Assembly of bionanostructures onto beta-cyclodextrin molecular printboards for antibody recognition and lymphocyte cell counting

Manon J W Ludden, Xiao Li, Jan Greve, Aart van Amerongen, Maryana Escalante, Vinod Subramaniam, David N Reinhoudt, Jurriaan Huskens

    Research output: Contribution to JournalArticleAcademicpeer-review


    The assembly of complex bionanostructures onto beta-cyclodextrin (betaCD) monolayers has been investigated with the aims of antibody recognition and cell adhesion. The formation of these assemblies relies on host-guest, protein-ligand, and protein-protein interactions. The buildup of a structure consisting of a divalent bis(adamantyl)-biotin linker, streptavidin (SAv), biotinylated protein A (bt-PA), and an Fc fragment of a human immunoglobin G (IgG-Fc) was studied with surface plasmon resonance (SPR) spectroscopy. Patterns of this bionanostructure were obtained via microcontact printing of the divalent linker at the molecular printboard, followed by the subsequent attachment of the proteins. Fluorescence microscopy showed that the buildup of these bionanostructures on the betaCD monolayers is highly specific. On the basis of these results, bionanostructures were made in which whole antibodies (ABs) were used instead of the IgG-Fc. These ABs were bound to the SAv layer via biotinylated protein G (bt-PG) or via a biotinylated AB. These constructions yielded specifically bound ABs with a less than maximal density, as shown by SPR spectroscopy and atomic force microscopy (AFM). Finally, the immobilization of ABs to the molecular printboard was used to create platforms for lymphocyte cell count purposes. Monoclonal ABs (MABs) were attached to the SAv layer using bt-PG, an engineered biotin functionality, or through nonspecific adsorption. The binding specificity of the immobilized cells was the highest on the buildup made from bt-PG, which is attributed to an optimized orientation of the antibodies. An approximately linear relationship between the numbers of seeded cells and counted cells was demonstrated, rendering the platform potentially suitable for lymphocyte cell counting.

    Original languageEnglish
    Pages (from-to)6964-73
    Number of pages10
    JournalJournal of the American Chemical Society
    Issue number22
    Publication statusPublished - 4 Jun 2008


    • Biotin
    • Humans
    • Immunoglobulin Fc Fragments
    • Immunoglobulin G
    • Lymphocyte Count
    • Microscopy, Atomic Force
    • Nanostructures
    • Spectrometry, Fluorescence
    • Streptavidin
    • Surface Plasmon Resonance
    • beta-Cyclodextrins
    • Journal Article
    • Research Support, Non-U.S. Gov't


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