Chemical array technology and screening, preparation of Chemical beads

   Contact: Kaori Honda  E-mail: tech_CRAST
   Collaborative Scientist: Makoto Kawatani

Chemical Array

 Chemical arrays is one of the approaches of high-throughput screening utilize physical interaction. We have developed a unique photo-cross-linking technique for immobilizing variety of chemicals in a functional group independent manner. This technique enables the immobilization of 3,456 natural and synthetic compounds on single solid surface without any functional group derivatization. We manufactured 12 chemical array series (NPDepoArray) with 30,000 NPDepo-Library compounds in total (as of 2022.03.24).
 Purified sample with tags (e.g. His, GST, FLAG) are preferable for array screening. After incubating with sample, arrays are incubated with 1st antibody for tags, then Cy5 labeled 2nd antibody. The fluorescence signal of arrays are then scanned with array scanner.

Chemical Beads

 The molecular target of the chemical which induce a specific phenotype in cellular system were identified by pull down assay. However, it remains challenging and time-consuming to derivatize chemicals into tagged probes for pull-down assay without a loss of biological activity.
 We developed a chemical beads which can immobilize chemicals on to beads surface in a functional group-independent manner. That is, commercially available NHS activated sepharose beads were coupled with photo-affinity linker and chemicals were immobilized by nonselective photo-cross-linking method. This method implement an exposure of variety functional groups of chemical. This could be used to identify and purify the binding proteins from cell lysate.

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