A new compact disc format of high density array synthesis applied to peptide nucleic acids and in situ MALDI analysis

Abstract

A fully automated synthesizer was constructed and designed to perform high speed miniaturized syntheses of compound libraries using the SPOT technique. Utilizing magnetically controlled drop-on-demand ink jet nozzles, an r/phi array format of 2500 spots can be simultaneously dispensed from up to 24 separate reagent valves onto a rotating disc as the solid phase in less than three minutes. In addition, a complete wash station is on board allowing for fully programmable combinatorial syntheses without manual attention. A new carbon black/polypropylene composite solid phase disc was developed and tested for its functionalisation/loading, spot detection, durability and MALDI-TOF target capabilities. The carbon black/polypropylene composite was then successfully employed jointly as the solid phase in the syntheses of short peptide and PNA oligomers and as the target probe holder for MALDI-TOF measurement without transfer of the material. Several protocols for PNA syntheses were also investigated and an optimised PNA methodology for the carbon black/polypropylene composite is reported.

Miniaturized and Automated Synthesis of Biomolecules—Overview and Perspectives

“…Due to the cellulose moieties, these peptides were easily immobilized onto a glass surface by physical absorption 141a,154. Alternatively, biological interactions (biotin/streptavidin or avidin/biotin) or covalent linkage (amine/aldehyde, maleimide/thiol, or amine/carboxy chemistry) were used to tether these peptides to a surface and in array format.…”

Peptide Arrays: Development and Application

“…21 Similary, Dikmans and colleagues grafted amino poly(ethylene glycol) (PEG) functionality into a polypropylene support which allowed subsequent coupling of peptides onto the support. 52…”

Microarray Technology as a Universal Tool for High-Throughput Analysis of Biological Systems

“…In this case the peptides remain bound to the dissolved cellulose molecules, which build a three dimensional structure on the re-arrayed spots of the resulting glass-chips. On the other hand, a Bio DiscSynthesizer was developed, aiming at a fully automated SPOT synthesis at high speed [140]. Using a drop-ondemand technology, this machine is capable of simultaneously spotting 2500 spots from up to 24 separate reagent valves onto a rotating disc as the planar array substrate in less than three minutes.…”