CONFORMATIONAL ISOMERS OF ALPHA-SYNUCLEIN, ANTIBODIES THERETO AND METHODS OF THEIR MANUFACTURE AND USE

• Format: Patent
• Language: English
• Published: 18.12.2008

4 Conformational isomers of modified versions of α-Synuclein (αSyn), a protein that is associated with Parkinson's disease, have been designed and produced. These conformational isomers are produced by introducing cysteines into the α-Synuclein and scrambling the disulfide bonds to form stable and immunogenic isomers. These isomers are generically referred to as X-isomers. X-αSyn is an X-isomer of αSyn. X-αSyn is generally more immunogenic than wt-αSyn. Two groups of X-αSyn have been produced. One group is 3-disulfide X-αSyn(3SS) produced by introducing 6 Cys mutations into the αSyn. The second group is 2-disulfide X-αSyn(2SS), produced by introducing 4 Cys mutations into the α-Syn. In each of these two groups of X-αSyn, Cys was introduced not only wt-αSyn, but also in two Parkinson Disease associated αSyn mutants, A30P-αSyn and A53T-αSyn. All six sets of X-αSyn exhibit enhanced aggregation as compared to wt-αSyn, and should therefore more much more immunogenic. It was discovered that refolding the proteins in a CuSObuffer greatly enhanced oxidation and therefore diversity of the refolded proteins. Production of antibodies to X-αSyn is useful because such antibodies should bind to wt-αSyn and thus be useful as a therapeutic or for detection of αSyn in subjects.