Research Project
6736476
DESCRIPTION (provided by applicant): This proposal seeks support for the commercial development of a new class of fluorescent lanthanide complexes of dramatically enhanced brightness that will be used in protein assays and protein on a chip assay development in an homogeneous and multiplex format. Protein microarrays are an attractive technology for rapid screening of large numbers of proteins in small amounts, similar to DNA microarrays. However, sensitivity is a special challenge in protein analysis because, unlike DNA analysis, there are no available options for amplification of protein levels. The only way to measure low levels of proteins is through more sensitive detection methods. Organic-fluorescence-based methods currently offer the best options, but better ones are needed. Lumiphore's lanthanide complexes offer unprecedented sensitivity that will enable protein assays and related technology. The long fluorescence lifetime of lanthanide complexes enable time resolved fluorescence assays (thereby lowering substantially the background fluorescence from assay solutions). The fundamental coordination chemistry of these compounds has been studied and developed at Berkeley and two broad UC patents have resulted and are exclusively licensed to Lumiphore. These lanthanide complexes of isophthalamide and salicylamide ligands are stable in aqueous solution to very low concentrations and capture and emit light much more efficiently than existing commercial agents. Enhancement of assay sensitivity of up to a factor of 100 or more is enabled by the high quantum yield and additional brightness of these new agents. To develop new partner assays using our technology, Lumiphore has already established a number of industry partnerships. In this application Lumiphore proposes to develop new assays using its proprietary technology for attachment of the luminescent lanthanide complex to proteins and the development of protein chips. In the phase I project specific binding of the lanthanide complex on 96/384 well plates will be demonstrated and the attachment of the lanthanide complex to proteins/antibodies will be optimized using several chemical linkers. The second phase will consist of the development and optimization of the specific binding of a lanthanide complex to glass slides to produce protein chips. Protein chips using antibodies constitute an emerging technology that addresses the need for increased throughput and decreased sample size for analysis of protein concentration, expression levels, structure-function relations and protein-protein interaction. Existing fluorescence detection technology is useful but suffers from a lack of sensitivity and multiplexing. Other methods, such as those using gold and silver particles and chemiluminescence, are useful but have limitations such as limited sensitivity, heterogeneous assays and no multiplexing. The availability of quantum yields of over 60% and multiplex assays based on simultaneous use of more than one lanthanide complex can enable entire new types of assays, substantially enhancing the sensitivity of existing assays, and will ultimately enable the development of new protein assays on a chip. Key words: time resolved fluorescence, lanthanide fluorescence, protein assays on a chip, high throughput assays, multiplex and homogenous assays.
