Morphological analysis of Apolipoprotein E binding to A beta Amyloid using a combination of Surface Plasmon Resonance, Immunogold Labeling and Scanning Electron Microscopy

Islam, Tohidul (author): Umeå universitet,Institutionen för medicinsk kemi och biofysik

Gharibyan, Anna L. (author): Umeå universitet,Institutionen för medicinsk kemi och biofysik

Olofsson, Anders, 1970- (author): Umeå universitet,Institutionen för medicinsk kemi och biofysik

(creator_code:org_t)

Related links:: https://doi.org/10.1...; show more...; https://umu.diva-por... (primary) (Raw object); https://bmcbiotechno...; https://urn.kb.se/re...; https://doi.org/10.1...; show less...

Abstract Subject headings

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.

By the author/editor: Islam, Tohidul; Gharibyan, Anna ...; Lee, Cheng Choo; Olofsson, Anders ...

About the subject

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