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 Desirable defects
30.04.2015 | International School of Advanced Studies (SISSA)

nachricht Rare Dune Plants Thrive on Disturbance
30.04.2015 | Washington University in St. Louis

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Erosion, landslides and monsoon across the Himalaya

Scientists from Nepal, Switzerland and Germany was now able to show how erosion processes caused by the monsoon are mirrored in the sediment load of a river crossing the Himalaya.

In these days, it was again tragically demonstrated that the Himalayas are one of the most active geodynamic regions of the world. Landslides belong to the...

Im Focus: Through the galaxy by taxi - The Dream Chaser Space Utility Vehicle

A world-class prime systems integrator and electronic systems provider known for its rapid, innovative, and agile technology solutions, Sierra Nevada Corporation (SNC) is currently developing a new space transportation system called the Dream Chaser.

The ultimate aim is to construct a multi-mission-capable space utility vehicle, while accelerating the overall development process for this critical capability...

Im Focus: High-tech textiles – more than just clothes

Today, textiles are used for more than just clothes or bags – they are high tech materials for high-tech applications. High-tech textiles must fulfill a number of functions and meet many requirements. That is why the Fraunhofer Institute for Silicate Research ISC dedicated some major developing work to this most intriguing research area. The result can now be seen at Techtextil trade show in Frankfurt from 4 to 7 May. On display will be novel textile-integrated sensors, a unique multifunctional coating system for textiles and fibers, and textile processing of glass, carbon, and ceramics fibers to fiber preforms.

Thin materials and new kinds of sensors now make it possible to integrate silicone elastomer sensors in textiles. They are suitable for applications in medical...

Im Focus: Fast and Accurate 3-D Imaging Technique to Track Optically-Trapped Particles

KAIST researchers published an article on the development of a novel technique to precisely track the 3-D positions of optically-trapped particles having complicated geometry in high speed in the April 2015 issue of Optica.

Daejeon, Republic of Korea, April 23, 2015--Optical tweezers have been used as an invaluable tool for exerting micro-scale force on microscopic particles and...

Im Focus: NOAA, Tulane identify second possible specimen of 'pocket shark' ever found

Pocket sharks are among the world's rarest finds

A very small and rare species of shark is swimming its way through scientific literature. But don't worry, the chances of this inches-long vertebrate biting...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

HHL Energy Conference on May 11/12, 2015: Students Discuss about Decentralized Energy

23.04.2015 | Event News

“Developing our cities, preserving our planet”: Nobel Laureates gather for the first time in Asia

23.04.2015 | Event News

HHL's Entrepreneurship Conference on FinTech

13.04.2015 | Event News

 
Latest News

Dust from the Sahara Desert cools the Iberian Peninsula

30.04.2015 | Earth Sciences

Desirable defects

30.04.2015 | Life Sciences

Germany's DanTysk Offshore Wind Power Plant Inaugurated

30.04.2015 | Press release

VideoLinks
B2B-VideoLinks
More VideoLinks >>>