THROUGH-BOND ENERGY TRANSFER CASSETTES FOR MULTIPLEXING & DEVELOPMENT OF METHODS FOR PROTEIN MONO-LABELING
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Date
2010-07-14
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Abstract
A set of three through-bond energy transfer cassettes based on BODIPY as a donor
and cyanine dyes as acceptors has been prepared via Sonogashira couplings, and their
photophysical properties were examined. These cassettes fluoresce around 600 to 800
nm and are resolved by approximately 100 nm. This property is an important factor for
multiplexing study in cellular imaging. Several useful fluorescent probes such as 5- and
6-carboxyfluorescein, water-soluble BODIPY, and water-soluble Nile Blue dyes, have
also been synthesized and their photophysical properties studied.
We have also attempted to develop a method for protein mono-labeling via a solidphase
approach. The labeling of protein with one fluorescent dye facilitates
quantification and single molecule imaging in biological applications. Various solidsupports
such as PEGA, CPG, and BSA-coated CPG, were tested. Photolabile and
chemically cleavable linkers were prepared to connect solid-supports and fluorophores.
Unfortunately, our approach to the fluorescent mono-labeling of native proteins did not
give us any conclusive results.
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Keywords
through-bond energy transfer, fluorescent dye, water-solubility, protein mono-labeling, solid-phase