| 摘要 |
Conformational isomers of modified versions of alpha-Synuclein (alphaSyn), a protein that is associated with Parkinson's disease, have been designed and produced. These conformational isomers are produced by introducing cysteines into the alpha-Synuclein and scrambling the disulfide bonds to form stable and immunogenic isomers. These isomers are generically referred to as X-isomers. X-alphaSyn is an X-isomer of alphaSyn. X-alphaSyn is generally more immunogenic than wt-alphaSyn. Two groups of X-alphaSyn have been produced. One group is 3-disulfide X-alphaSyn(3SS) produced by introducing 6 Cys mutations into the alphaSyn. The second group is 2-disulfide X-alphaSyn(2SS), produced by introducing 4 Cys mutations into the alpha-Syn. In each of these two groups of X-alphaSyn, Cys was introduced not only wt-alphaSyn, but also in two Parkinson Disease associated alphaSyn mutants, A30P-alphaSyn and A53T-alphaSyn. All six sets of X-alphaSyn exhibit enhanced aggregation as compared to wt-alphaSyn, and should therefore more much more immunogenic. It was discovered that refolding the proteins in a CuSO4 buffer greatly enhanced oxidation and therefore diversity of the refolded proteins. Production of antibodies to X-alphaSyn is useful because such antibodies should bind to wt-alphaSyn and thus be useful as a therapeutic or for detection of alphaSyn in subjects.
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