title: Specificity and sensitivity of the Abeta oligomer ELISA.
abstract: Abeta soluble oligomers are believed to play a key role in the development of Alzheimer's disease (AD). An enzyme-linked immunosorbent assay (ELISA) commonly used to measure these proteins uses the same monoclonal antibody as both capture and reporter antibody. The objective of this study was to examine the specificity and sensitivity of this procedure, using monoclonal anti-Abeta antibody 6E10 as capture antibody and biotinylated 6E10 as reporter antibody. At comparable concentrations of Abeta soluble oligomers and low molecular weight (LMW) Abeta peptides, optical density (OD) values were four- to five-fold higher for the oligomer preparation than for the LMW Abeta. The LMW Abeta preparation, when evaluated by western blots of gels run under native conditions, showed only one band even after storage at 4 °C for more than two months, suggesting that the ELISA was detecting Abeta monomer as well as Abeta oligomers. Possible explanations for these results are that (1) the LMW Abeta preparation may contain Abeta oligomer species below the limit of detection of western blot, but still detectable by ELISA, or (2) some nonspecific binding of the LMW Abeta to the ELISA plate may have occurred, allowing its relevant epitope to remain available for binding by the reporter antibody. Because of the possibility that this ELISA may not be oligomer-specific, it seems prudent to suggest that it should be used in combination with other methods, rather than as the sole technique, for measuring Abeta oligomers in biological specimens.
authors.affiliation: Department of Neurology Research, William Beaumont Hospital Research Institute, 3811 West Thirteen Mile Road, Suite 507, Royal Oak, MI 48073, USA. Andrea.Klaver@beaumont.edu
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Klaver
authors.forename: Lynnae M
authors.initials: LM
authors.lastname: Patrias
authors.forename: John M
authors.initials: JM
authors.lastname: Finke
authors.forename: David A
authors.initials: DA
authors.lastname: Loeffler
mesh_terms: D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000378:metabolism*; D000906:Antibodies / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D012680:Sensitivity and Specificity
chemical_list: D016229:Amyloid beta-Peptides; D000906:Antibodies; D010446:Peptide Fragments; C075222:amyloid beta-protein (1-42)