Fluorophore-Conjugated Holliday Junctions for Generating Super-Bright Antibodies and Antibody Fragments
Author(s)Li, Zeyang; Theile, Christopher S.; Chen, Guan-Yu; Bilate, Angelina M.; Duarte, Joao N.; Avalos, Ana M.; Fang, Tao; Barberena, Roberto; Sato, Shuji; Ploegh, Hidde; ... Show more Show less
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The site-specific modification of proteins with fluorophores can render a protein fluorescent without compromising its function. To avoid self-quenching from multiple fluorophores installed in close proximity, we used Holliday junctions to label proteins site-specifically. Holliday junctions enable modification with multiple fluorophores at reasonably precise spacing. We designed a Holliday junction with three of its four arms modified with a fluorophore of choice and the remaining arm equipped with a dibenzocyclooctyne substituent to render it reactive with an azide-modified fluorescent single-domain antibody fragment or an intact immunoglobulin produced in a sortase-catalyzed reaction. These fluorescent Holliday junctions improve fluorescence yields for both single-domain and full-sized antibodies without deleterious effects on antigen binding.
DepartmentMassachusetts Institute of Technology. Department of Biology; Whitehead Institute for Biomedical Research
Angewandte Chemie International Edition
Li, Zeyang et al. “Fluorophore-Conjugated Holliday Junctions for Generating Super-Bright Antibodies and Antibody Fragments.” Angewandte Chemie International Edition 54.40 (2015): 11706–11710.
Author's final manuscript