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Automated Generation of Supported Lipid Bilayer Arrays with Controlled Receptor Densities in Well Plates
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DOI：10.1021/acsami.6c02551.png)
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Abstract
En 中文
Understanding the superselective, multivalent interactions that drive immunity, infection, and biosensing requires (i) model surfaces with precisely tunable receptor density and (ii) quantitative readouts. Here, we introduce a fully automated well-plate-based platform using cell-mimicking supported lipid bilayers (SLBs) that enables both requirements with standardized, commercially available equipment. We outline the main challenges associated with integrating liquid handling with the shear and disruption-sensitive SLB system and offer practical guidelines to overcome them. The resulting versatile and scalable workflow yielded high-quality antifouling surfaces without laborious manual pipetting, while preserving the simplicity of pipet and well-plate-based liquid handling. It enables complex assays requiring more than 1000 aspiration and dispensing steps over >12 h while maintaining surface integrity. Control over the receptor density was achieved by variation of the molar fraction of a biotin-functionalized lipid inside the SLB, to which streptavidin and biotinylated receptors were bound using the strong biotin–streptavidin interaction. A quantitative readout of the receptor density employing fluorescently labeled streptavidin was statistically validated in the low pmol·cm–2 range. The generated arrays were applied in two biorecognition studies requiring elaborate liquid-handling procedures. DNA hybridization showcases a strong, specific, and stoichiometric binding of fluorescently labeled complementary DNA. Furthermore, the platform was established as an alternative type of glycan array capable of studying the complex, multivalent binding of labeled virus particles scalable to the high-throughput processing of samples.
Keywords:
Fluorescence
Genetics
Lipids
Receptors
Vesicles
supported lipid bilayers
receptor density control
fluorescence quantification
multivalent interactions
glycan arrays
DNA hybridization
biosensing
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