Generation and Partial Characterization of Rabbit Monoclonal Antibody to Pyroglutamate Amyloid-β3-42 (pE3-Aβ)
| Autor: | Thomas Wisniewski, Pankaj Mehta, Bruce A. Patrick, Sangita P. Mehta, David Miller, Bozena Mazur-Kolecka, Marc Barshatzky, Jerzy Wegiel, Janusz Frackowiak |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Genetically modified mouse Adult Amyloid medicine.drug_class Antibody Affinity Fluorescent Antibody Technique Peptide Enzyme-Linked Immunosorbent Assay Monoclonal antibody Epitope 03 medical and health sciences Immunolabeling 0302 clinical medicine Alzheimer Disease Antibody Specificity medicine Animals Humans Aged chemistry.chemical_classification Amyloid beta-Peptides biology General Neuroscience Antibodies Monoclonal General Medicine Middle Aged Molecular biology Isotype Peptide Fragments Frontal Lobe Psychiatry and Mental health Clinical Psychology 030104 developmental biology chemistry biology.protein Rabbits Geriatrics and Gerontology Antibody Down Syndrome 030217 neurology & neurosurgery Biomarkers Epitope Mapping |
| Zdroj: | Journal of Alzheimer's disease : JAD. 62(4) |
| ISSN: | 1875-8908 |
| Popis: | N-terminally truncated pyroglutamate amyloid-β (Aβ) peptide starting at position 3 represents a significant fraction of Aβ peptides (pE3-Aβ) in amyloid plaques of postmortem brains from patients with Alzheimer's disease (AD) and older persons with Down syndrome (DS). Studies in transgenic mouse models of AD also showed that pE3-Aβ is a major component of plaques, and mouse monoclonal antibody to pE3-Aβ appears to be a desirable therapeutic agent for AD. Since small peptides do not typically elicit a good immune response in mice, but do so favorably in rabbits, our aims were to generate and partially characterize a rabbit monoclonal antibody (RabmAb) to pE3-Aβ. The generated RabmAb was found to be specific for pE3-Aβ, since it showed no reactivity with Aβ16, Aβ40, Aβ42, Aβ3-11, and pE11-17 Aβ peptides in an enzyme linked immunosorbent assay (ELISA). The isotype of the antibody was found to be IgG class. The antibody possesses high affinity to pE3-Aβ with dissociation constant (KD) for the antibody of 1 nM. The epitope of the antibody lies within the sequence of pE3-FRHD. In dot blotting, the optimal detection of pE3-Aβ was at an antibody concentration of 0.5 μg/ml. The threshold of pE3-Aβ detection was 2 fmol. The antibody was sensitive enough to detect 10 pg/ml of pE3-Aβ in sandwich ELISA. pE3-Aβ was detected in AD and DS brain extracts in ELISA and immunoblotting. Immunohistological studies showed immunolabeling of plaques and blood vessels in brains from patients with AD, and DS showing AD pathology. Thus, the antibody can be widely applied in AD and DS research, and therapeutic applications. |
| Databáze: | OpenAIRE |
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