Work Package 3: SPRi Detection of Peptide-Receptor Interactions – University of Copenhagen

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Work Package 3: Label-free IMS of peptide microarrays

  • Develop label-free MALDI-IMS methods for analysis of peptides and proteins in high-density pep-tide microarrays.
  • Increase the analysis speed of MALDI-IMS of high-density arrays by focusing on selected analysis points obtained by pre-screening of the arrays in SPRi.

  • Increase the analysis speed by development of prototype instrumentation with “smart beam” laser technology. 

  • Develop software platforms for analysis of MS-data from peptide arrays.

P8 has obtained proof-of-concept for the label-free detection of peptides attached to gold surfaces by MALDI-TOF MS. P3 will in collaboration with P7 and P8 refine the methodology to make it suitable for analysis of the small peptide fields in the arrays made by P2 Different matrices, linkers between peptides and the solid phase used in synthesis will be evaluated using the instrumentation developed by P7.

The sensitivity, reproducibility and compatibility with SPR imaging will be validated in order to define a fast and efficient MALDI-IMS of peptide chip arrays. P7 will design adapters for the chips used in the peptide arrays and will in collaboration with P2 and P3 integrate recording of peptide arrays into the imaging software for the MALDI-IMS device.

Partner responsibilities (Instrument development):

  • P7 will develop a MALDI-IMS instrument prototype with enhanced imaging speed.

  • P3 and P7 will collaborate with P4 on development of software that will use coordinates from target regions selected from SPRi of peptide arrays as input to MALDI devices that can then confine their analysis to the selected coordinates.
  • P2, P3 and P7 will collaborate on development of software for identification of proteins binding to peptides in high-density peptide arrays analyzed by MALDI-IMS.

  • P8 is responsible for developing MALDI desorption of peptide microarrays
  • P7 for developing MALDI instruments
  • P3 for performing MALDI IMS analysis of peptide microarrays

Partner responsibilities (substrate and application development):

  • All partners will be responsible for designing and/or analyzing peptide microarrays using MALDI IMS.
  • P5 for developing surface coatings
  • P6 for developing 3D surfaces
  • P4 for integrating SPRi detection and MALDI IMS for peptide microarray analysis
  • P10 for developing bioinformatics tools to analyze MALDI
IMS detection of peptide microarrays
  • P1 for analyzing immune reactions using MALDI IMS detection of microarrays.