Sensitivity and specificity of phospho‐Ser129 α‐synuclein monoclonal antibodies

• DOI: 10.1002/cne.24468
• Publication Date: 2018-06-11T09:27:05Z
• Authors: Vedad Delic; Sidhanth Chandra; Hisham Abdelmotilib; Tyler Maltbie; Shijie Wang; Danielle Kem; Hunter J. Scott; Rachel N. Underwood; Zhiyong Liu; Laura A. Volpicel...; Andrew B. West
• Keywords: Antibodies, Monoclonal; Brain; Rats; 3. Good health; Mice, Inbred C57BL; Rats, Sprague-Dawley; Mice; 03 medical and health sciences; 0302 clinical medicine; Antibody Specificity; alpha-Synuclein; Animals

Abstract α‐Synuclein (α‐syn) is an abundant presynaptic protein that the primary constituent of inclusions define Lewy body diseases (LBDs). In these inclusions, α‐syn phosphorylated at serine‐129 residue. Antibodies directed to this phosphorylation site are used measure inclusion abundance and stage disease progression in preclinical models as well postmortem tissues LBDs. While it critical reliably identify phospho‐specific antibodies often cross‐react with nonspecific antigens. Four commercially available monoclonal antibodies, two from rabbits (clones EP1536Y MJF‐R13) mice (81a pSyn#64), have been most widely detecting pS129‐α‐syn inclusions. Here, we systematically evaluated brain sections lysates rats mice. All detected were induced by preformed fibrils wild‐type Antibody titrations revealed clones 81a comparably labeled both perikarya neuronal processes contrast MJF‐R13 pSyn#64 incompletely various antibody concentrations. Except for EP1536Y, produced diffuse neuropil labeling knockout injected monomeric α‐syn, some staining titrating By immunoblot, all cross‐reacted proteins other than warranting caution interpretations specificity. Clone uniquely robustly endogenous highly soluble fractions mouse brain. summary, had highest sensitivity specificity pS129‐α‐syn.