期刊
BMC BIOTECHNOLOGY
卷 19, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s12896-019-0589-4
关键词
A beta; ApoE; Immunogold; Surface plasmon resonance; SPR; Scanning electron microscopy; SEM; Fibrils; Morphology; Abeta
资金
- National Microscopy Infrastructure, NMI at the Umea Core Facility for Electron Microscopy [VR-RFI 2016-00968]
- Demensforbundet
- Insamlingsstiftelsen at Umea University
- Ahlen-stiftelsen
- Hjarnfonden
- FAMY-Norrbotten
- Olle Engkvist stiftelse [199-0469]
- Medical Faculty of Umea University
Background: Immunogold labeling in combination with transmission electron microscopy analysis is a technique frequently used to correlate high-resolution morphology studies with detailed information regarding localization of specific antigens. Although powerful, the methodology has limitations and it is frequently difficult to acquire a stringent system where unspecific low-affinity interactions are removed prior to analysis. Results: We here describe a combinatorial strategy where surface plasmon resonance and immunogold labeling are used followed by a direct analysis of the sensor-chip surface by scanning electron microscopy. Using this approach, we have probed the interaction between amyloid-beta fibrils, associated to Alzheimer's disease, and apolipoprotein E, a well-known ligand frequently found co-deposited to the fibrillar form of A beta in vivo. The results display a lateral binding of ApoE along the amyloid fibrils and illustrates how the gold-beads represent a good reporter of the binding. Conclusions: This approach exposes a technique with generic features which enables both a quantitative and a morphological evaluation of a ligand-receptor based system. The methodology mediates an advantage compared to traditional immunogold labeling since all washing steps can be monitored and where a high stringency can be maintained throughout the experiment.
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