Thursday, 3 November 2005 - 12:50 PM
526b
Simple Bioconjugation and Separation of Luminescent Cdse-Zns Quantum Dots by Genetically Engineered Polyhistidine-Tagged Elp-Protein L Fusions as a Sensitive Reagent for Immuoassay
U. Loi Lao1, Gabriela Jenikova
1, Ashok Mulchandani
2, and Wildred Chen
3. (1) Chemical and Environmental Engineering, University of California, 900 University Avenue, Riverside, CA 92521, (2) Department of Chemical and Environmental Engineering, University of California, Riverside, CA 92521, (3) University of California, 900 University Avenue, Riverside, CA 92521
A simple platform for the direct bioconjugation and separation of highly luminescent CdSe-ZnS quantum dots (QD) was reported through metal binding coordination with genetically engineered N-terminal polyhistidine tagged elastin (ELP)-Protein L fusion. The principle of immunoassay-ready conjugates was to take advantage of the unique temperature-responsive property of ELP in combination with the high affinity of the antibody-binding Protein L domain. Simple separation of the QD-ELP-ProL complex was achieved by thermally-triggered precipitation and no interference on QD functionality was observed after several reversible transition cycles. A variety of antibodies were shown to bind to Protein L with high affinity, allowing us to perform different formats of immunoassay with versatility. We believed this simple conjugation-separation of QD with genetically engineered protein will facilitate the analysis for diverse analytes.
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