Sensitive and specific detection of α-synuclein in human plasma.
alpha-Synuclein (alphasyn) mutations, overexpression, misfolding, and aggregation are associated with Parkinson's disease. This protein has been intensively studied in neuronal systems. However, alphasyn is also present in extracellular fluids, such as cerebrospinal fluid and blood plasma. Recent studies have attempted to quantify its levels and compare these in various extracellular fluids of control and Parkinson's disease subjects. Data from these studies have been difficult to interpret, suggesting that more sensitive, standardized, and well-characterized assays of larger cohorts are required. Here, we describe the development of a new ELISA specifically for quantifying alphasyn in human plasma. An initial assay, using a commercial anti-alphasyn monoclonal antibody (211; Santa Cruz Biotechnology, Santa Cruz, CA) and based on a published protocol, was adapted for use in human plasma. In addition, we have developed a novel alphasyn-specific antibody for the assay that has very high sensitivity and signal:noise characteristics. Assays with either antibody showed high specificity for alphasyn, and detected it in a variety of sample types, including plasma. These assays can now be employed on large cohorts of patients and control subjects to determine whether plasma levels are altered in disease. Although measuring extracellular alphasyn levels may prove to be a useful biomarker of Parkinson's disease, it should also be a powerful tool for basic research aimed at understanding the normal and pathological physiology of alphasyn secretion. .
Tinsley
R B
RB
Florey Neuroscience Institutes, University of Melbourne, Melbourne, Victoria 3010, Australia. rogan.tinsley@florey.com.au
Kotschet
K
K
Modesto
D
D
Ng
H
H
Wang
Y
Y
Nagley
P
P
Shaw
G
G
Horne
M K
MK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood / Q000378:metabolism; D001666:Binding Sites, Antibody; D015415:Biomarkers / Q000097:blood / Q000134:cerebrospinal fluid / Q000378:metabolism; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D010300:Parkinson Disease / Q000097:blood* / Q000175:diagnosis*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D013045:Species Specificity; D051844:alpha-Synuclein / Q000097:blood* / Q000134:cerebrospinal fluid / Q000235:genetics
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D051844:alpha-Synuclein