Year: 2026
Type: preprint
Abstract: Affinity graphene grids provide a promising approach for selective protein capture in cryo-EM. Here, we introduce a split-GFP graphene grid platform(SGFP-Grid), in which graphene-conjugated GFP1–10 selectively captures GFP11-tagged proteins from low-concentration samples or cell lysates. This platform enables rapid assessment of target protein enrichment and particle distribution before vitrification via fluorescence imaging, while the grid design positions captured proteins away from the graphene surface and air–water interface. We also introduce a unique strategy to minimize nonspecific protein adsorption, thereby improving the selective enrichment of target proteins on this grid. Using GFP11-tagged apoferritin, we demonstrate fluorescence-guided protein capture and obtain a 2.58 Å cryo-EM reconstruction, establishing SGFP-G as an affinity-grid platform for high-resolution structural studies with reduced sample requirements.
Language: English
Subfield: Structural Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7212739154