A photophysical study of two fluorogen-activating proteins bound to their cognate fluorogens

Tiziano Gaiotto; Hau B. Nguyen; Jaemyeong Jung; Gnana S. Gnanakaran; Jurgen G. Schmidt; Geoffrey S. Waldo; Andrew M. Bradbury; Peter M. Goodwin

doi:10.1117/12.875418

11 February 2011 A photophysical study of two fluorogen-activating proteins bound to their cognate fluorogens

Tiziano Gaiotto, Hau B. Nguyen, Jaemyeong Jung, Gnana S. Gnanakaran, Jurgen G. Schmidt, Geoffrey S. Waldo, Andrew M. Bradbury, Peter M. Goodwin

Author Affiliations +

Proceedings Volume 7905, Single Molecule Spectroscopy and Imaging IV; 79050O (2011) https://doi.org/10.1117/12.875418
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

We are exploring the use of fluorogen-activating proteins (FAPs) as reporters for single-molecule imaging. FAPs are single-chain antibodies selected to specifically bind small chromophoric molecules termed fluorogens. Upon binding to its cognate FAP the fluorescence quantum yield of the fluorogen increases giving rise to a fluorescent complex. Based on the seminal work of Szent-Gyorgyi et al. (Nature Biotechnology, Volume 26, Number 2, pp 235-240, 2008) we have chosen to study two fluorogen-activating single-chain antibodies, HL1.0.1-TO1 and H6-MG, bound to their cognate fluorogens, thiazole orange and malachite green derivatives, respectively. Here we use fluorescence correlation spectroscopy to study the photophysics of these fluorescent complexes.

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Tiziano Gaiotto, Hau B. Nguyen, Jaemyeong Jung, Gnana S. Gnanakaran, Jurgen G. Schmidt, Geoffrey S. Waldo, Andrew M. Bradbury, and Peter M. Goodwin "A photophysical study of two fluorogen-activating proteins bound to their cognate fluorogens", Proc. SPIE 7905, Single Molecule Spectroscopy and Imaging IV, 79050O (11 February 2011); https://doi.org/10.1117/12.875418

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