Lipoconjugates for the noncovalent generation of microarrays in biochemical and cellular assays.
The generation of microarrays by functionalization of hydrophobic glass surfaces with conjugates of triacylated lipophilic end-groups and with a peptide or hapten as a test substance is presented. Immobilization on the hydrophobic surfaces through the triacylated anchor group is fully orthogonal to the reactivity of functional groups within the test substances. The technique is therefore free of risk that reactions of these functional groups may influence the biological activity of the test compounds in screening applications. In addition, no preactivation of either the surface or the compounds is required. Reagents and substrates may be stored at ambient conditions for long periods of time. The lipoconjugates are administered from aqueous solution enabling automated nanopipetting down to spot dimensions of 100 microm across. The microstructures are stable with respect to the conditions of biochemical assays and applications in cell biology. Due to the hydrophobicity of the nonfunctionalized surfaces, standard blocking protocols used in microtiter-plate testing can be employed, thereby inhibiting nonspecific binding of assay reagents. Generation of these microstructures on hydrophobic glass slides or coverslips enables highly sensitive multichannel read-outs with high-resolution fluorescence microscopy.
Hoff
Antje
A
Group of Genomics and Proteomics, Center for Bioinformatics Tübingen, Institute for Cell Biology, University of Tübingen, Auf der Morgenstelle 15, 72076 Tübingen, Germany.
André
Thomas
T
Schäffer
Tilman E
TE
Jung
Günther
G
Wiesmüller
Karl-Heinz
KH
Brock
Roland
R
D000954:Antigens, Surface / Q000378:metabolism*; D019921:Biotinylation; D020013:Calcium Signaling; D020650:Combinatorial Chemistry Techniques; D003584:Cytological Techniques; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D019169:Jurkat Cells; D008055:Lipids; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*
D000954:Antigens, Surface; D008055:Lipids; D011506:Proteins