Monoclonal antibody targeting lambda subimmunoglobulin light chain conformation specific epitope

Abstract

Provided herein are anti-human lambda light chain antibodies or human lambda light chain binding antibody fragments that bind to human lambda light chain specific epitopes. Also provided are polynucleotides and vectors encoding the same, and compositions comprising anti-human lambda light chain antibodies or human lambda light chain binding antibody fragments. An anti-human lambda light chain antibody or a human lambda light chain binding antibody fragment can be used to measure a human lambda light chain in a biological sample, including contacting the sample with an anti-human lambda light chain antibody or a human lambda light chain binding antibody fragment. Anti-human lambda light chain antibodies or human lambda light chain binding antibody fragments may also be used to reduce lambda light chain aggregates.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 351,533, filed on June 13, 2022, which is incorporated herein by reference in its entirety. Background Art

[0003] Light chain amyloidosis (AL amyloidosis) is a life-threatening secondary condition associated with the primary lineage disorder plasma cell dyscrasia. It is a widely underdiagnosed disease caused by the misfolding of free immunoglobulin light chains (LC) that can be deposited in the heart, kidneys, and nervous system as light chain amyloid (AL). AL amyloidosis occurs in ~15% of plasma cell dyscrasias, in which a single plasma cell clone undergoes unregulated growth, resulting in abnormal production and secretion of large amounts of excess monoclonal free immunoglobulin kappa (κ) or lambda (λ) light chain proteins into the circulation. This increase in light chain protein concentration can lead to the aggregation and accumulation of a specific type of amyloid deposition. Deposition and infiltration of amyloid in vital organs such as the heart, kidneys, and nervous system leads to restrictive cardiomyopathy, nephrotic syndrome, hepatomegaly, peripheral neuropathy, and ultimately death.

[0004] AL amyloidosis is the most common type of systemic amyloidosis. It occurs in 9-14 people per million per year in the Western world. Its prevalence may be underestimated because most patients are not diagnosed until after autopsy. The Amyloidosis Foundation estimates that there are more than 35,000-45,000 patients in the United States and the European Union with this disease, and approximately ~12,000 and ~18,600 new AL patients are diagnosed each year in the United States and the European Union, respectively. The life expectancy of patients diagnosed with AL amyloidosis depends on the extent of organ involvement. For patients with cardiac amyloid deposits (approximately 50%), the prognosis is poor, with a median survival of <6 months, if the patient remains untreated from the onset of heart failure symptoms. Summary of the invention

[0005] Epitopes of human lambda (λ) light chains and antibodies that bind to human λ epitopes are described herein. The epitopes described herein are advantageous because they allow the generation and / or screening of antibodies that bind to and promote the removal of misfolded λ light chains (which are pathogens of λ light chain amyloidosis). One obstacle to the treatment of light chain amyloidosis is that different patients produce light chains with different amino acid sequences, and the binding of anti-light chain antibodies currently targeting variable regions to these amino acid sequences is not equally good. The antibodies described herein target epitopes in the constant region of λ light chains, and in some cases, allow more consistent clinical results and less variability in outcomes between patients. The universality of binding makes the use of anti-human λ light chain antibodies described herein as part of a diagnostic test for light chain amyloidosis have additional benefits. In addition, the anti-human λ light chain antibodies described herein are specific to misfolded light chains, and are almost non-reactive to correctly folded light chains, thereby reducing the chance of unwanted immune side effects due to reduced antibody levels, and avoiding the isolation of potential therapeutic agents caused by unwanted off-target binding that would hinder efficacy. Provided herein are anti-λ light chain antibodies and compositions comprising the anti-λ light chain antibodies for use in treating or diagnosing λ light chain amyloidosis.

[0006] Provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: (a) a heavy chain complementary determining region 1 (H-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:3; (b) a heavy chain complementary determining region 2 (H-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:4; (c) a heavy chain complementary determining region 3 (H-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:5; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:6; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:7; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:8. In some embodiments, the anti-human λ light chain antibody or human λ light chain binding fragment comprises: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or a light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3. In some embodiments:

[0007] In certain aspects, the heavy chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:1; and / or the light chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:2.

[0008] Also provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: (a) a heavy chain complementary determining region 1 (H-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:13; (b) a heavy chain complementary determining region 2 (H-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:14; (c) a heavy chain complementary determining region 3 (H-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:15; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:16; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:17; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:18. In some embodiments, the anti-human λ light chain antibody or human λ light chain binding fragment thereof comprises: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or a light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 11; and / or the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 12.

[0009] Further provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: (a) a heavy chain complementary determining region 1 (H-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:23; (b) a heavy chain complementary determining region 2 (H-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:24; (c) a heavy chain complementary determining region 3 (H-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:25; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:26; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:27; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:28.

[0010] In some embodiments, the anti-human λ light chain antibody or human λ light chain binding fragment thereof comprises: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or a light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 21; and / or the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 22.

[0011] Provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: (a) a heavy chain complementary determining region 1 (H-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:33; (b) a heavy chain complementary determining region 2 (H-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:34; (c) a heavy chain complementary determining region 3 (H-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:35; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:36; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:37; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:38. In some embodiments, the anti-human λ light chain antibody or human λ light chain binding fragment thereof comprises: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or a light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 31; and / or the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 32.

[0012] Also provided and described herein are anti-human λ light chain antibodies or human λ light chain binding antibody fragments thereof, which comprise: (a) a heavy chain complementary determining region 1 (H-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:43; (b) a heavy chain complementary determining region 2 (H-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:44; (c) a heavy chain complementary determining region 3 (H-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:45; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence set forth in SEQ ID NO:46; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence set forth in SEQ ID NO:47; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence set forth in SEQ ID NO:48. In some embodiments, the anti-human λ light chain antibody or human λ light chain binding fragment comprises: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or a light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 41; and / or the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 42.

[0013] Provided and described herein are anti-human λ light chain antibodies or human λ light chain antibody fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 1; and / or a light chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 2. Also provided are anti-human λ light chain antibodies or human λ light chain binding antibody fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 1; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 2.

[0014] Provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 11; and / or a light chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 12. Also provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 11; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 12.

[0015] Provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 21; and / or a light chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 22. Also provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 21; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 22.

[0016] Provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 31; and / or a light chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 32. Also provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 31; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 32.

[0017] Provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 41; and / or a light chain variable region comprising an amino acid sequence having at least about 90%, 95%, 97%, 98% or 99% identity to SEQ ID NO: 42. Also provided and described herein are anti-human λ light chain antibodies or human λ light chain binding fragments thereof, comprising: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 41; and / or a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 42.

[0018] In some embodiments, the anti-human λ light chain antibody is a monoclonal antibody. In some embodiments, the anti-human λ light chain antibody is a chimeric antibody, a humanized antibody or a human antibody. In some embodiments, the human λ light chain binding antibody fragment includes Fab, Fab', F(ab')2, scFv, dsFv, ds-scFv, dimer, mini antibody or bispecific antibody (diabody). In some embodiments, the anti-human λ light chain antibody or its human λ light chain binding fragment is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 isotype. In some embodiments, the IgG antibody is an IgG4 isotype. In some embodiments, the anti-human λ light chain antibody or its human λ light chain binding fragment binds to an epitope comprising SEQ ID NO: 51. In some embodiments, the binding to the epitope comprising SEQ ID NO: 51 is measured by ELISA, surface plasmon resonance, biolayer interferometry or isothermal calorimetry.

[0019] In some embodiments, the anti-human λ light chain antibody or its human λ light chain binding fragment is coupled to a tag. In some embodiments, the tag comprises a chemical tag, a biological tag, a fluorescent tag, a radioisotope tag or any combination thereof.

[0020] Provided herein and described are pharmaceutical compositions comprising the anti-human λ light chain antibody or its human λ light chain binding fragment and a pharmaceutically acceptable excipient, diluent or carrier. Also provided are nucleic acids encoding the anti-human λ light chain antibody or its human λ light chain binding fragment. Further provided are host cells comprising nucleic acids encoding the anti-human λ light chain antibody or human λ light chain binding fragment.

[0021] Provided herein and described are methods for diagnosing amyloidosis in a sample, the method comprising contacting a biological sample obtained from a subject with an anti-human λ light chain antibody or a human λ light chain binding fragment thereof as described herein. Also provided are methods for detecting free light chain polypeptides in a sample, the method comprising contacting the biological sample with an anti-human λ light chain antibody or a human λ light chain binding fragment thereof as described herein. Further provided herein are methods for detecting amyloid deposits in a sample, the method comprising contacting the biological sample with an anti-human λ light chain antibody or a human λ light chain binding fragment thereof as described herein.

[0022] In some embodiments, the sample is from blood, serum, plasma, solid tissue, and any combination thereof. In some embodiments, the sample is from a subject.

[0023] Provided herein and described are methods for promoting the clearance of λ light chain aggregates by monocytes, the methods comprising contacting the λ light chain aggregates with an anti-human λ light chain antibody or its human λ light chain binding fragment described herein. Also provided herein and described are methods for increasing the clearance of λ light chain aggregates, the methods comprising contacting the λ light chain aggregates with an anti-human λ light chain antibody or its human λ light chain binding fragment described herein.

[0024] Provided are methods of reducing lambda light chain aggregates in a subject, the methods comprising administering to the subject an anti-human lambda light chain antibody or human lambda light chain binding fragment thereof described herein, thereby reducing the number of lambda light chain aggregates in the subject.

[0025] Also provided and described herein are methods of treating a disease or condition characterized by lambda light chain aggregates, the methods comprising administering to the subject an anti-human lambda light chain antibody or human lambda light chain binding fragment thereof described herein, thereby reducing the number of lambda light chain aggregates in the subject.

[0026] In some embodiments, the subject suffers from a plasma cell disorder. In some embodiments, the plasma cell disorder comprises plasma cell dyscrasia. In some embodiments, the plasma cell dyscrasia is selected from monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), plasma cell leukemia (PCL), and any combination thereof. In some embodiments, the subject suffers from systemic amyloidosis. In some embodiments, the subject suffers from AL amyloidosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The novel features of the present composition disclosure are particularly set forth in the appended claims. A better understanding of the features and advantages of the present composition disclosure will be obtained by reference to the following detailed description of illustrative embodiments in which the principles of the invention are utilized and the accompanying drawings, in which: Figure 1Shown is the unfolded full-length lambda light chain protein detected by denaturing protein blotting (western blot) after denaturing SDS-PAGE, using five mouse monoclonal antibodies described herein to probe. The sample loaded includes kappa (κ) and lambda (λ) light chains pooled from human myeloma serum, and lambda light chain amyloidosis patient serum (pt). Two of the five clones demonstrated reactivity to both lambda and kappa serum.

[0028] Figure 2A , Figure 2B , Figure 2C , Figure 2D , Figure 2E and Figure 2F is a line graph illustrating the use of the five mouse monoclonal antibodies described in this article ( Figure 2A-2E ) or a commercial control antibody ("Com", Figure 2F ) to detect native or misfolded forms of lambda light chain (Wil). Different concentrations of native or misfolded Wil were bound and detected using appropriate HRP-conjugated secondary antibodies using TMB chemistry.

[0029] Figure 3 Native Western blot images of misfolded, native or amyloid species probed with 5 mouse monoclonal antibodies are shown. Western blots were prepared by transfer from native PAGE and probed with appropriate secondary antibodies. Monomeric species of Wil, either native (n) or misfolded (m), appear as ~25 kDa bands, while aggregates (a) or large MW species (>250 kDa) appear as smears in the amyloid (a) sample lanes.

[0030] Figure 4A and Figure 4B Is a graphical representation of the capture ELISA results using misfolded (half-filled circles) or native (full circles) Wil as the target for monoclonal antibody binding. For these two clones, increasing concentrations of native Wil did not result in significant signal changes, while the same increase in concentration of misfolded Wil resulted in increased binding. These binding curves correspond to affinities of 69 nM (clone 1) and 32 nM (clone 2).

[0031] Figure 5A and Figure 5B is clone 1 ( Figure 5A ) and Clone 2 ( Figure 5B) and the misfolded lambda light chain Wil. The binding of clone 1 and clone 2 to their misfolded Wil was measured by the change in wavelength (nm) over 500 seconds (120 s association, 300 s dissociation). The association and dissociation curves were fitted using Octet Systems software to determine K D .

[0032] Figure 6A-6F Clone 1 (muLX-97) and Wil ( Fig. 6A , Fig.6D )、H3( Figure 6B , Fig. 6E ) and FOR102( Figure 6C , Fig. 6F )Misfolding of lambda light chain variants ( Fig. 6A , Figure 6B , Figure 6C ) and natural ( Fig.6D , Fig. 6E , Fig. 6F ) conformations. The binding of muLX-97 to its misfolded Wil at a range of target concentrations was measured by changes in wavelength (nm) over 720 seconds (120 s association, 600 s dissociation). The association and dissociation curves were fitted using Octet Systems software to determine K D .

[0033] Fig. 7A and Figure 7B is a line graph illustrating misfolding as measured by MST (microscale thermophoresis) Fig. 7A ) or amyloid ( Figure 7B ) Wil binding to muLX-97. The shift in normalized relative fluorescence (Fnorm) during thermophoresis as a function of ligand concentration shows a shift corresponding to K D 61.9 nM for misfolded Wil and 87 nM for amyloid Wil.

[0034] Figure 8A-8D Figure 1 shows an immunohistochemical staining and fluorescence image of a biopsy-proven case of λAL amyloidosis. Fig. 8A , Figure 8B ) for the gastrointestinal tract ( Fig. 8A , Figure 8C ) and soft tissue cases ( Figure 8B , Fig.8D ) were stained (dark areas). The same tissues were co-stained with Congo red amyloid-specific dye ( Figure 8C , Fig.8D) and visualize the fluorescent signal (white area). Scale bar = 100 μm.

[0035] Figure 9A-9G Images of immunohistochemical staining of biopsy-proven λ AL amyloidosis cases or pathological controls using muLX-97 staining. Fig. 9A , Fig. 9B ),kidney( Fig. 9C ) and soft tissue ( Fig.9D , Fig.9E ) cases stained positive (dark areas), but the pathological controls of pancreatic tissues with IAPP amyloid protein ( Fig.9F ) and ATTR cardiac amyloid tissue ( Figure 9G ) staining negative. Scale bar = 100 μm.

[0036] Fig. 10A and 10B The fluorescence images of RAW 264.7 cells in representative fields. Fig. 10A ) or muLX-97( Fig. 10B ) were incubated with pHrodo-tagged Wil for 30 min in the presence of . pHrodo fluorescent signal (white) represents cells that internalized Wil amyloid via phagocytosis. Scale bar = 100 μm.

[0037] Fig.11A and Fig. 11B The fluorescence images of J774.1 cells in representative fields. Fig.11A ) or muLX-97( Fig. 11B ) were incubated with pHrodo-tagged Wil for 30 min in the presence of . pHrodo fluorescent signal (white) represents cells that internalized Wil amyloid via phagocytosis. Scale bar = 100 μm.

[0038] Fig. 12A and Fig. 12B is a line graph showing the dose-dependent phagocytic uptake of pHrodo-tagged amyloid Wil by muLX-97 in mouse macrophages. 40,000 RAW 264.7 ( Fig. 12A ) or J774.1( Fig. 12B) Cells were incubated with pHrodo-tagged Wil and isotype antibodies (circles) or muLX-97 (triangles) at increasing antibody concentrations. Cells with pHrodo fluorescence were imaged (white) and quantified using a Biotek Cytation 5 imager. Total cell counts were performed using the nuclear stain Nuc Blue live cell reagent to determine pHrodo positive %. The pHrodo positive cell % was plotted as a function of muLX-97 concentration, showing the percentage of cells corresponding to EC 50 A shift of 20 nM for RAW 264.7 cells and 7 nM for J774.1 cells. DETAILED DESCRIPTION

[0039] Current treatments for AL amyloidosis are therapies designed to treat underlying plasma cell dyscrasias (i.e., multiple myeloma) by targeting cells that produce toxic light chains. The only currently available on-label therapy is the monoclonal antibody-based therapy daratumumab and hyaluronidase-fihj combined with bortezomib, cyclophosphamide, and dexamethasone. The antibody (and other off-label therapies) target the CD38 receptor on plasma cells to induce antibody-dependent cellular toxicity. The drug's initial indication was multiple myeloma, but it was approved in January 2021 as an on-label treatment for newly diagnosed AL amyloidosis patients. Other treatments for AL amyloidosis include more invasive interventions such as autologous stem cell transplantation (ASTC) or standard cytotoxic chemotherapy. However, all previous therapies work by targeting the proliferation of pathogenic plasma cell clones and do not address pre-existing amyloid deposits that directly affect organs. Although effective for early patients, daratumumab is also not recommended for patients with advanced heart disease. Therefore, non-invasive therapies that do not deplete plasma cell populations to resolve existing toxic amyloid deposits to improve organ function in advanced patients remain a major challenge and unmet need for the treatment of AL amyloidosis and / or conditions characterized by misfolded antibody molecules.

[0040] Anti-human lambda light chain antibody

[0041] Provided herein are antibodies and antibody fragments that bind to human lambda light chains. Anti-human lambda light chain antibodies described herein are useful and advantageous because antibodies and antibody fragments specifically bind to misfolded lambda light chains (i.e., as compared to folded lambda light chains). In general, their usefulness and advantages can be attributed to the ability of anti-human lambda light chain antibodies to specifically bind to the specific epitope SEQ ID NO of misfolded lambda light chains: 51. The antibodies provided herein are described as anti-human lambda light chain antibodies, which in some cases promote the removal of misfolded lambda light chains (e.g., present in AL amyloidosis or plasma cell disorders). In some embodiments, anti-human lambda light chain antibodies can be used to reduce amyloid deposits and / or treat a method for a disease characterized by amyloid deposits (e.g., present in AL amyloidosis or plasma cell disorders). In other cases, anti-human lambda light chain antibodies can be used for diagnosis, detection or screening of misfolded lambda light chains, amyloid deposits and / or disorders associated therewith.

[0042] Anti-human lambda light chain CDR

[0043] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0044] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 3;

[0045] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 4;

[0046] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 5;

[0047] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO:6;

[0048] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 7; and / or

[0049] (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:8.

[0050] In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 1; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 2. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0051] Also provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0052] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 13;

[0053] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 14;

[0054] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 15;

[0055] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 16;

[0056] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 17; and / or

[0057] (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:18.

[0058] In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 11; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 12. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0059] Further provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0060] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 23;

[0061] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 24;

[0062] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 25;

[0063] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 26;

[0064] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 27; and / or

[0065] (f) a light chain complementarity determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 28.

[0066] In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 21; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 22. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0067] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0068] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 33;

[0069] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 34;

[0070] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 35;

[0071] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 36;

[0072] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 37; and / or

[0073] (f) a light chain complementarity determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 38.

[0074] In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 31; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 32. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0075] Further provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0076] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 43;

[0077] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO:44;

[0078] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO:45;

[0079] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO:46;

[0080] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 47; and / or

[0081] (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:48.

[0082] In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 41; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 42. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0083] Further provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0084] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 33 or 43;

[0085] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 34 or 44;

[0086] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 35 or 45;

[0087] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 36 or 46;

[0088] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 37 or 47; and / or

[0089] (f) a light chain complementarity determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 38 or 48;

[0090] wherein the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 31 or 41; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 32 or 42. In some embodiments, the anti-human λ light chain antibody and / or human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0091] Also provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0092] (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence NFGMH;

[0093] (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence WMNT YTGESTYAD(D or E)FKG;

[0094] (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence ISYDYD GYTMD (F or Y);

[0095] (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence RS(G or S)QSLVH(R or S)NGNTY(F or L)H;

[0096] (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence K(V or L)SI(R or N)FS; and / or

[0097] (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence SQSTHV PWT, wherein the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51. In some embodiments, the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment comprises: a heavy chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 31 or 41; and / or a light chain variable region comprising an amino acid sequence having at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 98% identity to SEQ ID NO: 32 or 42. In some embodiments, the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51.

[0098] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0099] (a) heavy chain complementarity determining region 1 (H-CDR1);

[0100] (b) heavy chain complementarity determining region 2 (H-CDR2);

[0101] (c) heavy chain complementarity determining region 3 (H-CDR3);

[0102] (d) light chain complementarity determining region 1 (L-CDR1);

[0103] (e) light chain complementarity determining region 2 (L-CDR2); and / or

[0104] (f) light chain complementarity determining region 3 (L-CDR3);

[0105] Wherein H-CDR1, H-CDR2 and H-CDR3 are derived from SEQ ID NO:1, and L-CDR1, L-CDR2 and L-CDR3 are derived from SEQ ID NO:2; and wherein H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia, Kabat, IMGT, Contact or AbM methods. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Kabat. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using IMGT. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using Contact. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using AbM.

[0106] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0107] (a) heavy chain complementarity determining region 1 (H-CDR1);

[0108] (b) heavy chain complementarity determining region 2 (H-CDR2);

[0109] (c) heavy chain complementarity determining region 3 (H-CDR3);

[0110] (d) light chain complementarity determining region 1 (L-CDR1);

[0111] (e) light chain complementarity determining region 2 (L-CDR2); and / or

[0112] (f) light chain complementarity determining region 3 (L-CDR3);

[0113] Wherein H-CDR1, H-CDR2 and H-CDR3 are derived from SEQ ID NO: 11, and L-CDR1, L-CDR2 and L-CDR3 are derived from SEQ ID NO: 12; and wherein H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia, Kabat, IMGT, Contact or AbM methods. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Kabat. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using IMGT. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using Contact. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using AbM.

[0114] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0115] (a) heavy chain complementarity determining region 1 (H-CDR1);

[0116] (b) heavy chain complementarity determining region 2 (H-CDR2);

[0117] (c) heavy chain complementarity determining region 3 (H-CDR3);

[0118] (d) light chain complementarity determining region 1 (L-CDR1);

[0119] (e) light chain complementarity determining region 2 (L-CDR2); and / or

[0120] (f) light chain complementarity determining region 3 (L-CDR3);

[0121] Wherein H-CDR1, H-CDR2 and H-CDR3 are derived from SEQ ID NO:21, and L-CDR1, L-CDR2 and L-CDR3 are derived from SEQ ID NO:22; and wherein H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia, Kabat, IMGT, Contact or AbM methods. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Kabat. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using IMGT. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using Contact. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using AbM.

[0122] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0123] (a) heavy chain complementarity determining region 1 (H-CDR1);

[0124] (b) heavy chain complementarity determining region 2 (H-CDR2);

[0125] (c) heavy chain complementarity determining region 3 (H-CDR3);

[0126] (d) light chain complementarity determining region 1 (L-CDR1);

[0127] (e) light chain complementarity determining region 2 (L-CDR2); and / or

[0128] (f) light chain complementarity determining region 3 (L-CDR3);

[0129] Wherein H-CDR1, H-CDR2 and H-CDR3 are derived from SEQ ID NO:31, and L-CDR1, L-CDR2 and L-CDR3 are derived from SEQ ID NO:32; and wherein H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia, Kabat, IMGT, Contact or AbM methods. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Kabat. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using IMGT. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using Contact. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using AbM.

[0130] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0131] (a) heavy chain complementarity determining region 1 (H-CDR1);

[0132] (b) heavy chain complementarity determining region 2 (H-CDR2);

[0133] (c) heavy chain complementarity determining region 3 (H-CDR3);

[0134] (d) light chain complementarity determining region 1 (L-CDR1);

[0135] (e) light chain complementarity determining region 2 (L-CDR2); and / or

[0136] (f) light chain complementarity determining region 3 (L-CDR3);

[0137] Wherein H-CDR1, H-CDR2 and H-CDR3 are derived from SEQ ID NO:41, and L-CDR1, L-CDR2 and L-CDR3 are derived from SEQ ID NO:42; and wherein H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia, Kabat, IMGT, Contact or AbM methods. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Chothia. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 are defined using Kabat. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using IMGT. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using Contact. In some embodiments, H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2, and L-CDR3 are defined using AbM.

[0138] In some embodiments, the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 1. In certain embodiments, the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope consisting of SEQ ID NO: 1. In certain embodiments, binding is measured by ELISA, surface plasmon resonance, biolayer interferometry, or isothermal calorimetry.

[0139] In some embodiments, the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment further comprises a heavy chain framework region 1 (H-FR1), a heavy chain framework region 2 (H-FR2), a heavy chain framework region 3 (H-FR3), and a heavy chain framework region 4 (H-FR4); and a light chain framework region 1 (L-FR1), a light chain framework region 2 (L-FR2), a light chain framework region 3 (L-FR3), and a light chain framework region 4 (L-FR4). In some embodiments, the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment further comprises a heavy chain variable region and a light chain variable region. In some embodiments, the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment further comprises a heavy chain constant region and a light chain constant region.

[0140] Anti-human lambda light chain variable region

[0141] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0142] A heavy chain variable region comprising an amino acid sequence that is at least about 70% identical to SEQ ID NO: 1; and / or

[0143] a light chain variable region comprising an amino acid sequence at least about 70% identical to SEQ ID NO: 2;

[0144] Wherein the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51 (e.g., as measured by ELISA). In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 2. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 1, and the light chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 2.

[0145] Also provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0146] A heavy chain variable region comprising an amino acid sequence that is at least about 70% identical to SEQ ID NO: 11; and / or

[0147] a light chain variable region comprising an amino acid sequence at least about 70% identical to SEQ ID NO: 12;

[0148] Wherein the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51 (e.g., as measured by ELISA). In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 12. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 12. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 12. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 12. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 12. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 12. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 11, and the light chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 12.

[0149] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0150] A heavy chain variable region comprising an amino acid sequence that is at least about 70% identical to SEQ ID NO: 21; and / or

[0151] a light chain variable region comprising an amino acid sequence at least about 70% identical to SEQ ID NO:22;

[0152] Wherein the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51 (e.g., as measured by ELISA). In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 22. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 22. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 22. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 22. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 22. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 22. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 21, and the light chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 22.

[0153] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0154] A heavy chain variable region comprising an amino acid sequence that is at least about 70% identical to SEQ ID NO: 31; and / or

[0155] a light chain variable region comprising an amino acid sequence at least about 70% identical to SEQ ID NO:32;

[0156] Wherein the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51 (e.g., as measured by ELISA). In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 32. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 32. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 32. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 32. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 32. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 32. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 31, and the light chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 32.

[0157] Provided and described herein are anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments comprising:

[0158] A heavy chain variable region comprising an amino acid sequence that is at least about 70% identical to SEQ ID NO:41; and / or

[0159] a light chain variable region comprising an amino acid sequence at least about 70% identical to SEQ ID NO:42;

[0160] Wherein the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment binds to an epitope comprising SEQ ID NO: 51 (e.g., as measured by ELISA). In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence having at least about 80% identity with SEQ ID NO: 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence having at least about 85% identity with SEQ ID NO: 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence having at least about 90% identity with SEQ ID NO: 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence having at least about 95% identity with SEQ ID NO: 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO: 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO: 42. In some embodiments, the heavy chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 41, and the light chain variable region comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO: 42.

[0161] Anti-human lambda light chain constant region

[0162] In some embodiments, the anti-human λ light chain antibody comprises a complete or full-length antibody, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA and IgD. In certain embodiments, the anti-human λ light chain antibody comprises a human IgG constant region. In certain embodiments, the anti-human λ light chain antibody comprises a human IgG1 constant region. In certain embodiments, the anti-human λ light chain antibody comprises a human IgG2 constant region. In certain embodiments, the anti-human λ light chain antibody comprises a human IgG3 constant region. In certain embodiments, the anti-human λ light chain antibody comprises a human IgG4 constant region. In some embodiments, the anti-human λ light chain antibody comprises a heavy chain sequence having at least 85% sequence identity with SEQ ID NO: 52. In some embodiments, the anti-human λ light chain antibody comprises a heavy chain sequence having at least 90% sequence identity with SEQ ID NO: 52. In some embodiments, the anti-human λ light chain antibody comprises a heavy chain sequence having at least 95% sequence identity with SEQ ID NO: 52. In some embodiments, the anti-human λ light chain antibody comprises a heavy chain sequence having at least 98% sequence identity to SEQ ID NO: 52. In some embodiments, the anti-human λ light chain antibody comprises a heavy chain sequence having at least 99% sequence identity to SEQ ID NO: 52. In some embodiments, the anti-human λ light chain antibody comprises a heavy chain sequence comprising SEQ ID NO: 52.

[0163] In certain embodiments, antibodies are described herein that bind to misfolded human lambda light chains. In certain embodiments, the antibodies bind to aggregated human lambda light chains. In certain embodiments, the antibodies bind to amyloid human lambda light chains. In certain embodiments, the antibodies bind to amyloid human lambda light chains with a K of less than about 100 nanomolar. D In certain embodiments, the antibody binds to aggregated human lambda light chains with a K of less than about 90 nanomolar. D In certain embodiments, the antibody binds to aggregated human lambda light chains with a K of less than about 80 nanomolar. D In certain embodiments, the antibody binds to aggregated human lambda light chains with a K of greater than about 1, 10, 20, 30, 40, or 50 nanomolar. D In certain embodiments, the antibody binds to aggregated human lambda light chains with a K of less than about 100 nanomolar. D In certain embodiments, the antibody binds to amyloid human lambda light chain with a K of less than about 90 nanomolar. D In certain embodiments, the antibody binds to amyloid human lambda light chain with a K of less than about 80 nanomolar. D In certain embodiments, the antibody binds to amyloid human lambda light chain with a K of less than about 70 nanomolar. D In certain embodiments, the antibody binds to amyloid human lambda light chain with a K of greater than about 1, 10, 20, 30, 40, or 50 nanomolar.D Amyloid-binding human lambda light chain.

[0164] Composition

[0165] Provided herein are compositions (e.g., pharmaceutical compositions) comprising anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments described herein. Compositions or pharmaceutical preparations or pharmaceutical compositions generally encompass and / or refer to preparations whose form allows the biological activity of the active ingredients contained therein to be effective and does not contain additional components that are unacceptably toxic to the subject to which the preparation will be administered. Pharmaceutical preparations are generally sterile (e.g., sterile or free of all living microorganisms and their spores).

[0166] Compositions comprising anti-human lambda light chain antibodies and / or human lambda light chain binding antibody fragments described herein may be included in pharmaceutical compositions comprising one or more pharmaceutically acceptable excipients, carriers, and diluents. In certain embodiments, the antibodies of the present disclosure are suspended in sterile and / or isotonic solutions for administration. In certain embodiments, the solution comprises about 0.9% NaCl. In certain embodiments, the solution comprises about 5.0% dextrose. In certain embodiments, the solution further comprises one or more of: a buffer such as acetate, citrate, histidine, succinate, phosphate, bicarbonate, and hydroxymethylaminomethane (Tris); a surfactant such as polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), and poloxamer 188; a polyol / disaccharide / polysaccharide such as glucose, dextrose, mannose, mannitol, sorbitol, sucrose, L-histidine / L-histidine monohydrochloride monohydrate, trehalose, and dextran 40; an amino acid such as glycine or arginine; an antioxidant such as ascorbic acid, methionine; or a chelating agent such as EDTA or EGTA.

[0167] The formulation for administering the antibody may include one or more of the following: a buffer such as acetate, citrate, histidine, succinate, phosphate, bicarbonate, and hydroxymethylaminomethane (Tris); a surfactant such as polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), and poloxamer 188; a polyol / disaccharide / polysaccharide such as glucose, dextrose, mannose, mannitol, sorbitol, sucrose, trehalose, and dextran 40; an amino acid such as glycine or arginine; an antioxidant such as ascorbic acid, methionine; or a chelating agent such as EDTA or EGTA. In addition, a compound or molecule that reduces pain at the injection site, such as hyaluronidase, may be included, for example, at a concentration of about 2,000 U / ml to about 12,000 U / ml.

[0168] In certain embodiments, the anti-human lambda light chain antibody or human lambda light chain binding antibody fragment described herein is transported / stored by lyophilization and reconstituted before administration. In certain embodiments, the lyophilized antibody formulation comprises a bulking agent such as mannitol, sorbitol, sucrose, trehalose, dextran 40 or a combination thereof. The lyophilized formulation may be contained in a vial consisting of glass or other suitable non-reactive materials. When formulated, the antibody, whether recombinant or not, can be buffered with a certain pH, generally less than 7.0. In certain embodiments, the pH may be between 4.5 and 6.5, 4.5 and 6.0, 4.5 and 5.5, 4.5 and 5.0 or 5.0 and 6.0.

[0169] In some embodiments, the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment is configured for intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, or intrasternal injection. In some embodiments, the composition comprising the anti-human lambda light chain antibody and / or human lambda light chain binding antibody fragment is configured for intravenous injection.

[0170] Also described herein are kits comprising, in a suitable container, one or more antibodies described herein and one or more additional components selected from the group consisting of: instructions for use; diluents, excipients, carriers, and administration devices.

[0171] In certain embodiments, described herein are methods of preparing a treatment comprising mixing one or more pharmaceutically acceptable excipients, carriers or diluents with an anti-human lambda light chain antibody and / or a human lambda light chain binding antibody fragment. In certain embodiments, described herein are methods of preparing a cancer treatment for storage or transport comprising lyophilizing one or more antibodies of the present disclosure.

[0172] Host cells and nucleic acids

[0173] Host cells, host cell lines and / or host cell cultures are used interchangeably and generally include and / or refer to cells into which exogenous nucleic acids have been introduced, including the progeny of such cells. Host cells include transformants and transformed cells, including primary transformed cells and progeny derived therefrom, without regard to the number of passages.

[0174] An isolated nucleic acid generally encompasses and / or refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid encompasses a nucleic acid molecule contained in a cell that normally contains a nucleic acid molecule, but the nucleic acid molecule is present outside the chromosome or in a chromosomal position different from its natural chromosomal position. An isolated nucleic acid encoding an antibody encompasses and / or refers to one or more nucleic acid molecules encoding antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules in a single vector or a separate vector, and such nucleic acid molecules present in one or more positions in a host cell. In some embodiments, provided herein are nucleic acid sequences encoding anti-human λ light chain antibodies and / or human λ light chain binding antibody fragments described herein. In some embodiments, a host cell is also provided, which comprises a nucleic acid sequence encoding an anti-human λ light chain antibody and / or a human λ light chain binding antibody fragment. In some embodiments, the nucleic acid sequence encoding an anti-human λ light chain antibody and / or a human λ light chain binding antibody fragment has at least 60% sequence identity with any one of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, or 49-50. In some embodiments, the nucleic acid sequence encoding the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment has at least 70% sequence identity with any one of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, or 49-50. In some embodiments, the nucleic acid sequence encoding the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment has at least 80% sequence identity with any one of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, or 49-50. In some embodiments, the nucleic acid sequence encoding the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment has at least 90% sequence identity with any one of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, or 49-50. In some embodiments, the nucleic acid sequence encoding the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment has at least 95% sequence identity with any one of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, or 49-50. In some embodiments, the nucleic acid sequence encoding the anti-human λ light chain antibody and / or the human λ light chain binding antibody fragment comprises the sequence of any one of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, or 49-50.

[0175] method

[0176] The anti-human λ light chain antibodies and / or human λ light chain binding antibody fragments described herein can be used to identify, detect and treat misfolded λ light chains (eg, λ light chain aggregates or amyloid deposits) and diseases or disorders associated therewith.

[0177] Methods for detecting light chains

[0178] Provided herein and described is a method for diagnosing amyloidosis in a sample, the method comprising contacting a biological sample obtained from a subject with an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein. Also provided herein is a method for detecting free light chain polypeptides in a sample, the method comprising contacting a biological sample with an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein. Further provided is a method for detecting amyloid deposits in a sample, the method comprising contacting a biological sample with an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein. In some embodiments, the sample is from blood, serum, plasma, solid tissue, and any combination thereof. In some embodiments, the sample is from a subject.

[0179] As described, certain embodiments of the present disclosure are directed to methods for measuring light chain immunoglobulins. Anti-human λ light chain antibodies or human λ light chain binding antibody fragments disclosed herein can be used to measure any immunoglobulin light chain or its aggregate. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to measure immunoglobulin light chains or their aggregates in vivo. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to measure immunoglobulin light chains or their aggregates in biological samples collected from objects. In some embodiments, the biological sample is a plasma sample, a blood sample, a tissue sample (e.g., a tissue biopsy), a urine sample, or any combination thereof.

[0180] In some embodiments, the method includes measuring light chain fibrils. Therefore, the antibodies described herein can be used to identify objects with diseases or conditions characterized by the formation of immunoglobulin light chain deposits (e.g., amyloid fibril deposits). In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to identify (e.g., diagnose) objects suffering from amyloidosis. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to identify objects suffering from systemic amyloidosis. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to identify objects suffering from AL amyloidosis. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to identify objects suffering from plasma cell dyscrasias. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to identify objects suffering from monoclonal gammopathy of unknown significance (MGUS). In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are used to identify objects suffering from multiple myeloma (MM). In some embodiments, an anti-human lambda light chain antibody or a human lambda light chain binding antibody fragment is used to identify a subject as having plasma cell leukemia (PCL).

[0181] In some embodiments, the anti-human λ light chain antibody or human λ light chain binding antibody fragment comprises one or more tags, covalent or non-covalent antibodies or antigen binding fragments. Any tags known in the art can be used to increase the ability of anti-human λ light chain antibodies or human λ light chain binding antibody fragments to be detected and / or visualized. In some embodiments, the label comprises a chemical label. In some embodiments, the label comprises an enzyme label. In some embodiments, the label comprises a substrate detectable in the presence of a specific enzyme. In some embodiments, the label is a fluorescent label. In some embodiments, the label is a radioisotope label.

[0182] Certain embodiments of the present disclosure are directed to methods for measuring light chain immunoglobulins in vivo using anti-human λ light chain antibodies or human λ light chain binding antibody fragments disclosed herein. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments comprise detectable probes. In some embodiments, anti-human λ light chain antibodies or human λ light chain binding antibody fragments are antibodies conjugated with contrast agents. In some embodiments, the method is a diagnostic medical imaging method, such as PET / CT, SPECT, MRI, or any combination thereof.

[0183] Methods for treating and / or reducing misfolded lambda light chains

[0184] Provided herein and described are methods for promoting the removal of λ light chain aggregates by monocytes, the method comprising contacting λ light chain aggregates with anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein. Also provided and described are methods for increasing the removal of λ light chain aggregates, the method comprising contacting λ light chain aggregates with anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein. Further provided and described are methods for reducing λ light chain aggregates in a subject, the method comprising administering to the subject anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein, thereby reducing the number of λ light chain aggregates in the subject. Provided and described are methods for treating diseases or conditions characterized by λ light chain aggregates, the method comprising administering to the subject anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein, thereby reducing the number of λ light chain aggregates in the subject. Provided and described are methods for treating AL amyloidosis (e.g., in subjects with λ light chain aggregates), the method comprising administering to the subject anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein, thereby reducing the number of λ light chain aggregates in the subject.

[0185] In some embodiments, the subject suffers from a plasma cell disorder. In some embodiments, the plasma cell disorder comprises plasma cell dyscrasias. In some embodiments, the plasma cell dyscrasias are selected from monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), plasma cell leukemia (PCL), and any combination thereof. In certain embodiments, the subject is a human.

[0186] Certain uses of anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein are directed to methods for treating a disease or condition in a subject in need thereof, comprising administering an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein. In some embodiments, the disease or condition comprises amyloidosis. In some embodiments, the disease or condition comprises systemic amyloidosis. In some embodiments, the disease or condition comprises AL amyloidosis. In some embodiments, the disease or condition comprises plasma cell dyscrasia. In some embodiments, the disease or condition comprises monoclonal gammopathy of unknown significance (MGUS). In some embodiments, the disease or condition comprises multiple myeloma (MM). In some embodiments, the disease or condition comprises plasma cell leukemia (PCL).

[0187] Certain uses of anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein are directed to reducing and / or removing immunoglobulin light chain aggregates (e.g., amyloid fibrils) in subjects in need, including administering anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein. In some embodiments, the subject has one or more immunoglobulin light chain aggregates, such as amyloid fibrils, in the heart. In some embodiments, the subject has one or more immunoglobulin light chain aggregates, such as amyloid fibrils, in the kidney. In some embodiments, the subject has one or more immunoglobulin light chain aggregates, such as amyloid fibrils, in nervous system tissue. In some embodiments, the subject has one or more immunoglobulin light chain aggregates, such as amyloid fibrils, in the brain.

[0188] Certain uses of the anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein are directed to methods of treating nephrotic syndrome in a subject in need thereof, comprising administering to the subject an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein, wherein the nephrotic syndrome comprises one or more immunoglobulin light chain deposits, such as one or more amyloid fibril deposits. Certain uses of the anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein are directed to methods of treating hepatomegaly in a subject in need thereof, comprising administering to the subject an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein, wherein the hepatomegaly comprises one or more immunoglobulin light chain deposits, such as one or more amyloid fibril deposits. Certain uses of the anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein are directed to methods of treating peripheral neuropathy in a subject in need thereof, comprising administering to the subject an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein, wherein the peripheral neuropathy comprises one or more immunoglobulin light chain deposits, such as one or more amyloid fibril deposits. Certain uses of the anti-human λ light chain antibodies or human λ light chain binding antibody fragments described herein are directed to methods of treating restrictive cardiomyopathy in a subject in need thereof, comprising administering to the subject an anti-human λ light chain antibody or human λ light chain binding antibody fragment described herein, wherein the restrictive cardiomyopathy comprises one or more immunoglobulin light chain deposits, such as one or more amyloid fibril deposits.

[0189] In some embodiments, administration of an anti-human λ light chain antibody or a human λ light chain binding antibody fragment to a subject results in a reduction and / or removal of one or more immunoglobulin light chain aggregates (e.g., amyloid fibrils) in the subject. In some embodiments, after administration of an anti-human λ light chain antibody or a human λ light chain binding antibody fragment, the size of immunoglobulin light chain aggregates (e.g., amyloid fibrils) in the subject is reduced.

[0190] In certain embodiments, the antibody can be administered to a subject in need by any route suitable for administering a pharmaceutical composition containing the antibody, such as subcutaneously, intraperitoneally, intravenously, intramuscularly, etc. In certain embodiments, the antibody is administered intravenously. In certain embodiments, the antibody is administered subcutaneously. In certain embodiments, the antibody is administered with a suitable dosage schedule, for example, once a week, twice a week, once a month, twice a month, once every two weeks, once every three weeks, or once a month, etc. In certain embodiments, the antibody is administered once every three weeks. The antibody can be administered in any therapeutically effective amount. In certain embodiments, the acceptable amount for treatment is about 0.1 mg / kg to about 50 mg / kg. In certain embodiments, the acceptable amount for treatment is about 1 mg / kg to about 40 mg / kg. In certain embodiments, the acceptable amount for treatment is about 1 mg / kg to about 20 mg / kg. In certain embodiments, the acceptable amount for treatment is about 1 mg / kg to about 10 mg / kg. In certain embodiments, the acceptable amount for treatment is about 5 mg / kg to about 30 mg / kg. In certain embodiments, the acceptable amount for treatment is about 5 mg / kg to about 20 mg / kg. A therapeutically effective amount includes an amount sufficient to ameliorate one or more symptoms associated with the disease or disorder being treated.

[0191] Pharmaceutically acceptable excipients, carriers and diluents

[0192] In certain embodiments, the anti-λ light chain antibodies of the present disclosure are included in pharmaceutical compositions comprising one or more pharmaceutically acceptable excipients, carriers, and diluents. Pharmaceutically acceptable excipients, carriers, and diluents may be included to increase the shelf life, stability, or applicability of the antibodies. Such compounds include salts, pH buffers, detergents, anticoagulants, and preservatives. In certain embodiments, the antibodies of the present disclosure are suspended in a sterile solution for administration. In certain embodiments, the solution comprises about 0.9% NaCl. In certain embodiments, the solution comprises about 5.0% dextrose. In certain embodiments, the solution further comprises one or more of: a buffer such as acetate, citrate, histidine, succinate, phosphate, bicarbonate, and hydroxymethylaminomethane (Tris); a surfactant such as polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), and poloxamer 188; a polyol / disaccharide / polysaccharide such as glucose, dextrose, mannose, mannitol, sorbitol, sucrose, trehalose, and dextran 40; an amino acid such as glycine or arginine; an antioxidant such as ascorbic acid, methionine; or a chelating agent such as EDTA or EGTA.

[0193] In certain embodiments, the antibodies of the present disclosure can be transported / stored through freeze drying and reconstructed before administration. In certain embodiments, the freeze-dried antibody formulation comprises a swelling agent, such as mannitol, sorbitol, sucrose, trehalose, dextran 40 or a combination thereof. The freeze-dried formulation can be contained in a vial consisting of glass or other suitable non-reactive materials. When the antibody is formulated, whether recombinant or not, it can be buffered with a certain pH, generally less than 7.0. In certain embodiments, the pH can be between 4.5 and 7.0, 4.5 and 6.5, 4.5 and 6.0, 4.5 and 5.5, 4.5 and 5.0 or 5.0 and 6.0.

[0194] Also described herein are kits comprising, in a suitable container, one or more antibodies described herein and one or more additional components selected from the group consisting of: instructions for use; diluents, excipients, carriers, and administration devices.

[0195] In certain embodiments, described herein are methods of preparing a lambda light chain amyloidosis treatment comprising mixing one or more pharmaceutically acceptable excipients, carriers, or diluents with an antibody of the disclosure. In certain embodiments, described herein are methods of preparing a cancer treatment for storage or transport comprising lyophilizing one or more antibodies of the disclosure.

[0196] definition

[0197] Antibodies are used in the broadest sense and generally encompass and / or refer to various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments, as long as they exhibit the desired antigen binding activity. In some embodiments, antibodies include complete antibodies and functional (antigen binding) antibody fragments, including fragment antigen binding (Fab) fragments, F(ab')2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, single-chain antibody fragments, including single-chain variable fragments (sFv or scFv) and single domain antibody (e.g., sdAb, sdFv, nanobody) fragments. In some embodiments, antibodies include genetically engineered and / or other modified forms of immunoglobulins, such as intracellular antibodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies and heterologous conjugate antibodies, multispecific antibodies, such as bispecific, antibodies, bispecific antibodies, triabodies and tetrabodies, tandem double scFv, tandem three scFv. In some embodiments, antibodies cover functional antibody fragments. In some embodiments, antibodies cover complete or full-length antibodies, including antibodies of any class or subclass, including IgG and its subclasses, IgM, IgE, IgA and IgD. The antibody may include a human IgG1 constant region. The antibody may include a human IgG4 constant region. In some embodiments, antibodies include but are not limited to full length and natural antibodies, as well as fragments and parts retaining their binding specificity, such as any specific binding portion thereof, including those with any number of immunoglobulin classes and / or isotypes (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA, IgD, IgE and IgM); and biologically relevant (antigen binding) fragments or their specific binding portions, including but not limited to Fab, F (ab') 2, Fv and scFv (single chain or related entities). Monoclonal antibodies are generally one of substantially homogeneous antibody compositions; therefore, any single antibody contained in a monoclonal antibody composition is identical, except for natural mutations that may be present in a small amount in the composition. The monoclonal antibody may comprise a human IgG1 constant region or a human IgG4 constant region.

[0198] Natural antibodies generally encompass and / or refer to naturally occurring immunoglobulin molecules with various structures. For example, natural IgG antibodies are heterotetrameric glycoproteins of about 150,000 daltons, consisting of two identical light chains and two disulfide-bonded identical heavy chains. From N-terminal to C-terminal, each heavy chain has a variable region (VH), also referred to as a variable heavy domain or a heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3). Similarly, from N-terminal to C-terminal, each light chain has a variable region (VL), also referred to as a variable light domain or a light chain variable domain, followed by a constant light (CL) domain. The light chain of an antibody can be assigned to one of the two types referred to as κ (κ) and λ (λ) based on the amino acid sequence of its constant domain.

[0199] Full length antibody, intact antibody and whole antibody are interchangeable and generally include and / or refer to antibodies having a structure substantially similar to a native antibody structure or having heavy chains that contain an Fc region as defined herein.

[0200] The human consensus framework generally encompasses and / or refers to the framework representing the most common amino acid residues in the selection of human immunoglobulin VL or VH framework sequences. In general, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. In general, the subgroup of sequences is such as Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda MD (1991), subgroups in Volumes 1-3. In some embodiments, for VL, the subgroup is such as Kabat et al., subgroup κI, supra. In some embodiments, for VH, the subgroup is such as Kabat et al., subgroup III, supra. Humanized antibodies encompass and refer to chimeric antibodies comprising amino acid residues from non-human HVR and amino acid residues from human framework regions (FR). In some embodiments, a humanized antibody will comprise substantially all of at least one and usually two variable domains, in which all or substantially all HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all FRs correspond to those of a human antibody. A humanized antibody optionally encompasses at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, such as a non-human antibody, refers to an antibody that has undergone humanization.

[0201] Complementarity determining regions or CDRs, synonymous with hypervariable regions or HVRs, generally include and / or refer to regions of an antibody that are hypervariable in sequence and / or form structurally defined loops (hypervariable loops) and / or contain antigen contact residues (antigen contacts). In some embodiments, complementarity determining regions or CDRs generally include and refer to non-contiguous amino acid sequences within an antibody variable region that confer antigen specificity and / or binding affinity. In general, there are three CDRs (CDR-H1, CDR-H2, CDR-H3) in each heavy chain variable region, and three CDRs (CDR-L1, CDR-L2, CDR-L3) in each light chain variable region.

[0202] Framework region or FR generally includes and / or refers to the non-CDR portion of the heavy chain and light chain variable region. In general, there are four FRs (FR-H1, FR-H2, FR-H3 and FR-H4) in each full-length heavy chain variable region, and four FRs (FR-L1, FR-L2, FR-L3 and FR-L4) in each full-length light chain variable region. In some embodiments, the framework region is defined by the non-CDR sequences of the variable region sequences.The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding site topography," J. Mol. Biol. 262, 732-745. ("Contact" numbering scheme); Lefranc MP et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003 Jan; 27 (1): 55-77 ("IMGT" numbering scheme); Honegger A and Plückthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun 8; 309 (3): 657-70, ("Aho" numbering scheme); and Whitelegg NR and Rees AR, "WAM: an improved algorithm for modelling antibodies on the WEB," Protein Eng. 2000 Dec; 13 (12): 819-24 ("AbM" numbering scheme). In certain embodiments, the CDRs of the antibodies described herein can be defined by a method selected from Kabat, Chothia, IMGT, Aho, AbM, or a combination thereof. In some embodiments, CDRs and FRs are defined by and / or according to the Kabat numbering scheme.In some embodiments, CDRs and FRs are defined by and / or according to the Chothia numbering scheme. In some embodiments, CDRs and FRs are defined by and / or according to the IMGT numbering scheme. In some embodiments, CDRs and FRs are defined by and / or according to the EU numbering scheme.

[0203] In some cases, the boundary of a given CDR or FR depends on the scheme used to identify and changes. For example, the Kabat scheme is based on structural comparisons, and the Chothia scheme is based on structural information. The numbering of both Kabat and Chothia schemes is based on the most common antibody region sequence length, and insertion is accommodated by inserting letters (e.g., "30a"), and disappearance occurs in some antibodies. These two schemes place certain insertions and deletions ("insertion and deletion (indel)") in different positions, resulting in different numbering. The Contact scheme is based on the analysis of complex crystal structures, and is similar to the Chothia numbering scheme in many respects.

[0204] Fc region generally encompasses and / or refers to the C-terminal region of an immunoglobulin heavy chain, which contains at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In some embodiments, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) in the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is carried out according to the EU numbering system (also referred to as the EU index), as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0205] Human antibodies generally encompass and / or refer to antibodies having an amino acid sequence that corresponds to the amino acid sequence of an antibody produced by a human or human cell or derived from a non-human source utilizing a human antibody library or other human antibody encoding sequences. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen binding residues.

[0206] The acceptor human framework region generally encompasses and / or refers to a framework comprising an amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human common framework, as defined below. The acceptor human framework "derived from" a human immunoglobulin framework or a human common framework may comprise the same amino acid sequence thereof or it may contain amino acid sequence changes. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less or 2 or less. In some embodiments, the VL acceptor human framework is identical in sequence to a VL human immunoglobulin framework sequence or a human common framework sequence.

[0207] Variable region or variable domain generally encompasses and / or refers to the domains of an antibody heavy chain or light chain that are involved in binding the antibody to an antigen. The variable domains of the heavy and light chains of a native antibody (V H and V L ) generally have a similar structure, each domain comprising four conserved framework regions (FRs) and three CDRs (see, e.g., Kindt et al., Kuby Immunology, 6th edition, WH Freeman and Co., p. 91 (2007)). A single V H or V L The domain may be sufficient to confer antigen binding specificity. H or V L The domain is used to separate antibodies that bind to a specific antigen from antibodies that bind to the antigen to screen for complementary V domains. L or V H Libraries of domains (see, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991)).

[0208] Affinity generally encompasses and / or refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, binding affinity generally encompasses and refers to intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (K D ) represents. Affinity can be measured by common methods known in the art, including the methods described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described throughout.

[0209] Affinity matured antibodies generally encompass and / or refer to antibodies with one or more alterations in one or more hypervariable regions (HVRs) which result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody that does not possess such alterations.

[0210] Binding and determination of binding (e.g., to an epitope comprising SEQ ID NO: 51) can be readily determined by methods known in the art (e.g., ELISA, surface plasmon resonance, biolayer interferometry, isothermal calorimetry, etc.). In some embodiments, binding is determined by ELISA. In some embodiments, binding comprises a K of less than, for example, 10^-5M (10uM). D , as measured by surface plasmon resonance, biolayer interferometry, or isothermal calorimetry. In some embodiments, the binding comprises a K of less than, for example, 10^-6 M (1 uM). D , surface plasmon resonance, biolayer interferometry, or isothermal calorimetry. In some embodiments, the combination includes a K of less than, for example, 10^-7 M (100 nM). D , surface plasmon resonance, biolayer interferometry or isothermal calorimetry.

[0211] Chimeric antibodies generally encompass and / or refer to antibodies in which a portion of the heavy chain and / or light chain is derived from a particular source or species, while the remainder of the heavy chain and / or light chain is derived from a different source or species. 1 The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and some of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are referred to as a, d, e, g, and m, respectively.

[0212] Monoclonal antibodies generally encompass and / or refer to antibodies obtained from a substantially homogeneous antibody population, that is, the single antibodies comprising the population are identical and / or bind to the same epitope, except for possible variant antibodies (e.g., variants containing naturally occurring mutations or occurring during the production of monoclonal antibody preparations), such variants are generally present in a small amount in the composition. Contrary to polyclonal antibody preparations that typically include different antibodies for different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed to a single determinant on an antigen. In some embodiments, the modifier monoclonal indicates that the antibody is obtained from a substantially homogeneous antibody population, and should not be understood as requiring antibodies to be produced by any particular method. For example, the monoclonal antibodies to be used in the present disclosure according to the composition disclosure can be prepared by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of human immunoglobulin loci, such methods for preparing monoclonal antibodies described herein, and other exemplary methods.

[0213] An isolated antibody generally encompasses and / or refers to an antibody separated from a component of its natural environment. In some embodiments, the antibody is purified to a purity greater than 95% or 99% as determined by, for example, electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reversed phase HPLC).

[0214] In certain embodiments, antibody comprises one or more naturally occurring amino acids. In certain embodiments, antibody is made up of naturally occurring amino acids. As used herein, naturally occurring amino acids include and / or refer to amino acids found in nature generally and not manipulated by humans. In some cases, naturally occurring includes and / or further refers to 20 kinds of conventional amino acids: alanine (A or Ala), cysteine ​​(C or Cys), aspartic acid (D or Asp), glutamic acid (E or Glu), phenylalanine (F or Phe), glycine (G or Gly), histidine (H or His), isoleucine (I or Ile), lysine (K or Lys), leucine (L or Leu), methionine (M or Met), asparagine (N or Asn), proline (P or Pro), glutamine (Q or Gln), arginine (R or Arg), serine (S or Ser), threonine (T or Thr), valine (V or Val), tryptophan (W or Trp) and tyrosine (Y or Tyr).

[0215] In some embodiments, the antibody comprises a variant sequence of an antibody. In some cases, amino acid substitutions may be made in the sequence of any antibody described herein without necessarily reducing or eliminating its activity (as measured by, for example, binding or functional assays described herein). Therefore, in some embodiments, the variant sequence comprises one or more amino acid substitutions (e.g., in the variable region or in one or more CDRs). In some embodiments, the variant sequence comprises one or more substitutions in one or more CDRs. In certain embodiments, the variant sequence comprises one amino acid substitution. In certain embodiments, the variant sequence comprises two amino acid substitutions. In certain embodiments, the variant sequence comprises three amino acid substitutions. In some cases, substitutions include conservative substitutions (e.g., substitutions with amino acids having comparable chemical features). In some cases, non-polar amino acids may be substituted and replaced with another non-polar amino acid, wherein non-polar amino acids include alanine, leucine, isoleucine, valine, glycine, proline, phenylalanine, tryptophan and methionine. In some cases, polar amino acids with a neutral charge may be substituted and replaced with another polar amino acid with a neutral charge, wherein polar amino acids with a neutral charge include serine, threonine, cysteine, tyrosine, asparagine and glutamine. In some cases, a positively charged amino acid can be substituted and replaced with another positively charged amino acid, wherein the positively charged amino acid includes arginine, lysine and histidine. In some cases, a negatively charged amino acid can be substituted and replaced with another negatively charged amino acid, wherein the negatively charged amino acid includes aspartic acid and glutamic acid. Examples of amino acid substitutions also include replacing L-amino acids with their corresponding D-amino acids, replacing cysteine ​​with homocysteine ​​or other non-natural amino acids.

[0216] In certain embodiments, the antibody comprises one or more non-natural amino acids. In certain embodiments, the antibody is composed of non-natural amino acids. As used herein, non-natural amino acids and / or non-natural amino acids include and / or refer to amino acid structures that cannot be biosynthetically produced in any organism using unmodified or modified genes from any organism. For example, these include but are not limited to modified amino acids and / or amino acid analogs that are not one of the 20 naturally occurring amino acids (e.g., non-natural side chain variant sequence amino acids), D-amino acids, high amino acids, β-high amino acids, N-methyl amino acids, α-methyl amino acids or. As further examples, unnatural amino acids also include 4-benzoylphenylalanine (Bpa), aminobenzoic acid (Abz), aminobutyric acid (Abu), aminocaproic acid (Ahx), aminoisobutyric acid (Aib), citrulline (Cit), diaminobutyric acid (Dab), diaminopropionic acid (Dap), diaminopropionic acid (Dap), γ-carboxyglutamate (Gla), homoalanine (Hala), homoarginine (Harg), homoasparagine (Hasn), homoaspartic acid (Hasp), homocysteine ​​(Hcys), homoglutamic acid (Hglu), homoglutamine (Hgln), homoisoleucine ( The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues. The non-natural amino acid residues may be substituted with the non-natural amino acid residues.

[0217] Stable formulations refer to and / or encompass formulations in which proteins (e.g., antibodies) are substantially kept physically and / or chemically stable and / or biologically active when stored at an expected storage temperature (e.g., 2-8°C). In some embodiments, the formulation substantially maintains its physical and chemical stability and its biological activity when stored. The storage period can generally be selected based on the expected shelf life of the formulation. In addition, the formulation is generally stable after the formulation is frozen (to, for example, -20°C) and thawed (e.g., after 1 or more freezing and thawing cycles). Various analytical techniques for measuring protein stability are available in the art, and are reviewed, for example, in Peptide and Protein Drug Delivery, 247-301, Vincent Lee, Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993). Stability can be measured at a selected temperature for a selected time period. Stability can be assessed qualitatively and / or quantitatively in a variety of ways, including assessing aggregate formation (e.g., using size exclusion chromatography, by measuring turbidity, and / or by visual inspection); assessing charge heterogeneity by using cation exchange chromatography or capillary zone electrophoresis; SDS-PAGE analysis to compare reduced and intact antibodies; assessing the biological activity or antigen binding function of the antibody; and the methods described herein. Instability can be related to any one or more of the following: aggregation, deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation), isomerization (e.g., Asp isomerization), shearing / hydrolysis / fragmentation (e.g., hinge region fragmentation), succinimide formation, unpaired cysteine, etc.

[0218] Pharmaceutically acceptable carriers generally encompass and / or refer to ingredients in pharmaceutical formulations other than active ingredients that are non-toxic to the subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0219] Polypeptide or protein are used interchangeably, and generally encompass and / or refer to a polymer of amino acid residues, and are not limited to a minimum length. The antibodies and antibody chains and other peptides provided, such as linkers and peptide-binding polypeptides, may include amino acid residues, including natural and / or non-natural amino acid residues. The term also includes post-expression modifications of the polypeptide, such as glycosylation, sialylation, acetylation, phosphorylation, etc. In some embodiments, the polypeptide may contain modifications relative to a natural or native sequence as long as the protein maintains the desired activity. These modifications may be deliberate, such as by site-directed mutagenesis, or may be accidental, such as by mutations of the host producing the protein or errors caused by PCR amplification.

[0220] The percentage of sequence identity (%) relative to the reference polypeptide sequence is the percentage of amino acid residues in the candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence after comparing the sequences and introducing gaps when necessary to achieve maximum percentage sequence identity, and not considering any conservative substitutions as a part of sequence identity. For determining the comparison of the purpose of percentage amino acid sequence identity, it can be realized in various known ways (for example, using publicly available computer software (such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software)). Suitable parameters for comparing sequences can be determined, including the algorithm required for maximum comparison on the entire full length of the compared sequences. However, for the purpose of this article, % amino acid sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program is written by Genentech, Inc., and the source code has been submitted to the US Copyright Office, Washington DC, 20559, together with user documents, where it is registered with US Copyright Registration Number TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, Calif., or can be compiled from source code. The ALIGN-2 program should be compiled for use on a UNIX operating system (including digital UNIX V4.0D). All sequence comparison parameters are set by the ALIGN-2 program and are unchanged.

[0221] As used herein, the term individual, patient or subject generally includes and / or refers to an individual diagnosed with at least one disease, condition or state, suspected of having at least one disease, condition or state, or having a risk of developing at least one disease, condition or state, and the described compositions and methods are useful for treating at least one disease, condition or state. In certain embodiments, the individual is a mammal. In certain embodiments, the mammal is a mouse, rat, rabbit, dog, cat, horse, cattle, sheep, pig, goat, llama, alpaca or yak. ​​In certain embodiments, the individual is a person.

[0222] The polypeptide described herein can be encoded by nucleic acid. Nucleic acid is a polynucleotide comprising two or more nucleotide bases. In certain embodiments, nucleic acid is a component of a vector that can be used to transfer a polypeptide encoding a polynucleotide into a cell. As used herein, the term "vector" refers to a nucleic acid molecule that can transport another nucleic acid that has been connected to it. One type of vector is a genomic integrated vector or "integrated vector", which can be integrated into the chromosomal DNA of a host cell. Another type of vector is an "episome" vector, for example, a nucleic acid that can carry out extrachromosomal replication. The vector that can guide the expression of the gene operably connected to them is referred to as an "expression vector" in this article. Suitable vectors include plasmids, bacterial artificial chromosomes, yeast artificial chromosomes, viral vectors, etc. In the expression vector, the regulatory elements (such as promoters, enhancers, polyadenylation signals) used to control transcription can be derived from mammals, microorganisms, viruses or insect genes. The ability to replicate in the host (usually given by the replication origin) and the selection gene that promotes the recognition of transformants can be additionally incorporated. A vector derived from a virus can be used, and the virus is such as a slow virus, a retrovirus, an adenovirus, an adeno-associated virus, etc. Plasmid vectors can be linearized to integrate into a chromosomal location. In certain embodiments, the expression vector is a plasmid. In certain embodiments, the expression vector is a lentivirus, an adenovirus, or an adeno-associated virus. In certain embodiments, the expression vector is an adenovirus. In certain embodiments, the expression vector is an adeno-associated virus. In certain embodiments, the expression vector is a lentivirus.

[0223] As used herein, the terms "homology", "homology" or "homology percentage" used herein to describe an amino acid sequence or a nucleic acid sequence relative to a reference sequence can be determined using the formula described by Karlin and Altschul (Proc. Natl. Acad. Sci. USA 87:2264-2268, 1990, revised as Proc. Natl. Acad. Sci. USA 90:5873-5877, 1993). Such formulas are incorporated into the Basic Local Alignment Search Tool (BLAST) program of Altschul et al. (J. Mol. Biol. 215:403-410, 1990). The homology percentage of a sequence can be determined using the latest version of BLAST as of the date of filing of this application.

[0224] The nucleic acids encoding the antibodies described herein can be used to infect, transfect, transform or otherwise transfect cells suitable for the nucleic acids, thereby enabling the production of antibodies for commercial or therapeutic use. Standard cell lines and methods for producing antibodies from large-scale cell cultures are known in the art. See, for example, Li et al., "Cell culture processes for monoclonal antibody production." Mabs. 2010 Sep-Oct; 2(5): 466–477. In certain embodiments, the cells are eukaryotic cells. In certain embodiments, the eukaryotic cells are mammalian cells. In certain embodiments, the mammalian cells are cell lines that can be used to produce antibodies, are Chinese hamster ovary cells (CHO cells), NSO mouse myeloma cells or Cells. In certain embodiments, nucleic acids encoding antibodies are integrated into genomic sites of cells that can be used to produce antibodies. In certain embodiments, methods of making antibodies are described herein, comprising culturing cells containing nucleic acids encoding antibodies under in vitro conditions sufficient to allow production and secretion of the antibody.

[0225] In certain embodiments, a master cell bank is described herein, comprising: (a) a mammalian cell line comprising a nucleic acid encoding an antibody described herein integrated at a genomic location; and (b) a cryoprotectant. In certain embodiments, the cryoprotectant comprises glycerol or DMSO. In certain embodiments, the master cell bank comprises: (a) a CHO cell line comprising a nucleic acid encoding an antibody disclosed herein; and (b) a cryoprotectant. In certain embodiments, the cryoprotectant comprises glycerol or DMSO. In certain embodiments, the master cell bank is contained in a suitable vial or container capable of withstanding freezing by liquid nitrogen.

[0226] Also described herein is the method for making antibody described herein.Such method is included in hatching the cell or clone of the nucleic acid comprising the encoding antibody in cell culture medium under the condition that is enough to allow antibody expression and secretion, and further gathers the antibody in the crops from cell culture medium.Harvesting can further comprise one or more purification steps to remove living cells, cell debris, non-antibody protein or polypeptide, undesirable salt, buffer agent and culture medium components.In certain embodiments, additional purification steps comprise centrifugation, ultracentrifugation, protein A purification, protein G purification, protein A / G purification or protein L purification and / or ion exchange chromatography.

[0227] As used herein, "treat," "treatment," or "treating" refers to, for example, intentional intervention in a physiological disease state that results in a decrease in the severity of a disease or condition; shortens the duration of a disease process; ameliorates or eliminates one or more symptoms associated with a disease or condition; or provides a beneficial effect to a subject suffering from a disease or condition. Treatment does not require a cure for the underlying disease or condition.

[0228] A "therapeutically effective amount," "effective dose," "effective amount," or "therapeutically effective dose" of a drug or therapeutic agent is any amount of a drug that, when used alone or in combination with another therapeutic agent, protects a subject from the onset of disease or promotes regression of disease as evidenced by a decrease in disease severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or prevents impairment or disability due to disease affliction. The ability of a therapeutic agent to promote regression of disease can be assessed using a variety of methods known to skilled practitioners, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by measuring the activity of the agent in in vitro assays.

[0229] As used herein, "pharmaceutically acceptable" with respect to a carrier, "excipient," or "diluent" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. In some aspects, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound (i.e., antibody) may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound.

[0230] The pharmaceutical compounds described herein may include one or more pharmaceutically acceptable salts. "Pharmaceutically acceptable salts" refers to salts that retain the desired biological activity of the parent compound and do not produce any undesirable toxicological effects (see, for example, Berge, SM, et al. (1977) J. Pharm. Sci. 66: 1-19). Examples of such salts include acid addition salts and base addition salts. Acid addition salts include salts derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, etc., and salts derived from non-toxic organic acids such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, aromatic acids, aliphatic sulfonic acids and aromatic sulfonic acids. Base addition salts include salts derived from alkaline earth metals such as sodium, potassium, magnesium, calcium, etc., and salts derived from non-toxic organic amines such as N, N'-dibenzylethylenediamine, N-methylglucamine, chloroprocaine, choline, diethanolamine, ethylenediamine, procaine, etc.

[0231] As used herein, treatment or treatment generally includes and / or refers to a drug or other intervention scheme for obtaining a beneficial or desired result in a recipient. Beneficial or desired results include, but are not limited to, therapeutic benefit and / or preventive benefit. Therapeutic benefit may refer to eradication or improvement of symptoms or the underlying condition being treated. In addition, by eradicating or improving one or more physiological symptoms associated with the underlying condition, a therapeutic benefit may be achieved so that improvement is observed in the object, although the object may still suffer from the underlying condition. Preventive effects include delaying, preventing or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing down, stopping or reversing the progress of a disease or condition, or any combination thereof. For preventive benefit, an object with a risk of developing a specific disease or an object reporting one or more physiological symptoms of a disease may undergo treatment, even if the disease may not yet be diagnosed. Those skilled in the art will recognize that given a potential population of individuals for treatment, not all people will respond to treatment or have equivalent responses. These individuals are considered to be treated.

[0232] In some embodiments, processing misfolded lambda light chains and / or processing lambda light chain amyloid deposits or processing lambda light chain aggregates can be used interchangeably herein. In some embodiments, processing misfolded lambda light chains includes treating a disorder characterized by misfolded lambda light chains. In some embodiments, processing misfolded lambda light chains includes reducing and / or reducing and / or suppressing lambda light chain aggregates (e.g., by measuring free circulating light chains) or its symptoms. In some embodiments, processing misfolded lambda light chains includes promoting monocytes and / or phagocytic cells to remove misfolded lambda light chains (e.g., by measuring free circulating light chains).

[0233] As used herein, monocytes or monocyte cells refer to and / or encompass a type of leukocyte or white blood cell. Monocytes are the largest class of leukocytes and can differentiate into, for example, macrophages. As part of the innate immune system of vertebrates, monocytes also affect the process of adaptive immunity. Monocytes can generally be divided into three types of monocytes in human blood: 1) Classical monocytes can be characterized by high levels of expression of the CD14 cell surface receptor (CD14 ++ CD16 - monocytes); 2) non-classical monocytes show low-level expression of CD14 and additional co-expression of the CD16 receptor (CD14 + CD16 ++ monocytes); and 3) intermediate monocytes with high expression of CD14 and low expression of CD16 (CD14++ CD16 + monocytes).

[0234] As used herein, macrophages generally refer to and / or encompass a type of white blood cell of the immune system that engulfs and digests cellular debris, foreign matter, microorganisms, cancer cells, and any other material that does not have a protein type specific to healthy body cells on its surface in a process called phagocytosis. In addition to increasing inflammation and stimulating the immune system, macrophages can also play an important anti-inflammatory role and can reduce immune responses by releasing cytokines.

[0235] As used herein, phagocytes generally refer to and / or encompass cells that are capable of phagocytosis and include both professional and non-professional phagocytes.

[0236] As used herein, amyloidosis generally refers to and / or encompasses diseases or conditions characterized by the misfolding of free immunoglobulin light chains (LC) (e.g., λLC), which can be deposited in the heart, kidney or liver as light chain amyloid (AL). This situation is usually caused by potential plasma cell dyscrasias, which cause plasma cells to secrete a large amount of excessive free immunoglobulin kappa (κ) and / or lambda (λ) light chains into the circulation. In some cases, the immunoglobulin kappa (κ) and / or lambda (λ) light chains in the circulation are misfolded, and primary amyloidosis or AL amyloidosis that is easy to aggregate and / or form amyloid deposits or aggregates generally refers to and / or encompasses the most common systemic amyloidosis types, with an incidence of 9-14 people per million per year in the Western world.

[0237] As used herein, plasma cell dyscrasias generally refer to and / or encompass proliferative plasma cell disorders, in which a single abnormal plasma cell clone (terminally differentiated B cell) undergoes unregulated amplification and produces light chain proteins (e.g., misfolded LC proteins) higher than normal amounts. Plasma cell dyscrasias can be divided into benign (as seen in monoclonal gammopathy of unknown significance (MGUS)) or malignant (as in multiple myeloma (MM) and plasma cell leukemia (PCL)). Regardless of the type of dyscrasias involved, these secreted light chains can undergo misfolding events, making them susceptible to aggregation and accumulation as amyloid in vital organs including the heart, kidney, and nervous system. This infiltration can lead to nephrotic syndrome, hepatomegaly, peripheral neuropathy, restrictive cardiomyopathy, and ultimately death.

[0238] As used herein, the words comprising (and any form of comprising, such as comprise and comprise), having (and any form of having, such as have and has), including (and any form of including, such as include and includes), or containing (and any form of containing, such as contain and contain) are inclusive or open-ended, and do not exclude additional, uncited elements or processing steps. Also as used herein, in any case or embodiment described herein, comprising can be replaced with essentially consisting of and / or consisting of. In any case or embodiment described herein, comprising as used herein can be replaced with essentially consisting of and / or consisting of.

[0239] As used herein, the term about in the context of a given value or range includes and / or refers to a value or range that is within 20%, within 10%, and / or within 5% of a given value or range.

[0240] As used herein, the term and / or will be considered as a specific disclosure of each of the two specified features or components, with or without the other. For example, A and / or B will be considered as a specific disclosure of each of (i) A, (ii) B, and (iii) A and B, just as if each were listed separately herein.

[0241] Example

[0242] The following illustrative examples represent embodiments of the compositions and methods described herein and are not meant to be limiting in any way.

[0243] Example 1 - Anti-human lambda light chain antibody

[0244] The problem that is at least partially solved is to create conformation-specific antibodies that bind to toxic light chains deposited in organs or their misfolded precursors in circulation while remaining out of contact with well-folded circulating light chains. Solving this problem provides a solution for the efficient direct treatment of pathogenic substances while avoiding interaction with free light chains that are usually present in large quantities and non-pathologically folded, which otherwise can serve as biological sinks or cause unwanted immune side effects in the middle of therapy. Without being limited by theory, the combination of antibodies and misfolded substances targets them for removal via antibody-dependent cellular phagocytosis (ADCP) mechanisms.

[0245] The light chain is composed of the variable domain (V L, AA1-108) and the constant domain (C L , AA109-214) composed of ~ 214 amino acids. The variable domain contains a hypervariable region composed of different amino acid sequences in each patient, while the constant domain shares sequence identity in all patients with the same isotype (κ or λ). It is believed that in each patient, the variable domain sequence of their unique light chain leads to the unique properties of the protein, such as amyloidosis tendency or amyloid structure, and many research efforts have been made to target this domain.

[0246] There are currently two competing products that aim to treat AL amyloidosis using an antibody-based amyloid depletion mechanism: Caelum CAEL-101 (Phase III in progress) and NEOD001 (Phase III, failure). Both companies have used a strategy to create a single antibody against the variable domain for the treatment of κ-AL amyloidosis and λ-AL amyloidosis patient populations. Although these antibodies are said to be effective for all patients (κ and λ), it is apparent in preclinical and clinical studies that the target engagement of these antibodies is inconsistent between individuals, as demonstrated by immunohistochemistry and clinical response measurements. This inconsistency can be attributed to the epitopes CAEL-101 and NEOD001. NEOD001 was originally designed to target serum amyloid A, but was found to have cross-reactivity to the light chain via interaction with a diamino acid epitope on the variable domain of the light chain protein. CAEL-101 is produced for epitopes in κAL patients. Recent cryo-EM structures of λ light chain amyloid extracted from patients show that the epitopes targeted by CAEL-101 and NEOD001 can be buried in the core of amyloid fibrils in a patient's light chain (e.g., inaccessible to the antibody) but accessible in another patient. This can lead to significant differences in binding affinity and clinical efficacy between individuals.

[0247] The antibody design method for generating anti-human λ light chain antibodies is to identify misfolded specific epitopes on the invariant constant domains, which will result in antibodies that bind to all variants of the same isotype with similar affinity. Laser capture microdissection mass spectrometry (LC-MS) studies show that the constant domains of the light chains (not just the variable domains) and full-length light chains or constant domain fragments are co-deposited in affected tissues, making them potential targets for antibody therapies that utilize amyloid protein depletion mechanisms. Therefore, κ and λ light chains can also be considered as two different targets.

[0248] The anti-human lambda light chain antibodies described herein were generated to bind to the misfolding specific epitope NKYAASSYLSL (SEQ ID NO: 51) on the lambda light chain protein. To generate misfolding specific antibodies, mice were immunized with immunogens displaying misfolding specific epitopes, and splenocytes were subsequently harvested and murine hybridoma cells were generated using established methods. Thirteen clones were identified and purified from mouse ascites for subsequent screening for desired misfolding specific properties.

[0249] Antibodies bound to denatured light chains

[0250] Initially, each of the thirteen clones was tested for its ability to bind to unfolded lambda light chains using Western blot to determine if these antibodies could recognize our target epitope in the context of full-length unfolded lambda light chains. Pooled lambda light chains from sera of multiple myeloma patients were used in the binding assays. For comparison, unfolded pooled kappa immunoglobulin light chains were included as well as whole serum from patients with confirmed lambda light chain (AL) amyloidosis. Figure 1 Data showing that five candidate antibodies successfully detected denatured (SDS-unfolded) lambda light chain proteins as well as one or more bands in patient serum samples are shown. The clones (clones 1-5) all showed strong binding to denatured light chains. Clones 4-5 also appeared to cross-react with denatured kappa light chains. Not all thirteen clones showed binding to unfolded lambda light chains. For the clones identified:

[0251] Clone 1 comprises CDRs identified / derived from a heavy chain variable domain comprising SEQ ID NO:1 and a light chain variable domain comprising SEQ ID NO:2.

[0252] Clone 2 comprises CDRs identified / derived from a heavy chain variable domain comprising SEQ ID NO:11 and a light chain variable domain comprising SEQ ID NO:12.

[0253] Clone 3 comprises CDRs identified / derived from a heavy chain variable domain comprising SEQ ID NO:21 and a light chain variable domain comprising SEQ ID NO:22.

[0254] Clone 4 comprises CDRs identified / derived from a heavy chain variable domain comprising SEQ ID NO:31 and a light chain variable domain comprising SEQ ID NO:32.

[0255] Clone 5 comprises CDRs identified / derived from a heavy chain variable domain comprising SEQ ID NO:41 and a light chain variable domain comprising SEQ ID NO:42.

[0256] Preferential binding to misfolded compared to native conformation

[0257] In five kinds of denatured light chain binders, it was studied whether these candidate monoclonal antibodies can distinguish and specifically bind to non-native λ light chain proteins, and do not bind to the native (well-folded) form. Wil, a representative λ light chain clone, was selected. Wil is a long-studied λ light chain variant first identified and sequenced in individuals with light chain amyloidosis (Pokkuluri et al., 1999). Full-length native Wil was produced in vitro by recombinant expression using established methods. When expressed and purified under these conditions, Wil forms stable, native-like light chain dimers. Then, by treating the native protein with urea and a reducing agent, alkylation is subsequently performed to permanently destroy disulfide bond formation to produce soluble, misfolded / unfolded Wil.

[0258] An indirect ELISA assay was performed to compare the binding of each antibody clone to native Wil relative to misfolded / unfolded Wil. A 96-well immunoassay plate was coated with a range of amounts of native or misfolded / unfolded Wil (0 to 800 ng protein / well) and each well was probed with our candidate antibodies or a commercial control. Figure 2A As illustrated in Figure 5, the bound antibody was detected using an HRP-conjugated secondary antibody and standard TMB chemistry. Figure 2A As shown in Figure 2B, clones 1 and 2 each showed high total signal, but also bound significantly to native Wil (although less than misfolded / unfolded Wil). Figure 2C As shown, clone 3 showed a slightly weaker signal but was slightly better at distinguishing between native and misfolded / unfolded Wil. Figure 2D -E indicates that clone 4 ( Figure 2D ) and clone 5 ( Figure 2E ) showed the weakest overall signal in this assay format, but also showed high specificity for misfolded Wil. Figure 2F All five clones demonstrated the ability to discriminate between misfolded / unfolded and native lambda light chains to some extent compared to the pan-specific commercial antibodies shown in Figure 3.

[0259] Preferential binding to aggregated / amyloid light chains compared to native conformation

[0260] The five clones were examined for their ability to bind to aggregated forms of lambda light chains (amyloid, pathogenic) using native Western blotting, as in Figure 3As exemplified in . The formation of Wil aggregates and amyloid was induced by prolonged oscillation at elevated temperatures using a previously established method, and amyloid formation was verified using the amyloid-sensitive dye Thioflavin T. Native protein blotting under "mild", low detergent conditions designed to preserve the folding of protein samples was performed, and the binding of our mAbs to native dimer Wil; misfolded / unfolded Wil; and aggregated / amyloid Wil was compared.

[0261] All candidate mAbs detected both misfolded / unfolded and amyloid / aggregated Wil. However, clones 1 and 2 showed the highest signal for the misfolded form of Wil. Despite the presence of staining of the native band, the signal was significantly reduced. It was suspected that the protein blotting and indirect ELISA methods (which fix the "native" Wil protein to the surface) may lead to false positive binding to the native protein by inducing a certain degree of misfolding, which would not be detected by the weakly binding clones (clones 3-5). Therefore, a capture format ELISA was used to interrogate the binding behavior of clones 1 and 2. In this format, a goat anti-mouse Fc capture antibody was coated into the ELISA wells, and clones 1 or 2 were captured by the antibody, with their Fab aligned in the appropriate orientation. Then, biotinylated Wil in native or misfolded form was added to these wells, and binding was determined by streptavidin. As Figure 4A and 4B As shown in , under these native conditions where Wil is in solution phase, we observed minimal binding to native Wil and preferential binding to the misfolded form in both clones 1 and 2, indicating conformational specificity. Clone 1 ( Figure 5A ) and Clone 2 ( Figure 5B ) both have their affinities measured by biolayer interferometry (BLI) indicating K values ​​of 59 nM and 256 nM, respectively. D Based on the above data, clone 1 was selected as the lead candidate for subsequent characterization (hereafter referred to as muLX-97).

[0262] In vitro characterization of muLX-97

[0263] The binding affinity of muLX-97 to multiple λ light chain variants in misfolded and native conformations was then measured. In addition to Wil, two light chain variants sequenced from cardiac amyloid deposits in patients with light chain amyloidosis were selected. See Oberti L et al., Concurrent structural and biophysical traits linkwith immunoglobulin light chains amyloid propensity. Sci Rep. 2017 Dec 1; 7(1): 16809 and Baur J et al., Identification of AL proteins from 10λ-AL amyloidosis patients by mass spectrometry extracted from abdominal fat and heart tissue. Amyloid. 2023 Mar; 30(1) 27-37. Again, BLI was used to monitor the misfolded and native forms of Wil ( Fig. 6A and 6D )、H3( Figure 6B and Fig. 6E ) and FOR102( Figure 6C and Fig. 6F ). muLX-97 had binding affinities of 59 nM, 317 nM, and 65 nM for misfolded Wil, H3, and FOR102, respectively. In the native conformation of all variants, only weak binding was observed (limited BLI signal at a very high concentration of 1000 nM). This limited association may be due to the presence of small amounts of misfolded Wil contaminating the "native" sample. Overall, this demonstrates that the conformational specificity of muLX-97 can be generalized to all lambda light chain variants.

[0264] Next, the binding affinity of muLX-97 to the amyloid form of the lambda light chain was determined. The true affinity for the aggregate cannot be measured using previously described methods in which the antibody or target is immobilized, so microthermophoresis (MST) was used. In this method, a small spot in a capillary containing a fluorescently tagged antibody is illuminated with an infrared laser in the presence or absence of the target. Local heating causes the protein to migrate out of the heated spot, and the rate of the change in migration (due to antibody binding to the ligand) can be used to measure affinity. Using the NanoTemper Monolith, the binding of muLX-97 to the misfolded and amyloid form of Wil was quantified and the results are shown in Figure 7. As Fig. 7A As shown in Figure 2, the K of misfolded Wil Dis 61 nM, which is consistent with the results from ELISA ( Figure 4A ) and BLI( Fig. 6A ) matches the result, and Figure 7B The affinity for amyloid Wil was shown to be 87 nM. These results support the notion that muLX-97 binds with high specificity to both misfolded and amyloid forms of lambda light chains. The binding affinities of the lambda AL variants determined using the different methods are summarized in Table 1.

[0265] Table 1: Binding affinity (K) of muLX-97 (clone 1) to various lambda light chain variants and conformations assessed by various biophysical and immunoassays. D ).

[0266]

[0267] Example 2 - Anti-human lambda light chain antibodies bind to human tissue amyloid

[0268] Ex vivo validation of muLX-97 target engagement

[0269] Next we evaluated whether muLX-97 could engage disease-associated targets in biopsy-confirmed lambda patient tissue. IHC + Congo red co-staining of several cases of patients with lambda light chain amyloidosis was performed. Congo red is an amyloid-specific dye that can also be visualized by fluorescence microscopy. Two representative cases are shown as examples. In the gastrointestinal case, the IHC staining pattern ( Fig. 8A ; dark staining) and Congo red fluorescence pattern ( Figure 8C ; white on dark background) closely matched in distribution and morphology. Similarly, this same co-staining was demonstrated in the soft tissue case ( Figure 8B and 8D ). Co-staining results showed that muLX-97 staining morphology was consistent with amyloid and was therefore amyloid-specific in cases of biopsy-confirmed λAL amyloidosis.

[0270] Positive staining in 5 cases of biopsy-confirmed λAL was also shown in the gastrointestinal tract ( Fig. 9A and 9B ),kidney( Fig. 9C ), and soft tissue ( Fig.9D and 9E ) and in the pancreas of patients with islet amyloid polypeptide amyloidosis or transthyretin amyloidosis ( Fig.9F ) or cardiac tissue ( Figure 9GThese results suggest that the disease-specific epitopes described herein are likely universal targets present in all cases of λ AL amyloidosis and that antibodies raised against said epitopes, such as muLX-97, are specific and targeted, with very low risk of cross-reactivity to other tissues.

[0271] Example 3 - Anti-human λ light chain antibodies induce antibody-dependent phagocytosis of ADCPλAL amyloid protein

[0272] After confirming the binding affinity and target engagement of muLX-97, the ability of muLX-97 to elicit an immune response and result in immune-mediated clearance of amyloid via antibody-mediated cellular phagocytosis (ADCP) was tested in a cellular model. In this model, amyloid Wil was conjugated to the pH-sensitive dye pH-sensitive rhodamine (pHrodo) and incubated with cultured macrophages and varying concentrations of muLX-97 or isotype control antibodies. If phagocytic uptake occurs, amyloid will be shuttled to lysosomes, where a decrease in pH results in fluorescence of phagocytic cells.

[0273] The ability of muLX-97 to induce ADCP in mouse macrophages was evaluated using an ADCP model system (RAW 264.7 and J774.1 cell lines). 40,000 cells were incubated with 2 μM pHrodo-tagged Wil amyloid and 0.1–200 nM muLX-97 or isotype control antibody. In RAW264.7 murine macrophages, ADCP was significantly reduced with isotype control antibodies ( Fig. 10A ) compared to muLX-97 ( Fig. 10B ) significantly increased the proportion of pHrodo-positive cells incubated with 5% PHrodo-positive cells. In J744.1 cells, the phagocytosis effect was more pronounced (up to 80% of cells were pHrodo-positive) and at 7.4 nM antibody concentration was significantly higher than that of cells treated with isotype control ( Fig.11A ) compared to muLX-97 treated cells ( Fig. 11B ) in the enhanced phagocytosis.

[0274] Quantification of the percentage of pHrodo-positive cells as a function of antibody concentration in these 2 model systems indicated that muLX-97 could induce phagocytic uptake of λAL amyloid, with EC in RAW 264.7 cells 50 is ∼20 nM and the maximum signal is 50% ( Fig. 12A ), and EC in J774.1 cells 50 is ∼7 nM and the maximum signal is 80% ( Fig. 12B). In summary, this cellular model shows that muLX-97 significantly enhances macrophage-mediated clearance of λ AL amyloid upon binding in a concentration-dependent manner, and the potency of this effect occurs at substoichiometric concentrations of the antibody.

[0275] Although preferred embodiments in the disclosure of the present composition have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Without departing from the present disclosure, those skilled in the art will now appreciate multiple modifications, changes and substitutions. It should be understood that various alternatives to the embodiments described herein may be used to practice the present invention. The following claims are intended to define the scope of the embodiments disclosed herein, and thus encompass methods and structures and their equivalents within the scope of these claims.

[0276] All publications, patent applications, issued patents, and other documents mentioned in this specification are incorporated herein by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions contained in text incorporated by reference are excluded to the extent they conflict with definitions in this disclosure.

[0277] Sequence Listing

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

Claims

1. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 3; (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 4; (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 5; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO:6; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 7; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:

8.

2. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 1, comprising: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or A light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3.

3. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 2, wherein: The heavy chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO: 1; and / or The light chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:

2.

4. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 13; (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 14; (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 15; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 16; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 17; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:

18.

5. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 4, comprising: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or A light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3.

6. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 5, wherein: The heavy chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO: 11; and / or The light chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:

12.

7. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 23; (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 2 4; (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 25; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 26; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 27; and / or (f) a light chain complementarity determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:

28.

8. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 7, comprising: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or A light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3.

9. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 8, wherein: The heavy chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO: 21; and / or The light chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:

22.

10. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 33; (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 34; (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO: 35; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 36; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 37; and / or (f) a light chain complementarity determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:

38.

11. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 10, comprising: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or A light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3.

12. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 11, wherein: The heavy chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:31; and / or The light chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:

32.

13. An anti-human λ light chain antibody or a human λ light chain binding antibody fragment thereof, comprising: (a) a heavy chain complementarity determining region 1 (H-CDR1) comprising the amino acid sequence shown in SEQ ID NO: 43; (b) a heavy chain complementarity determining region 2 (H-CDR2) comprising the amino acid sequence shown in SEQ ID NO:44; (c) a heavy chain complementarity determining region 3 (H-CDR3) comprising the amino acid sequence shown in SEQ ID NO:45; (d) a light chain complementary determining region 1 (L-CDR1) comprising the amino acid sequence shown in SEQ ID NO:46; (e) a light chain complementary determining region 2 (L-CDR2) comprising the amino acid sequence shown in SEQ ID NO: 47; and / or (f) a light chain complementary determining region 3 (L-CDR3) comprising the amino acid sequence shown in SEQ ID NO:

48.

14. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 13, comprising: a heavy chain variable region comprising said H-CDR1, H-CDR2 and H-CDR3; and / or A light chain variable region comprising said L-CDR1, L-CDR2 and L-CDR3.

15. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to claim 14, wherein: The heavy chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:41; and / or The light chain variable region comprises an amino acid sequence that is at least about 80% identical to SEQ ID NO:

42.

16. An anti-human λ light chain antibody or a human λ light chain antibody fragment thereof, comprising: A heavy chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO: 1; and / or A light chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:

2.

17. An anti-human λ light chain antibody or a human λ light chain binding antibody fragment thereof, comprising: A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1; and / or A light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

2.

18. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO: 11; and / or A light chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:

12.

19. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 11; and / or A light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

12.

20. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO: 21; and / or A light chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:

22.

21. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 21; and / or A light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

22.

22. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:31; and / or A light chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:

32.

23. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 31; and / or A light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

32.

24. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:41; and / or A light chain variable region comprising an amino acid sequence that is at least about 90%, 95%, 97%, 98% or 99% identical to SEQ ID NO:

42.

25. An anti-human λ light chain antibody or a human λ light chain binding fragment thereof, comprising: A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 41; and / or A light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

42.

26. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to any one of claims 1 to 25, wherein the anti-human λ light chain antibody is a monoclonal antibody.

27. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to any one of claims 1 to 26, wherein the anti-human λ light chain antibody is a chimeric antibody, a humanized antibody or a human antibody.

28. An anti-human λ light chain antibody or human λ light chain binding fragment thereof as described in any one of claims 1 to 25, wherein the human λ light chain binding antibody fragment comprises Fab, Fab', F(ab')2, scFv, dsFv, ds-scFv, dimer, miniantibody or bispecific antibody.

29. The anti-human λ light chain antibody or human λ light chain binding fragment thereof according to any one of claims 1 to 25, wherein the anti-human λ light chain antibody or human λ light chain binding fragment thereof is an IgG antibody.

30. The anti-human λ light chain antibody or human λ light chain binding fragment thereof of claim 29, wherein the IgG antibody is of IgG1 isotype.

31. The anti-human λ light chain antibody or human λ light chain binding fragment thereof of claim 29, wherein the IgG antibody is of the IgG4 isotype.

32. The anti-human λ light chain antibody or human λ light chain binding fragment thereof of any one of claims 1 to 31, wherein the anti-human λ light chain antibody or human λ light chain binding fragment thereof binds to an epitope comprising or consisting of SEQ ID NO:

51.

33. The method of claim 32, wherein binding to the epitope comprising SEQ ID NO: 51 is measured by ELISA, surface plasmon resonance, biolayer interferometry, or isothermal calorimetry.

34. The anti-human λ light chain antibody or human λ light chain binding fragment thereof as described in any one of claims 1 to 33, wherein the anti-human λ light chain antibody or human λ light chain binding fragment thereof is coupled to a tag.

35. The anti-human λ light chain antibody or human λ light chain binding fragment thereof of claim 34, wherein the tag comprises a chemical tag, a biological tag, a fluorescent tag, a radioisotope tag, or any combination thereof.

36. A pharmaceutical composition comprising the anti-human λ light chain antibody or human λ light chain binding fragment thereof according to any one of claims 1 to 35 and a pharmaceutically acceptable excipient, diluent or carrier.

37. A pharmaceutical composition comprising one or more anti-human λ light chain antibodies or human λ light chain binding fragments thereof as described in any one of claims 1 to 35 and a pharmaceutically acceptable excipient, diluent or carrier.

38. A nucleic acid encoding the anti-human λ light chain antibody or human λ light chain binding fragment thereof according to any one of claims 1 to 33.

39. A host cell comprising a nucleic acid encoding the anti-human λ light chain antibody or human λ light chain binding fragment thereof as described in any one of claims 1 to 33.

40. A method for diagnosing amyloidosis in a sample, the method comprising contacting a biological sample obtained from a subject with the anti-human λ light chain antibody or human λ light chain binding fragment of any one of claims 1 to 35.

41. A method for detecting free light chain polypeptides in a sample, the method comprising contacting the biological sample with an anti-human λ light chain antibody or human λ light chain binding fragment thereof according to any one of claims 1 to 35.

42. A method for detecting amyloid deposits in a sample, the method comprising contacting the biological sample with an anti-human lambda light chain antibody or human lambda light chain binding fragment thereof as described in any one of claims 1 to 35.

43. The method of any one of claims 40 to 42, wherein the sample is from blood, serum, plasma, solid tissue, or any combination thereof.

44. The method of any one of claims 40 to 42, wherein the sample is from a subject.

45. A method for promoting clearance of λ light chain aggregates by monocytes, the method comprising contacting the λ light chain aggregates with an anti-human λ light chain antibody or a human λ light chain binding fragment thereof as described in any one of claims 1 to 35.

46. ​​A method for increasing clearance of lambda light chain aggregates, the method comprising contacting the lambda light chain aggregates with an anti-human lambda light chain antibody or human lambda light chain binding fragment thereof as described in any one of claims 1 to 35.

47. A method for reducing lambda light chain aggregates in a subject, the method comprising administering to the subject an anti-human lambda light chain antibody or human lambda light chain binding fragment thereof as described in any one of claims 1 to 35, thereby reducing the number of lambda light chain aggregates in the subject.

48. A method of treating a disease or condition characterized by lambda light chain aggregates, the method comprising administering to the subject an anti-human lambda light chain antibody or human lambda light chain binding fragment thereof as described in any one of claims 1 to 35, thereby reducing the number of lambda light chain aggregates in the subject.

49. The method of any one of claims 47 or 48, wherein the subject has a plasma cell disorder.

50. The method of claim 49, wherein the plasma cell disorder comprises plasma cell dyscrasia.

51. The method of claim 50, wherein the plasma cell dyscrasia is selected from the group consisting of monoclonal gammopathy of undetermined significance (MGUS), multiple myeloma (MM), plasma cell leukemia (PCL), and any combination thereof.

52. The method of any one of claims 47 or 48, wherein the subject has systemic amyloidosis.

53. The method of any one of claims 47 or 48, wherein the subject has AL amyloidosis.

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