Ribosome display and dip-pen nanolithography for the fabrication of protein nanoarrays

A combination of the ribosome display technique and DPN used to fabricate protein nanoarrays without prior protein purification was investigated. The investigation used a cell-free system for the production of protein, which is particularly useful for the rapid production of proteins based only on genome information. The target EGFP and linker λ-DNA were amplified through PCR. The full size target DNA for ribosome display was also prepared by assembly PCR and used as a template for in vitro transcription. Glass substrates were cleaned with 'piranha' solution for 60 min at 69°C in a sonic cleaner. It was observed that the fusion of molecules permit selective hybridization between oligonucleotides on the nanoarray surface and the mRNA portion of fusion molecules and can convert the oligonucleotides to a protein nanoarray.