide and the Ig antibody or functional fragment thereof are joined together at the N-terminal end of the Ig light chain and/or the N- and/or C-terminal end of the Ig heavy chain.   
     
     
         2 . The modified immunoglobulin of  claim 1 , wherein the amyloid reactive peptide comprises an amino acid sequence having at least 85% sequence identity to any one of the amino acid sequences set forth as SEQ ID NOS:1-14. 
     
     
         3 . The modified immunoglobulin of  claim 1  or  claim 2 , wherein the amyloid-reactive peptide and the Ig antibody or functional fragment thereof are joined together at the N-terminal end of the Ig light chain or the N-terminal end of the Ig heavy chain. 
     
     
         4 . The modified immunoglobulin of any one of  claims 1 - 3 , wherein the modified immunoglobulin comprises a spacer sequence between the amyloid-reactive peptide and the Ig antibody or functional fragment thereof. 
     
     
         5 . The modified immunoglobulin any of  claims 1 - 4 , wherein the modified immunoglobulin comprises at least two amyloid-reactive peptides and wherein the amyloid-reactive peptides are the same peptide or different peptides. 
     
     
         6 . The modified immunoglobulin of any one of  claims 1 - 5 ,
 wherein the Ig antibody or functional fragment thereof comprises a light chain variable region (VL) and a heavy chain variable region (VH),   wherein the VL comprises a CDRL1 set forth in SEQ ID NO:20, A CDRL2 set forth in SEQ ID NO: 21, and a CDRL3 set forth in SEQ ID NO: 22; and   wherein the VH comprises a CDRH1 set forth in SEQ ID NO: 17, a CDRH2 set forth in SEQ ID NO: 18, a CDRH3 set forth in SEQ ID NO: 19.   
     
     
         7 . The modified immunoglobulin of  claim 6 , wherein the Ig antibody or functional fragment thereof is a chimeric antibody or functional fragment thereof. 
     
     
         8 . The modified immunoglobulin of any one of  claims 1 - 5 , wherein the Ig antibody or functional fragment thereof comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein
 a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19; or   b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19;   c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         9 . The modified immunoglobulin of any one of  claims 1 - 6  and  8  wherein the Ig antibody or functional fragment thereof comprises human framework sequences. 
     
     
         10 . The modified immunoglobulin of any one of  claims 1 - 9 , wherein the Ig antibody comprises a human Fc region. 
     
     
         11 . The modified immunoglobulin any one of  claims 1 - 6 ,  8  and  10 , wherein the modified immunoglobulin is humanized. 
     
     
         12 . The modified immunoglobulin of any one of  claims 1 - 11 , wherein the modified immunoglobulin binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, A1λ1, or ATTR fibrils. 
     
     
         13 . An antibody-peptide fusion protein comprising an antibody that binds human amyloid fibrils fused to an amyloid-reactive peptide, wherein the antibody comprises a light chain variable region (VL), and a heavy chain variable region (VH). 
     
     
         14 . The antibody-peptide fusion protein of  claim 13 ,
 wherein the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         15 . The antibody-peptide fusion protein of  claim 13  or  14 , wherein the antibody is a chimeric antibody or humanized antibody. 
     
     
         16 . The antibody-peptide fusion protein of  claim 13 , wherein
 a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19; or   b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19;   c) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         17 . The antibody-peptide fusion protein of  claim 14  or  16 ,
 wherein the VL comprises one or more amino acid residues selected from the group consisting of:
 a. Tyr at position 36; 
 b. Leu at position 37; 
 c. Leu at position 46; 
 d. Leu at position 85; and 
 e. Phe at position 87 
 
 wherein the VH comprises one or more amino acid residues selected from the group consisting of:
 a. Val at position 37; 
 b. Leu at position 48; 
 c. Leu at position 67; 
 d. Ser at position 68; 
 e. Lys at position 71; 
 f. Ser at position 76; 
 g. Val at position 78; 
 h. Leu at position 79; 
 i. Phe at position 80; 
 j. Thr at position 89; 
 k. Val at position 93; and 
 l. Thr at position 94 
 
 wherein the amino acid positions are numbered according to the numbering system of Kabat. 
 
     
     
         18 . The antibody-peptide fusion protein of  claim 17 , wherein the VL comprises Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and the VH comprises 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. 
     
     
         19 . The antibody-peptide fusion protein of  claim 17 , wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises 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. 
     
     
         20 . The antibody-peptide fusion protein of  claim 17 , wherein the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:32-42 
     
     
         21 . The antibody-peptide fusion protein of  claim 17  or  20 , wherein the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:43-63. 
     
     
         22 . The antibody-peptide fusion protein of  claim 17 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:34, and the VH comprises an amino acid sequence set forth in SEQ ID NO:48. 
     
     
         23 . The antibody-peptide fusion protein of  claim 17 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:35, and the VH comprises an amino acid sequence set forth in SEQ ID NO:51. 
     
     
         24 . The antibody-peptide fusion protein of any one of  claims 13 - 23 , wherein the amyloid-reactive peptide comprises the amino acid sequence set forth in SEQ ID NO:1-14. 
     
     
         25 . The antibody-peptide fusion protein of any one of  claim 13 - 24 , wherein the amyloid reactive peptide is fused to the N-terminus of the VL or the N-terminus of the VH. 
     
     
         26 . The antibody-peptide fusion protein of  claim 25 , wherein the amyloid reactive peptide is fused to the N-terminus of the VL by a spacer or the N-terminus of the VH. 
     
     
         27 . The antibody-peptide fusion protein of  claim 26 , wherein the spacer is a peptide spacer. 
     
     
         28 . The antibody-peptide fusion protein of  claim 27 , wherein the spacer comprises the amino acid sequence GGGYS. 
     
     
         29 . The antibody-peptide fusion of any one of  claims 13 - 28  wherein the antibody-peptide fusion protein binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, A1λ1, or ATTR fibrils. 
     
     
         30 . A pharmaceutical composition comprising the modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29 . 
     
     
         31 . Nucleic acid(s) encoding the modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29 . 
     
     
         32 . A vector comprising the nucleic acid(s) of  claim 31 . 
     
     
         33 . A host cell comprising the vector of  claim 32 . 
     
     
         34 . A method of making a modified immunoglobulin or an antibody-peptide fusion protein comprising culturing the host cell of  claim 33  under conditions suitable for expression of the vector encoding the modified immunoglobulin or antibody-peptide fusion protein and recovering the modified immunoglobulin or antibody-peptide fusion protein. 
     
     
         35 . A method of treating a subject having an amyloid related disorder, comprising administering to the subject an effective amount of the modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion of any one of  claims 13 - 29 . 
     
     
         36 . The method of  claim 35 , wherein the amyloid related disorder is amyloidosis. 
     
     
         37 . The method of  claim 35 , 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, ACal, AMed, AIAPP, APro, AIns, APrP, or Aβ amyloidosis. 
     
     
         38 . The method of any one of  claims 35 - 37 , wherein the subject is a human. 
     
     
         39 . A method of targeting an amyloid deposit for clearance, comprising contacting an amyloid deposit with the modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29 . 
     
     
         40 . The method of  claim 39 , wherein the amyloid deposit is removed. 
     
     
         41 . The method of  claim 39  or  40 , wherein the amyloid deposit is opsonized by the modified immunoglobulin or the antibody-peptide fusion protein. 
     
     
         42 . A method of targeting an amyloid deposit for clearance, comprising contacting an amyloid deposit with the modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29 . 
     
     
         43 . The method of  claim 42 , wherein targeting the amyloid deposit for clearance results in clearance of the amyloid deposit. 
     
     
         44 . The method of  claim 42  or  43 , wherein clearance results from opsonization of the amyloid deposit. 
     
     
         45 . The method of any of  claims 42 - 44 , wherein the half-life of the amyloid reactive peptide in the modified immunoglobulin or antibody-peptide fusion protein is increased by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or more as compared to the amyloid-reactive peptide alone. 
     
     
         46 . A method for generating a modified immunoglobulin, 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 functional fragment thereof; 
 wherein the second expression vector comprises a second nucleic acid sequence encoding an Ig antibody heavy chain or functional fragment thereof; and 
 wherein the first expression vector and/or the second expression vector comprise a third nucleic acid sequence that encodes 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 and second expression vectors into a cell, wherein expression of the first and second expression vectors in the cell results in an immunoglobulin that is joined to the first peptide.   
     
     
         47 . The method of  claim 46 , wherein the first expression vector and/or the second expression vector comprise a fourth nucleic acid sequence that encodes a second peptide, the fourth nucleic acid sequence being located adjacent to the first nucleic acid sequence and/or the second nucleic acid sequence. 
     
     
         48 . The method of  claim 46 , wherein expression of the first and second expression vectors in the cell results in an immunoglobulin that is joined to the first peptide and the second peptide. 
     
     
         49 . The method of  claim 46 , wherein a 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. 
     
     
         50 . A method of treating a subject suffering from, or suspected to be suffering from, an amyloid-based disease, comprising:
 a) determining whether the subject has an amyloid deposit by:
 i) administering the modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29  to the subject, wherein the modified immunoglobulin or antibody-peptide fusion protein comprises a detectable label and 
 ii) determining whether a signal associated with the detectable label can be detected from the subject; and 
   b) if the signal is detected, administering to the subject an amyloidosis treatment.   
     
     
         51 . The method of  claim 50 , wherein, if a signal is not detected, monitoring the subject for a later development of an amyloid deposit. 
     
     
         52 . The method of  claim 51 , further comprising determining the intensity of the signal and comparing the signal to a threshold value, above which the subject is determined to possess an amyloid deposit. 
     
     
         53 . The method of any of  claims 50 - 52 , wherein the amyloidosis treatment comprises administering the modified immunoglobulin of any of  claims 1 - 12  or the antibody-peptide fusion protein of  claims 13 - 29  to the subject. 
     
     
         54 . The method of  claim 53 , wherein administration of the modified immunoglobulin or the antibody-peptide fusion protein results in clearance of the amyloid deposit in the subject. 
     
     
         55 . A method of identifying an amyloid deposit in a subject, comprising administering the modified immunoglobulin of any one of  claims 1 - 12  or antibody-peptide fusion protein of any one of  claims 13 - 29  to the subject, wherein the modified immunoglobulin or antibody-peptide fusion protein comprises a detectable label and detecting a signal from the modified immunoglobulin or antibody peptide fusion protein. 
     
     
         56 . The method of any of  claims 50 - 55 , wherein the subject is determined to be amyloid free or suffering from monoclonal gammopathy of unknown significance (MGUS), multiple myeloma (MM), or one or more related plasma cell diseases. 
     
     
         57 . A method of detecting a ligand, comprising:
 contacting the ligand with the modified immunoglobulin of any one of  claims 1 - 12  or antibody-peptide fusion protein of any one of  claims 1 - 29 , wherein the modified immunoglobulin or antibody-peptide fusion protein comprises a detectable label, wherein the peptide of the modified immunoglobulin or antibody-peptide fusion protein has binding affinity to the ligand and,   determining a signal from the detectable label, thereby detecting the ligand.   
     
     
         58 . A humanized antibody that binds to human amyloid fibrils, wherein the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein
 the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:20, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and   the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19,   wherein the VL comprises one or more amino acid residues selected from the group consisting of:
 a. Tyr at position 36; 
 b. Leu at position 37; 
 c. Leu at position 46; 
 d. Leu at position 85; and 
 e. Phe at position 87 
   wherein the VH comprises one or more amino acid residues selected from the group consisting of:
 a. Val at position 37; 
 b. Leu at position 48; 
 c. Leu at position 67; 
 d. Ser at position 68; 
 e. Lys at position 71; 
 f. Ser at position 76; 
 g. Val at position 78; 
 h. Leu at position 79; 
 i. Phe at position 80; 
 j. Thr at position 89; 
 k. Val at position 93; and 
 l. Thr at position 94; 
   wherein the amino acid positions are numbered according to the numbering system of Kabat.   
     
     
         59 . The humanized antibody of  claim 58 , wherein the VL comprises Tyr at position 36, Leu at position 37, Leu at position 46, Leu at position 85, and Phe at position 87, and the VH comprises 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. 
     
     
         60 . The humanized antibody of  claim 58 , wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises 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. 
     
     
         61 . The humanized antibody of  claim 58 , wherein the VL comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:33-42. 
     
     
         62 . The humanized antibody of  claim 58  or  61 , wherein the VH comprises an amino acid sequence set forth in the group consisting of SEQ ID NOs:44-63. 
     
     
         63 . The humanized antibody of  claim 58 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:34, and the VH comprises an amino acid sequence set forth in SEQ ID NO:48. 
     
     
         64 . The humanized antibody of  claim 58 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:35, and the VH comprises an amino acid sequence set forth in SEQ ID NO:51. 
     
     
         65 . A humanized antibody that binds to human amyloid fibrils, wherein the humanized antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein
 a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64-70, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:18, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19; or   b) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 20; a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 71-81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         66 . The humanized antibody of  claim 65 , wherein VL comprises one or more amino acid residues selected from the group consisting of:
 a. Tyr at position 36;   b. Leu at position 37;   c. Leu at position 46;   d. Leu at position 85; and   e. Phe at position 87   
       wherein the VH comprises one or more amino acid residues selected from the group consisting of:
 a. Val at position 37; 
 b. Leu at position 48; 
 c. Leu at position 67; 
 d. Ser at position 68; 
 e. Lys at position 71; 
 f. Ser at position 76; 
 g. Val at position 78; 
 h. Leu at position 79; 
 i. Phe at position 80; 
 j. Thr at position 89; 
 k. Val at position 93; and 
 l. Thr at position 94; 
 
       wherein the amino acid positions are numbered according to the numbering system of Kabat. 
     
     
         67 . The humanized antibody of any one of  claims 58 - 66 , wherein the antibody is a full-length antibody, a Fab fragment, or a scFv. 
     
     
         68 . The humanized antibody of any one of  claims 58 - 66 , wherein the antibody comprises an Fc region. 
     
     
         69 . The humanized antibody of  claim 68 , wherein the Fc region is of an IgG1, IgG2, IgG3, or IgG4 isotype. 
     
     
         70 . The humanized antibody of any one of  claims 58 - 69 , wherein the humanized antibody or is conjugated to a detectable label. 
     
     
         71 . The humanized antibody of any one of  claims 58 - 70 , wherein the humanized antibody binds to rVλ6Wil fibrils, Per125 wtATTR extract, KEN hATTR extract, SHI ALλ liver extract, and/or TAL ALκ liver extract. 
     
     
         72 . The humanized antibody of any one of  claims 58 - 71 , wherein the humanized antibody binds to rVλ6Wil, Aβ, Aβ(1-40), IAAP, ALκ4, A1λ1, or ATTR fibrils. 
     
     
         73 . A pharmaceutical composition comprising the humanized antibody of any one of  claims 58 - 72 . 
     
     
         74 . Nucleic acid(s) encoding the humanized antibody of any one of  claims 58 - 73 . 
     
     
         75 . A vector comprising the nucleic acid(s) of  claim 74 . 
     
     
         76 . A host cell comprising the vector of  claim 75 . 
     
     
         77 . A method of making a humanized antibody comprising culturing the host cell of  claim 76  under conditions suitable for expression of the vector encoding the humanized antibody and recovering the humanized antibody. 
     
     
         78 . A method of treating a subject having an amyloid related disorder, comprising administering to the subject an effective amount of the humanized antibody of any one of  claims 58 - 73 . 
     
     
         79 . The method of  claim 78 , wherein the amyloid related disorder is amyloidosis. 
     
     
         80 . The method of  claim 78 , 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, ACal, AMed, AIAPP, APro, AIns, APrP, or Aβ amyloidosis. 
     
     
         81 . The method of any one of  claims 78 - 80 , wherein the subject is a human. 
     
     
         82 . A method of treating a subject suffering from, or suspected to be suffering from, an amyloid-based disease, comprising:
 determining whether the subject has an amyloid deposit by:
 detectably labeling the humanized antibody of any one of  claims 58 - 73 , 
 administering the humanized antibody to the subject, 
 determining whether a signal associated with the detectable label can be detected from the subject; and, 
 if the signal is detected, administering to the subject an amyloidosis treatment. 
   
     
     
         83 . The method of  claim 82 , wherein, if a signal is not detected, monitoring the subject for a later development of an amyloid deposit. 
     
     
         84 . The method of  claim 83 , further comprising determining the intensity of the signal and comparing the signal to a threshold value, above which the subject is determined to possess an amyloid deposit. 
     
     
         85 . The method of any of  claims 82 - 84 , wherein the amyloidosis treatment comprises administering the humanized antibody of any one of  claims 58 - 73  to the subject. 
     
     
         86 . A method of identifying an amyloid deposit in a subject, comprising detectably labeling the humanized antibody of any one of  claims 58 - 73 , administering the humanized antibody to the subject, and detecting a signal from the humanized antibody. 
     
     
         87 . The method of any of  claims 82 - 86 , wherein the subject is determined to be amyloid free or suffering from monoclonal gammopathy of unknown significance (MGUS), multiple myeloma (MM), or one or more related plasma cell diseases. 
     
     
         88 . The modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29 , wherein the modified immunoglobulin or antibody-peptide fusion protein is conjugated to a detectable label. 
     
     
         89 . The modified immunoglobulin of any one of  claims 1 - 12  or the antibody-peptide fusion protein of any one of  claims 13 - 29 , wherein the modified immunoglobulin or antibody-peptide fusion protein comprises a spacer that is N-terminal to the amyloid reactive peptide. 
     
     
         90 . The antibody-peptide fusion protein of  claim 16 , wherein
 a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         91 . The antibody-peptide fusion protein of  claim 90 , wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises 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. 
     
     
         92 . The antibody-peptide fusion protein of  claim 90 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:36, and the VH comprises an amino acid sequence set forth in SEQ ID NO:55. 
     
     
         93 . The antibody-peptide fusion protein of any one of  claims 90 - 92 , wherein the amyloid reactive peptide comprises an amino acid sequence set forth in the group consisting of SEQ ID NO:1-14. 
     
     
         94 . The humanized antibody of  claim 65 , wherein
 a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         95 . The humanized antibody of  claim 94 , wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises 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. 
     
     
         96 . The humanized antibody of any  claim 94 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:36, and the VH comprises an amino acid sequence set forth in SEQ ID NO:55. 
     
     
         97 . The modified immunoglobulin of  claim 8 , wherein
 a) the VL comprises a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:64, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:21, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO:22, and the VH comprises a CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:17, a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:19.   
     
     
         98 . The modified immunoglobulin of  claim 97 , wherein the VL comprises Leu at position 46 and Phe at position 87, and the VH comprises 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. 
     
     
         99 . The modified immunoglobulin of  claim 97 , wherein the VL comprises an amino acid sequence set forth in SEQ ID NO:36, and the VH comprises an amino acid sequence set forth in SEQ ID NO:55. 
     
     
         100 . The modified immunoglobulin of any one of  claims 97 - 99 , wherein the amyloid reactive peptide comprises an amino acid sequence set forth in the group consisting of SEQ ID NO:1-14.

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