Improvement of a low pH antigen-antibody dissociation procedure for ELISA measurement of circulating anti-Aβ antibodies |
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| Authors: | Qingyou Li Marcia Gordon Chuanhai Cao Kenneth E Ugen Dave Morgan |
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| Affiliation: | (1) Alzheimer's Research Laboratory, Department of MolecularPharmacology and Physiology, University of South Florida, Tampa, FL 33612-4799, USA;(2) Department of Molecular Medicine, University of South Florida, Tampa, FL 33612-4799, USA;(3) Byrd Institute for Alzheimer's Disease, University of South Florida, Tampa, FL 33612-4799, USA |
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| Abstract: | Background Prior work from our group found that acid dissociation (pH 2.5 incubation) of serum from APP transgenic mice vaccinated against Aβ increased the apparent anti-Aβ titers, suggesting antibody masking by antigen in the ELISA assay. Subsequently, we found that pH 2.5 incubation of serum from unvaccinated non-transgenic mice showed antibody binding to Aβ1–42, but no increase when other proteins, including shorter Aβ peptides, coated the ELISA plate. To investigate further the effects of low pH incubation on apparent anti-Aβ1–42 signals, we examined normal sera from nonTg unvaccinated mice, nonTg mice vaccinated with Aβ peptide (to produce authentic anti-Aβ antibodies) or a monoclonal antibody against Aβ (6E10) using competitive-inhibition ELISA and Aβ epitope mapping assays. In addition, we examined use of a less stringent low pH procedure at pH 3.5, to ascertain if it had the same effects as the pH 2.5 procedure. |
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