title: Conjugation of biotin-coated luminescent quantum dots with single domain antibody-rhizavidin fusions.
abstract: Straightforward and effective methods are required for the bioconjugation of proteins to surfaces and particles. Previously we demonstrated that the fusion of a single domain antibody with the biotin binding molecule rhizavidin provided a facile method to coat biotin-modified surfaces with a highly active and oriented antibody. Here, we constructed similar single domain antibody-rhizavidin fusions as well as unfused rhizavidin with a His-tag. The unfused rhizavidin produced efficiently and its utility for assay development was demonstrated in surface plasmon resonance experiments. The single domain antibody-rhizavidin fusions were utilized to coat quantum dots that had been prepared with surface biotins. Preparation of antibody coated quantum dots by this means was found to be both easy and effective. The prepared single domain antibody-quantum dot reagent was characterized by surface plasmon resonance and applied to toxin detection in a fluoroimmunoassay sensing format.
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: Jinny L
authors.initials: JL
authors.lastname: Liu
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Walper
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: Kendrick B
authors.initials: KB
authors.lastname: Turner
authors.affiliation: NOVA Research Inc., 1900 Elkin St Suite 230, Alexandria, VA 22308, USA.
authors.forename: Audrey Brozozog
authors.initials: AB
authors.lastname: Lee
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: Igor L
authors.initials: IL
authors.lastname: Medintz
authors.affiliation: Sotera Defense Solutions, Inc., 7230 Lee Deforest Drive, Columbia, MD 21046, USA.
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Susumu
authors.affiliation: Sotera Defense Solutions, Inc., 7230 Lee Deforest Drive, Columbia, MD 21046, USA.
authors.forename: Eunkeu
authors.initials: E
authors.lastname: Oh
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Zabetakis
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: Ellen R
authors.initials: ER
authors.lastname: Goldman
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA.
authors.forename: George P
authors.initials: GP
authors.lastname: Anderson
keywords: Quantum dots; Rhizavidin; Single domain antibodies; Surface plasmon resonance