Expedited High-Affinity Ligand Generation from Biomarker Sequence by Integration of Data Mining with High-throughput Screening

Employing the fields of toxico-genomics, proteomics, and computer modeling has helped identify novel biomarkers that indicate early preclinical damage to various organ systems, offering promise of their wide applicability for monitoring and pre-emptive preservation of military and civilian health. The realization of this promise impinges on the availability of sensitive and selective affinity reagents for their detection. Existing methodologies for generation of affinity reagents are cost prohibitive and labor intensive. A niche exists for obtaining high affinity ligands to newly identified biomarker proteins merely from their primary sequence information. In Phase I we showed proof-of-concept for a high affinity ligand generation system. The process combines an innovative datamining methodology with an automated high-throughput screening method to produce high affinity ligands to protein biomarkers. Information in existing proteomics databases was exploited (using our automated mining software to be implemented in Phase II) to find high affinity ligand candidates. A custom high-density peptide microarray of these candidates was created and screened using the protein to identify high-affinity ligands experimentally. Using this methodology, it will be possible to produce very high affinity recognition agents for biomarkers very rapidly, in a matter of days, easily besting all other existing techniques.