Hydrogels for biomolecule analysis and corresponding method to analyze biomolecules
US7588906B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Dec 16, 2005 |
| Grant date | Sep 15, 2009 |
| Priority date | — |
| Expiry date | Jan 1, 2027 |
Classification
- Technology area (CPC H)Electricity
- CPC primaryH01J49/04
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Polyacrylamide-based methods of fabricating surface-bound peptide and protein arrays, the arrays themselves, and a method of using the arrays to detect biomolecules and to measure their concentration, binding affinity, and kinetics are described. Peptides, proteins, fusion proteins, protein complexes, nucleic acids, and the like, are labeled with an acrylic moiety and attached to acrylic-functionalized glass surfaces through a copolymerization with acrylic monomer. The specific attachment of glutathione S-transferase-green fluorescent protein (GST-GFP) fusion protein was more than 7-fold greater than the nonspecific attachment of non-acrylic labeled GST-GFP. Surface-attached GST-GFP (0.32 ng/mm2) was detectable by direct measurement of green fluorescent protein fluorescence and this lower detection limit was reduced to 0.080 ng/mm2 using indirect antibody-based detection. The polyacrylamide-based surface attachment strategy was also used to measure the kinetics of substrate phosphorylation by the kinase c-Src which is encoded by the Rous Sarcoma virus. The surface attachment strategy is applicable to the proteomics field and addresses denaturation and dehydration problems associate…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.