JPWO2020073121A5 - - Google Patents

Description

For example, adjuvants that can be used include endogenous adjuvants (such as lipopolysaccharide), which are typically components of killed or attenuated bacteria used as vaccines. Exogenous adjuvants are typically immunomodulatory agents that are non-covalently bound to antigens and formulated to enhance the host's immune response. Aluminum hydroxide, aluminum sulfate, and aluminum phosphate (collectively commonly referred to as alum) are routinely used as adjuvants. A wide range of exogenous adjuvants can provoke potent immune responses to immunogens. These include saponins such as Stimulons (QS21, Aquila, Worcester, Mass.), or ISCOMs and (immunostimulatory complexes) conjugated to membrane protein antigens (immunostimulatory complexes) and products derived therefrom such as ISCOMATRIX. granulated particles, mineral oil, killed mycobacteria, and pluronic polymers containing mineral oil, Freund's complete adjuvant, lipid A and bacterial products such as muramyl dipeptide (MDP) and lipopolysaccharide (LPS) , and liposomes. is included.

Claims (74)

A cyclic compound comprising an α-Syn peptide comprising at least three residues of EKTKEQ (SEQ ID NO: 1 ) , a TEQ, and a linker, wherein the linker is covalently attached to the peptide N-terminal and C-terminal residues. , a cyclic compound. 2. The cyclic compound of Claim 1, wherein said α-Syn peptide is selected from EKTK (SEQ ID NO:2), KTKE (SEQ ID NO:3), and TKEQ (SEQ ID NO:4). 2. The cyclic compound of claim 1, wherein the α-Syn peptide is or comprises EKT, KTK, or KEQ. Cyclic compound according to any one of claims 1 to 3, wherein the α-Syn peptide is selected from EKTK (SEQ ID NO: 2) and TKEQ (SEQ ID NO: 4). Cyclic compound according to any one of claims 1 to 4, wherein the linker comprises or consists of 1 to 8 amino acids and/or one or more functionalizable moieties. 6. A cyclic compound according to claim 5, wherein the linker amino acid is selected from A and G and/or said functionalizable moiety is C. 7. Any one of claims 1-6, wherein the linker comprises or consists of GGCGG (SEQ ID NO: 13), GCGGGG (SEQ ID NO: 12), GGGCGG (SEQ ID NO: 15), or GGGGCGG (SEQ ID NO: 16) The cyclic compound according to. The cyclic compound of claim 1, wherein the cyclic compound is selected from compounds listed in any one of Tables 2-4. the cyclic compound is cyclo(CGTKEQGGG) (SEQ ID NO: 57), cyclo(CGGTKEQGGG) (SEQ ID NO: 47), cyclo(CGGTKEQGG) (SEQ ID NO: 48), cyclo(CGGGEKTKGG) (SEQ ID NO: 10), and cyclo(CGGGGEKTKGG) 2. The cyclic compound of claim 1, selected from (SEQ ID NO: 5). An immunogen comprising a cyclic compound according to any one of claims 1-9. 11. The immunogen of claim 10, wherein said cyclic compound is conjugated to a carrier protein or immunogenicity enhancing agent and/or is a multiple antigenic peptide (MAP). 12. The immunogen of claim 11, wherein said carrier protein is bovine serum albumin (BSA) or said immunogenicity enhancing agent is keyhole limpet hemocyanin (KLH). An antibody that binds to an epitope in said α-Syn peptide in the cyclic compound of any one of claims 1 to 9 or in said α-Syn peptide in a misfolded oligomeric α-Syn polypeptide, An antibody optionally produced using an immunogen according to any one of claims 10-12 or a composition comprising said immunogen. 14. The antibody of claim 13, wherein said composition used to produce said antibody comprises an adjuvant. Said adjuvants are aluminum phosphate , aluminum hydroxide , aluminum sulfate, saponins (such as Stimulon®), particles made from saponins (ISCOMs (immunostimulatory complexes) conjugated to membrane protein antigens (immunostimulatory complexes)). stimulant complexes) and ISCOMATRIX), pluronic polymers including mineral oil, killed mycobacteria and mineral oil, complete Freund's adjuvant, bacterial products (muramyl dipeptide (MDP), lipopolysaccharide (LPS), lipid A, etc.), 15. The antibody of claim 14, which is a liposome, or an oil-in-water emulsion (including a squalene-based oil-in-water emulsion) . Claim 13, wherein said antibody selectively binds to the cyclic compound of any one of claims 1-9 relative to the corresponding linear compound and/or native α-Syn polypeptide. 16. The antibody of any one of -15. 17. Any one of claims 13-16, wherein said α - Syn peptide and/or epitope comprises or consists of EKTK (SEQ ID NO:2), KTKE (SEQ ID NO:3), or TKEQ (SEQ ID NO:4) The antibody according to the paragraph. at least 2-fold, 3-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 18. The antibody of any one of claims 16-17, which is 30 - fold, at least 40-fold, at least 50-fold, at least 100-fold, at least 500-fold, at least 1000-fold more selective. 19. The antibody of any one of claims 16-18 , wherein the antibody selectively binds to misfolded oligomeric α-Syn polypeptide relative to native α-Syn polypeptide. said antibody is at least 2-fold, 3-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold greater than the native α-Syn polypeptide 20. The antibody of claim 19 , which is at least 40-fold, at least 50-fold, at least 100-fold, at least 500-fold, at least 1000-fold more selective. Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
of claims 16-20 , wherein the complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 have amino acid sequences of SEQ ID NOs: 61-66, respectively. The antibody of any one of paragraphs . Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
of claims 16-20 , wherein the complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 have amino acid sequences of SEQ ID NOs: 67-72, respectively. The antibody of any one of paragraphs . Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
of claims 16-20 , wherein the complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 have amino acid sequences of SEQ ID NOs: 73-78, respectively. The antibody of any one of paragraphs . Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
The complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 have amino acid sequences of SEQ ID NOs: 79-81, 76, 77, and 84, respectively. , the antibody of any one of claims 16-20. Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
the complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 are amino acid sequences of SEQ ID NOs: 79, 80, 81, 76, 77, and 84, respectively; The antibody of any one of claims 16-20, having Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
the complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 have amino acid sequences of SEQ ID NOs: 91-94, 71, and 96, respectively; The antibody of any one of claims 16-20. Said antibody comprises a heavy chain variable region and/or a light chain variable region, said heavy chain variable region comprising complementarity determining regions CDR-H1, CDR-H2, and CDR-H3, said light chain variable region comprising , comprising the complementarity determining regions CDR-L1, CDR-L2, and CDR-L3;
the complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 are amino acid sequences of SEQ ID NOs: 180, 181, 182, 183, 77, and 184, respectively The antibody of any one of claims 16-20, having said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 133 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 133, wherein said CDR sequences are maintained; said light chain variable region comprises the amino acid sequence of SEQ ID NO: 134 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 134; 22. The antibody of claim 21, wherein the CDR sequences are maintained. said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 135 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 135, wherein said CDR sequences are maintained said light chain variable region comprises the amino acid sequence of SEQ ID NO: 136 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 136; 23. The antibody of claim 22, wherein the CDR sequences are maintained. said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 137 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 137, wherein said CDR sequences are maintained; said light chain variable region comprises the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 138; 24. The antibody of claim 23, wherein the CDR sequences are maintained. said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 139 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 139, wherein said CDR sequences are maintained said light chain variable region comprises the amino acid sequence of SEQ ID NO: 140 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 140, said 25. The antibody of claim 24, wherein the CDR sequences are maintained. said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 141 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 141, wherein said CDR sequences are maintained said light chain variable region comprises the amino acid sequence of SEQ ID NO: 142 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 142, said 26. The antibody of claim 25, wherein the CDR sequences are maintained. said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 143 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 143, wherein said CDR sequences are maintained; said light chain variable region comprises the amino acid sequence of SEQ ID NO: 144 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 144; 27. The antibody of claim 26, wherein the CDR sequences are maintained. said heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 190 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 190, wherein said CDR sequences are maintained said light chain variable region comprises the amino acid sequence of SEQ ID NO: 191 or an amino acid sequence having at least 80%, 90%, 95%, or 98% sequence identity to SEQ ID NO: 191; 28. The antibody of claim 27, wherein the CDR sequences are maintained. said antibody is a monoclonal antibody, immunoglobulin molecule, Fab, Fab', F(ab)2, F(ab')2, Fv, disulfide-bonded Fv, scFv, disulfide-bonded scFv, single chain antibody, single domain antibody , diabodies, dimers, minibodies, bispecific antibody fragments, chimeric antibodies, humanized antibodies, and polyclonal antibodies. An immunoconjugate comprising an antibody according to any one of claims 16-35 and a moiety such as a detectable label or particle, optionally a magnetic particle. encoding said amino acid residue of a cyclic compound or immunogen according to any one of claims 1-12 , or an antibody according to any one of claims 16-35 or claim 36 A nucleic acid comprising a nucleic acid sequence encoding an immune complex of a nucleic acid sequence having SEQ ID NOS : 97-102 , wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 , respectively ; 38. The nucleic acid of claim 37 , comprising a nucleic acid sequence having SEQ ID NOS : 103-108 , wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 , respectively ; 38. The nucleic acid of claim 37 , comprising a nucleic acid sequence having SEQ ID NOS: 109-114 , wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 , respectively ; 38. The nucleic acid of claim 37 , comprising SEQ ID NOs: 115-117, 112 , 113 wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 , respectively , and 120. The nucleic acid of claim 37 . SEQ ID NOs: 115 , 116, 123, 112 wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 , respectively , 113, and 120 . SEQ ID NOs: 127-130, 107, wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 , respectively ; and 38. The nucleic acid of claim 37 , comprising a nucleic acid sequence having 132 . the nucleic acid sequences having SEQ ID NOs: 185-188, 113, and 189, wherein said nucleic acids encode complementarity determining regions CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3, respectively; 38. The nucleic acid of claim 37, comprising The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 145;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 145, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 133; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 133, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 146;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 146, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 134; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 134, wherein said amino acid sequence of said CDR sequence is maintained
coded by
39. The nucleic acid of claim 38. The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 147;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 147, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 135; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 135, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 148;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 148, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 136; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 136, wherein said amino acid sequence of said CDR sequence is maintained
coded by
40. A nucleic acid according to claim 39. The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 149;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 149, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 137; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 137, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 150;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 150, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 138; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 138, wherein said amino acid sequence of said CDR sequence is maintained
coded by
41. The nucleic acid of claim 40. The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 151;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 151, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 139; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 139, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 152;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 152, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 140; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 140, wherein said amino acid sequence of said CDR sequence is maintained
coded by
42. The nucleic acid of claim 41. The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 153;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 153, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 141; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 141, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 154;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 154, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 142; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 142, wherein said amino acid sequence of said CDR sequence is maintained
coded by
43. The nucleic acid of claim 42. The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 155;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 155, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 143; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 143, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 156;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 156, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 144; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 144, wherein said amino acid sequence of said CDR sequence is maintained
coded by
44. The nucleic acid of claim 43. The heavy chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 192;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 192, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 190; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 190, wherein said amino acid sequence of said CDR sequence is maintained
coded by
wherein the light chain variable region is
a nucleic acid sequence comprising SEQ ID NO: 193;
a sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 193, wherein said amino acid sequences of said CDR regions are maintained;
a sequence encoding SEQ ID NO: 191; or
a sequence encoding an amino acid sequence having at least 80%, 90%, 95%, 98%, or 99% sequence identity to SEQ ID NO: 191, wherein said amino acid sequence of said CDR sequence is maintained
coded by
45. The nucleic acid of claim 44. A vector comprising a nucleic acid according to any one of claims 37-51 . 53. The vector of claim 52 , further comprising a signal sequence optionally selected from SEQ ID NOs: 157-164, 169-177, and 179. A recombinant cell expressing the antibody of any one of claims 16-32. 55. The recombinant cell of claim 54 , wherein said recombinant cell is a mammalian cell, optionally a hybridoma cell or a CHO cell. A cyclic compound, immunogen, antibody, immune complex, nucleic acid, vector, or recombinant cell according to any one of claims 1-55 , optionally according to any one of claims 1-55 . comprising one or more, optionally two or more, or three or more of the cyclic compounds, immunogens, antibodies, immune conjugates, nucleic acids, vectors, or recombinant cells of 57. The cyclic compound, immunogen, antibody, immune conjugate, nucleic acid, vector, recombinant cell or composition of any one of claims 1-56 , optionally in one or more reagents, particles or plates. Included with the kit. administering to a subject the cyclic compound or immunogen according to any one of claims 1 to 12, or a composition comprising the cyclic compound or the immunogen; Isolating cells expressing antibodies specific for the immunogen and optionally selecting and/or isolating one or more antibodies that selectively bind to the misfolded oligomeric α-Syn polypeptide A method of producing an antibody according to any one of claims 16 to 35 , comprising: An assay for determining whether a test sample contains a misfolded α-Syn polypeptide, said method comprising:
a. The test sample is combined with the antibody of any one of claims 16 to 35 or the immune complex comprising said antibody under conditions permitting the formation of antibody:misfolded α-Syn polypeptide complexes. contacting the body, optionally with the immunoconjugate of claim 36 ;
b. any antibody: detecting and/or quantifying the presence of misfolded α-Syn polypeptide complexes,
An assay wherein said presence of a detectable complex indicates that said test sample may contain misfolded α-Syn polypeptide. 60. The assay of claim 59 , wherein the amount of antibody:misfolded α-Syn polypeptide complex is quantified and/or compared to a control. 61. An assay according to claim 59 or 60 , wherein said test sample comprises blood and/or serum and/or plasma and/or brain tissue extract and/or CSF. The assay of any one of claims 59-61 , wherein said test sample is a human sample. The assay of any one of claims 59-62 , wherein detecting said complex comprises contacting said complex with a pan α-Syn antibody. said assay further comprising detecting and/or quantifying said presence of antibody:misfolded α-Syn polypeptide complexes in subsequent test samples and optionally comparing said test samples. , the assay of any one of claims 59-63 . The assay of any one of claims 59-64 , wherein said subject is being treated for alpha-synucleinopathy. The assay of any one of claims 59-65 , wherein said subject is untreated for alpha-synucleinopathy. An assay according to any one of claims 59 to 66 , wherein said antibody contacted with said test sample is conjugated to particles, optionally magnetic beads. 68. Any one of claims 59 to 67 , wherein said detecting said complex or quantifying said complex comprises contacting said complex with a labeled pan α-syn antibody. assay. 69. An assay according to any one of claims 59 to 68 for use in a method of diagnosing whether a subject has alpha-synucleinopathy,
a. detecting the amount of misfolded alpha-synuclein in a test sample of said subject;
b. comparing the amount of misfolded alpha-synuclein to a control, wherein the control is a cut-off or range found in a population of control samples;
if said amount of misfolded alpha-synuclein is higher than the level or range found in a normal control sample or within the range found in a control sample from a subject with said synucleinopathy; are likely to have an α-synucleinopathy. 70. The assay of claim 69 , wherein said alpha-synucleinopathy is selected from Parkinson's disease (PD), Lewy body disease (LBD), or multiple system atrophy. A method of inhibiting misfolded α - syn toxicity, comprising administering to a cell population or a subject in need thereof an effective amount of the antibody or immunoconjugate of any one of claims 13-36 , or administering a composition comprising said antibody or immunoconjugate, wherein said antibody is associated with misfolded oligomeric α-Syn compared to monomer, native tetramer, and/or insoluble fibrillar α-synuclein species. A method of selectively binding. 71. A method according to claim 71 , wherein said composition comprises one or more antibodies and/or immunoconjugates according to any one of claims 13-36 . administering an effective amount of the antibody of any one of claims 16-35 , the immunoconjugate of claim 36 , or the composition of claim 56 to a subject in need thereof wherein said antibody has an α-synucleinopathy that selectively binds to misfolded oligomeric α-synuclein relative to monomeric, native tetrameric, and/or insoluble fibrillar α-synuclein species 73. The method of claim 71 or 72 for treating a subject. 74. The method of any one of claims 71-73 , wherein said alpha-synucleinopathy is selected from Parkinson's disease (PD), Lewy body disease (LBD), or multiple system atrophy.