HIPAD

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High-Density Peptide MicroArrays and high-throughput, label-free
detection of peptides, modifications and interactions” is a FP7 project

Project number 278832

With more than 6 million budget over 4 years.

Site: http://hipad.ku.dk/theproject/

Summary of the Project

Proteins are extremely malleable building blocks of life involved in all aspects of biology. Many diseases are caused by proteins aberrations, and proteins are frequent targets of intervention.

Mapping all proteins and
their functions are expected to yield pervasive medical and biotechnological benefits.

However, even the most comprehensive and high-throughput proteins discovery technologies are seriously challenged by the extreme diversity and low abundance of many proteome components; a problem, compounded by the lack of affinity reagents and validated probes for sample preparation and identification.

Our concept is that shorter protein fragments, or peptides, may offer solutions to many of these problems as peptides may represent or mimic proteins. Using in situ solid-phase peptide synthesis, computerized photolithography and novel photochemistry, we have recently generated peptide microarrays of up to 2 million addressable peptides.

This unprecedented high-density and high-content peptide microarray technology could make inroads into the kind of high-throughput analysis needed to address the entire human proteome. Here, we aim to exploit this potential by using and improving three different, yet complementary, label-free detection technologies allowing sensitive, high-resolution determinations of the identity, quality and/or modification of individual members of a peptide microarray, and real-time monitoring of any interacting molecular receptor.

We will also develop peptides as rapid, specific, and renewable affinity reagents for complex sample preparation, and develop peptides as probes and complex biosensors. Three SME’s constitute the backbone of this collaboration, receiving 50% of the budget, and enjoying significant opportunities from the booming protein/peptide microarray market. Furthermore, solutions to these unmet needs of proteomics are believed to have incalculable benefits for European health, innovativeness and competiveness.

The main goal of Hi-PAD is to develop a High Density Peptide Array reader based on PLasmore technology. The reader, called HI-TEC, will exploit the main advantages of nanoplasmonic surfaces, which include an extreme simplification of the optical scheme and a striking improvement of the local sensitivity to biomolecules.

Thus, it is possible to decrease the size/field and increase the detector area leading to a considerable increase in the resolution compared to current SPRi technology, and a reduction in costs. The HI-TEC will allow parallel real-time monitoring of protein interaction with hundreds of thousands of peptides at a time and can lead to rapid identification of peptide ligands to protein receptors, antibodies and other macromolecular peptide receptors.

The HI-TEC is not conceived to be a stand-alone instrument. Within the HI-PAD project but also for concrete marketing reasons, the system will be a very powerful accessory for MALDI (Matrix Assisted Laser Desorption Ionization) imaging systems.
An outstanding advancement in the field of proteomics will be given by coupling or integrating the HI-TEC with MALDI imaging technology. The HI-TEC will serve as a pre-screening tool for the fast individuation of the spots of the HDA which have immobilized the ligands.

Once screened by the HI-TEC the chip will be transferred in a MALDI imaging system for the protein characterization. The pre-screening will permit a much faster readout of the HDA by the MALDI imaging since only the spots which have immobilized the ligands will be analyzed. Plasmore will then use the great advantage of knowing the existing market of MALDI users.

Plasmore has also the option of a joint-venture with MALDI producers (for example, one of the MALDI leaders of the market, Bruker Daltonics http://www.bdal.com/ is a member of the project HI-PAD). Together with the analytical device Plasmore will develop the relative sensing chip (HI-Chip), a plasmonic chip based on Plasmore technology which can be used by iSPR with the HI-TEC and also with MALDI imaging systems.