Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

The Fastest and the Brightest

07.06.2013
BODIPY–tetrazine derivatives as superbright bioorthogonal turn-on probes

American researchers have developed a probe for marking biomolecules that begins to fluoresce only when it is “switched on” by binding. As reported in the journal Angewandte Chemie, the reaction takes place very quickly and the difference in brightness between the “on” and “off” states is two orders of magnitude bigger than for conventional activatable probes.

Marking biomolecules in living cells with fluorescent probes is a well-established technique. New research possibilities open up when these probes are combined with bioorthogonal reactions. Such reactions can occur inside a living system without disrupting normal biochemical processes.

This makes it possible to generate “turn-on” probes: a bioorthogonal reaction binding partner is bound to the biomolecule of interest (without affecting it) and acts as an anchoring site for the fluorescent probe. The probe is devised so that its fluorescence is significantly increased when it binds to the anchoring site. Because the probes not bound to the target fluoresce far less, background fluorescence is reduced. This eliminates the need for complex washing procedures that delay observation of the cells.

For all of this to work, the probe system must work without a toxic catalyst, react quickly to allow for time-resolved observation of biological processes, and fluoresce very strongly after being “turned on” to maximize the signal–strength relative to the background. It has not previously been possible to meet all of these requirements in one system.

A team led by Ralph Weissleder at Massachusetts General Hospital and Harvard University has now developed a system that fits the bill: an unusually bright, fast reacting, biocompatible probe system with a large difference between the switched on and switched off states.

The new probe consists of two components: The first is a fluorescent dye called BODIPY (boron dipyrromethene), a three-ring system with a subunit made of one boron, two nitrogen, and two fluorine atoms. The second component is a tetrazine molecule, a six-membered ring containing four nitrogen and two carbon atoms. Tetrazine quenches the fluorescence of BODIPY, which passes incoming energy off to the tetrazine component without radiation instead of fluorescing.

Tetrazine simultaneously serves as a reagent for the bioorthogonal reaction.
The reaction partner is modified trans-cyclooctene (TCO), which the researchers couple to the biomolecule to be studied by means of an antibody. When the probe is added, the tetrazine binds to the TCO, giving off nitrogen and binding the probe to the biomolecule.

The reaction destroys the probe’s tetrazine group, turning off the quenching of the fluorescence and allowing the BODIPY molecule to glow an intense green. The researchers recorded fluorescence over a thousand times stronger than that of the probe in the “off” state. This is two orders of magnitude stronger than all previously described turn-on probes.

The success of this system is due to the particularly strong fluorescence quenching made possible by the special electronic constellation and spatial arrangement of the BODIPY and tetrazine components relative to each other.

About the Author
Dr. Weissleder is Professor at Harvard University, Director of the Center for Systems Biology at Massachusetts General Hospital in Boston and a member of the US National Academies Institute of Medicine. Dr. Weissleder’s research interests include the development of novel molecular imaging techniques, tools for early disease detection, and nanomaterials for systems analysis.
Author: Ralph Weissleder, Massachusetts General Hospital, Boston (USA), https://csb.mgh.harvard.edu/investigator/ralph_weissleder
Title: BODIPY—Tetrazine Derivatives as Superbright Bioorthogonal Turn-on Probes
Angewandte Chemie International Edition, Permalink to the article: http://dx.doi.org/10.1002/anie.201301100

Ralph Weissleder | Angewandte Chemie
Further information:
http://pressroom.angewandte.org

More articles from Life Sciences:

nachricht Unravelling the genetics of fungal fratricide
16.10.2018 | Uppsala University

nachricht Fungal weapon turns against the maker
16.10.2018 | Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie - Hans-Knöll-Institut (HKI)

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: Storage & Transport of highly volatile Gases made safer & cheaper by the use of “Kinetic Trapping"

Augsburg chemists present a new technology for compressing, storing and transporting highly volatile gases in porous frameworks/New prospects for gas-powered vehicles

Storage of highly volatile gases has always been a major technological challenge, not least for use in the automotive sector, for, for example, methane or...

Im Focus: Disrupting crystalline order to restore superfluidity

When we put water in a freezer, water molecules crystallize and form ice. This change from one phase of matter to another is called a phase transition. While this transition, and countless others that occur in nature, typically takes place at the same fixed conditions, such as the freezing point, one can ask how it can be influenced in a controlled way.

We are all familiar with such control of the freezing transition, as it is an essential ingredient in the art of making a sorbet or a slushy. To make a cold...

Im Focus: Micro energy harvesters for the Internet of Things

Fraunhofer IWS Dresden scientists print electronic layers with polymer ink

Thin organic layers provide machines and equipment with new functions. They enable, for example, tiny energy recuperators. In future, these will be installed...

Im Focus: Dynamik einzelner Proteine

Neue Messmethode erlaubt es Forschenden, die Bewegung von Molekülen lange und genau zu verfolgen

Das Zusammenspiel aus Struktur und Dynamik bestimmt die Funktion von Proteinen, den molekularen Werkzeugen der Zelle. Durch Fortschritte in der...

Im Focus: Dynamics of individual proteins

New measurement method allows researchers to precisely follow the movement of individual molecules over long periods of time

The function of proteins – the molecular tools of the cell – is governed by the interplay of their structure and dynamics. Advances in electron microscopy have...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

VideoLinks
Industry & Economy
Event News

Berlin5GWeek: Private industrial networks and temporary 5G connectivity islands

16.10.2018 | Event News

5th International Conference on Cellular Materials (CellMAT), Scientific Programme online

02.10.2018 | Event News

Major Project: The New Silk Road

01.10.2018 | Event News

 
Latest News

Unravelling the genetics of fungal fratricide

16.10.2018 | Life Sciences

Blue phosphorus -- mapped and measured for the first time

16.10.2018 | Physics and Astronomy

Berlin5GWeek: Private industrial networks and temporary 5G connectivity islands

16.10.2018 | Event News

VideoLinks
Science & Research
Overview of more VideoLinks >>>