Your browser doesn't support javascript.
loading
Design, Synthesis, Conjugation, and Reactivity of Novel trans,trans-1,5-Cyclooctadiene-Derived Bioorthogonal Linkers.
Longo, Beatrice; Zanato, Chiara; Piras, Monica; Dall'Angelo, Sergio; Windhorst, Albert D; Vugts, Danielle J; Baldassarre, Massimiliano; Zanda, Matteo.
  • Longo B; Institute of Medical Sciences, Foresterhill, University of Aberdeen, AB252ZD Aberdeen, United Kingdom.
  • Zanato C; Centre for Sensing and Imaging Science, School of Science, Loughborough University, LB11 3TU Loughborough, United Kingdom.
  • Piras M; Institute of Medical Sciences, Foresterhill, University of Aberdeen, AB252ZD Aberdeen, United Kingdom.
  • Dall'Angelo S; Institute of Medical Sciences, Foresterhill, University of Aberdeen, AB252ZD Aberdeen, United Kingdom.
  • Windhorst AD; Institute of Medical Sciences, Foresterhill, University of Aberdeen, AB252ZD Aberdeen, United Kingdom.
  • Vugts DJ; Amsterdam UMC, Vrije Universiteit, dept. Radiology and Nuclear Medicine, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
  • Baldassarre M; Amsterdam UMC, Vrije Universiteit, dept. Radiology and Nuclear Medicine, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
  • Zanda M; Institute of Medical Sciences, Foresterhill, University of Aberdeen, AB252ZD Aberdeen, United Kingdom.
Bioconjug Chem ; 31(9): 2201-2210, 2020 09 16.
Article en En | MEDLINE | ID: mdl-32786505
ABSTRACT
The tetrazine/trans-cyclooctene (TCO) inverse electron-demand Diels-Alder (IEDDA) reaction is the fastest bioorthogonal "click" ligation process reported to date. In this context, TCO reagents have found widespread applications; however, their availability and structural diversity is still somewhat limited due to challenges connected with their synthesis and structural modification. To address this issue, we developed a novel strategy for the conjugation of TCO derivatives to a biomolecule, which allows for the creation of greater structural diversity from a single precursor molecule, i.e., trans,trans-1,5-cyclooctadiene [(E,E)-COD] 1, whose preparation requires standard laboratory equipment and readily available reagents. This two-step strategy relies on the use of new bifunctional TCO linkers (5a-11a) for IEDDA reactions, which can be synthesized via 1,3-dipolar cycloaddition of (E,E)-COD 1 with different azido spacers (5-11) carrying an electrophilic function (NHS-ester, N-succinimidyl carbonate, p-nitrophenyl-carbonate, maleimide) in the ω-position. Following bioconjugation of these electrophilic linkers to the nucleophilic residue (cysteine or lysine) of a protein (step 1), the resulting TCO-decorated constructs can be subjected to a IEDDA reaction with tetrazines functionalized with fluorescent or near-infrared (NIR) tags (step 2). We successfully used this strategy to label bovine serum albumin with the TCO linker 8a and subsequently reacted it in a cell lysate with the fluorescein-isothiocyanate (FITC)-derived tetrazine 12. The same strategy was then used to label the bacterial wall of Gram-positive Staphylococcus aureus, showing the potential of these linkers for live-cell imaging. Finally, we determined the impact of structural differences of the linkers upon the stability of the bioorthogonal constructs. The compounds for stability studies were prepared by conjugation of TCO linkers 6a, 8a, and 10a to mAbs, such as Rituximab and Obinutuzumab, and subsequent labeling with a reactive Cy3-functionalized tetrazine.
Asunto(s)

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Alcadienos / Colorantes Fluorescentes Límite: Animals Idioma: En Año: 2020 Tipo del documento: Article

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Alcadienos / Colorantes Fluorescentes Límite: Animals Idioma: En Año: 2020 Tipo del documento: Article