Small Molecule Microarray Protocol

Fluorous modified glass slides have been used by several groups for the immobilization of fluorous tagged molecules for microarray formation.  Drs. Angela Koehler and Arturo Vegas at the Broad Institute of MIT and Harvard were amongst the first groups to take advantage of this novel immobilization method for the formation of small molecule microarrays.  Their work was published in Angewandte Chemie Intl. Ed. a few years back and we commented on the work in a F-Blog post.  In that work they interrogated potential small molecule inhibitors of various histone deactylases using fluorous microarrays.  The putative inhibitors which were identified were then confirmed by SPR and microtitre plate based assays, thus validating the fluorous microarray format.  The fluorous format produced better spot morphology, increased signal to noise, and simpler workflows.  In addition the low non-specific binding observed with the fluorous surface allowed interrogation either with isolated enzymes or with cell lysates.

The work was particularly interesting to us at FTI because it verified that a single fluorous tag can immobilize a very large complex of molecules under the proper conditions.  In this instance a small molecule-protein-antibody complex remained in place.  Remarkable when you think about it considering there is no covalent bond, no affinity, no charge interaction taking place.  Just partitioning.

As a follow-up to that paper, the two researchers have just published a detailed description and protocol of fluorous small molecule microarray formation as a chapter in the book “Small Molecule Microarrays” that is part of the Methods in Molecular Biology series published by Springer.  Included within the protocol are notes which highlight potential trouble spots and solutions.  For those interested in probing protein-small molecule interactions using fluorous microarrays this detailed protocol is highly recommended.

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