Antibodies and Vaccines for Use in Therapeutic and Diagnostic Methods for Alpha-Synuclein-Related Disorders

Abstract

Methods for detecting α-synuclein oligomers in vivo comprise administering an antibody to an individual suspected of carrying soluble α-synuclein, wherein the antibody is produced from a stabilized soluble α-synuclein oligomer and is capable of binding the stabilized soluble α-synuclein oligomer, wherein the stabilized soluble α-synuclein oligomer has a lower formation rate to a non-soluble aggregated form than a non-stabilized soluble oligomer of the α-synuclein, and wherein the antibody is labelled with a detectable marker; and detecting the presence of any complex formed between the antibody and soluble α-synuclein by detection of the marker.

1 . A method of detecting α-synuclein oligomers in vivo, comprising
 administering an antibody to an individual suspected of carrying soluble α-synuclein, wherein the antibody is produced from a stabilized soluble α-synuclein oligomer and is capable of binding the stabilized soluble α-synuclein oligomer, wherein the stabilized soluble α-synuclein oligomer has a lower formation rate to a non-soluble aggregated form than a non-stabilized soluble oligomer of the α-synuclein, and wherein the antibody is labelled with a detectable marker; and 
 detecting the presence of any complex formed between the antibody and soluble α-synuclein by detection of the marker. 
 
     
     
         2 . The method according to  claim 1 , wherein the stabilized soluble α-synuclein oligomer comprises a stabilized soluble α-synuclein protofibril. 
     
     
         3 . The method according to  claim 1 , wherein the antibody is monoclonal. 
     
     
         4 . The method according to  claim 1 , wherein the antibody is humanized or modified to reduce antigenicity in humans. 
     
     
         5 . The method according to  claim 1 , wherein the antibody is of IgG class. 
     
     
         6 . The method according to  claim 1 , wherein the antibody is a Fab fragment selected from F(ab), F(ab) 2 , and DiFabody. 
     
     
         7 . The method according to  claim 1 , wherein the antibody is polyclonal. 
     
     
         8 . The method according to  claim 1 , wherein the antibody is a single chain antibody selected from scFv-Fc and scFab. 
     
     
         9 . The method according to  claim 1 , wherein the antibody is administered in combination with one or more excipients selected from the group consisting of antibacterial agents, adjuvants, buffers, salts, pH-regulators, detergents, and any combination thereof, that are pharmaceutically acceptable for human or veterinary use. 
     
     
         10 . The method according to  claim 1 , wherein the stabilized soluble α-synuclein oligomer comprises a soluble α-synuclein oligomer modified with a hydrophobic organic agent comprising a) a compound selected from the group consisting of 4-hydroxy-2-nonenal, malondialdehyde, 4-oxo-2-nonenal, acrolein, and any combination thereof, b) a saturated, unsaturated, or polyunsaturated fatty acid, or any combination thereof, c) a non-ionic detergent, a zwitterionic detergent, or any combination thereof, d) at least one compound selected from the group consisting of triglycerides, phospholipids, sphingolipids, gangliosides, cholesterol, cholesterol-esters, and long chain alcohols, and any combination thereof, or e) a bile acid derivative selected from the group consisting of deoxycholate, cholate and taurocholate, and any combination thereof. 
     
     
         11 . The method according to  claim 1 , wherein the stabilized soluble α-synuclein oligomer comprises a soluble α-synuclein oligomer modified with 1-α-hydroxy-secosterol or a soluble α-synuclein oligomer cross-linked with at least one protein cross-linking agent selected from the group consisting of disuccinimidyl tartrate, bis-sulfosuccinimidyl suberimidate and 3,3-dithiobis-sulfosuccinimidyl propionate, and any combination thereof. 
     
     
         12 . The method according to  claim 1 , wherein the α-synuclein oligomer comprises wild type α-synuclein (SEQ ID NO: 1), wild type α-synuclein in which one or more Ser and/or Ala amino acids have been replaced with Cys via site-directed mutagenesis, a mutant selected from the group consisting of A30P (SEQ ID NO: 2), E46K (SEQ ID NO: 3) and A53T (SEQ ID NO: 4), or any combination thereof. 
     
     
         13 . The method according to  claim 1 , wherein the ratio of the IC 50  binding strength of the antibody to soluble α-synuclein protofibrils to the IC 50  binding strength of the antibody to α-synuclein monomers is in the range of about 1:50 to 1:2000. 
     
     
         14 . The method according to  claim 1 , wherein the ratio of the IC 50  binding strength of the antibody to soluble α-synuclein protofibrils to the IC 50  binding strength of the antibody to insoluble α-synuclein fibrils is in the range of about 1:2 to 1:2000. 
     
     
         15 . The method according to  claim 1 , wherein the stabilized soluble α-synuclein oligomer comprises a stabilized soluble α-synuclein protofibril, wherein the stabilized soluble α-synuclein protofibril comprises a soluble α-synuclein protofibril modified with a hydrophobic organic agent comprising a compound selected from the group consisting of 4-hydroxy-2-nonenal, malondialdehyde, 4-oxo-2-nonenal, acrolein, and any combination thereof, and wherein the antibody is monoclonal or comprises a fragment thereof. 
     
     
         16 . The method according to  claim 1 , wherein the antibody has been collected from a non-human animal to which stabilized soluble α-synuclein oligomer had been administered. 
     
     
         17 . The method according to  claim 1 , wherein the antibody has been produced by hybridoma technology, phage display, ribosome display, mammalian cell display or bacterial display. 
     
     
         18 . The method according to  claim 1 , wherein the stabilized soluble α-synuclein oligomer comprises a stabilized soluble α-synuclein protofibril, and wherein the antibody has been collected from a non-human animal to which the stabilized soluble α-synuclein protofibril had been administered. 
     
     
         19 . The method according to  claim 1 , wherein the stabilized soluble α-synuclein oligomer comprises a stabilized soluble α-synuclein protofibril, and wherein the antibody has been produced by hybridoma technology, phage display, ribosome display, mammalian cell display or bacterial display. 
     
     
         20 . The method according to  claim 1 , wherein the individual is suspected of having a neurodegenerative disorder with α-synuclein pathology characterized by deposition of Lewy bodies and Lewy neurites. 
     
     
         21 . The method according to  claim 1 , wherein the individual is suspected of having Parkinson's disease (PD), dementia with Lewy bodies (DLB), the Lewy body variant of Alzheimer's disease, or multiple system atrophy (MSA). 
     
     
         22 . The method according to  claim 1 , wherein the individual is undergoing treatment for a neurodegenerative disorder with α-synuclein pathology characterized by deposition of Lewy bodies and Lewy neurites. 
     
     
         23 . The method according to  claim 1 , wherein the detectable marker comprises a radioactive ligand. 
     
     
         24 . The method according to  claim 23 , wherein the radioactive ligand comprises  131 I,  14 C,  3 H or  58 Ga.