High-Speed Label-Free Optical Detection System for Small-Molecule Microarrays

  • Zhu, Xiangdong, (PI)

Project: Research project

Description

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop a robust, sensitive and reliable label-free detector that can evaluate the real-time binding kinetics of 10,000 spot small molecule chemical microarrays against various single component or multi-component analytes. In 1991, we first reported the one-bead one-compound combinatorial library (OBOC) method which is highly efficient and we have successfully used it to identify ligands to a large number of biological targets. Although the OBOC library method is highly versatile, the amount of compounds contained in one single bead is small (100 pico-mol) and is inadequate for many solution phase assays. Last year, we reported on the development of an encoded "one-aggregate one-compound" library method, in which the powerful split-synthesis method is used for the construction of the library, and compounds at l -10 mmol range can be efficiently prepared. Very recently, we reported on the development and application of a novel chemical microarray method, in which small organic molecules or short peptides are chemo-selectively ligated to a polymer (e.g. agarose or protein) and then printed on glass, plastic microscope slides or PVDF membranes. We envision that by combining our highly efficient "one-aggregate one-compound" library method with this novel chemical microarray technique, we easily can print thousands of replicates of high density small molecule microarrays (10,000 spots/ slide) and use them to probe a variety of biological analytes such as serum, cell extracts or pure proteins. Our hypothesis is that by combining our novel combinatorial chemistry and microarray platforms with a highly sensitive and reliable 2-D label-free optical detector that can efficiently measure real-time binding kinetics, we will be able to rapidly and accurately study the binding kinetics of a large number of analytes (e.g., individual protein or complex analytes such as whole serum) against a large number of immobilized small molecules, peptides, oligonucleotides or proteins. Specific aims of this proposal include the design and construction of a prototype detector that can measure real-time binding kinetics of analytes (dissolved in the mobile phase) against 100 - and 400-spot chemical microarrays. The next phase will be to develop a detector that can analyze 10,000 spot microarrays in one single run. Five 10,000 small molecule encoded bead-aggregate libraries will be prepared and printed on glass slide replicates as microarrays, and these microarrays will be analyzed by the optical detector to be developed in this proposed research.
StatusFinished
Effective start/end date1/17/0611/30/10

Funding

  • National Institutes of Health: $443,841.00
  • National Institutes of Health: $151,000.00
  • National Institutes of Health: $587,215.00
  • National Institutes of Health: $548,379.00
  • National Institutes of Health: $517,898.00

Fingerprint

Microarrays
Labels
Molecules
Detectors
Kinetics
Glass
Peptides
Proteins
Oligonucleotides
Sepharose
Assays
Polymers
Microscopes
Plastics
Ligands
Membranes

Keywords

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)