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CEOverture...
Welcome to the May edition of our Newsletter. In this edition we highlight several new products for use in making, purifying and immobilizing molecules. If you haven’t checked in on fluorous chemistry in awhile, now is a great time to get back up to speed, as the technology continues to evolve at a rapid pace. Continuous innovation is our goal, and hopefully you will find the new products useful in your daily work. Help us bring to market the most useful products by sending suggestions to contact-fti@fluorous.com. We always enjoy hearing from you.
Cheers,
Phil
Philip E. Yeske
President & CEO
- FluoroFlash® 5 μm silica formatted in pipette Tips and LC trap cartridges for proteomics applications
- Fluorous Modified Glass Slides for Microarray formation
- F-DCT and a new Fluorous Mukaiyama Reagent for amide couplings
- FluoroFlash® silica gel in 96- Well Plates for parallel synthesis
Beta Products Now Available for Fluorous Proteomics & Microarrays
The application of fluorous tags for the enrichment of specific peptide subsets from a biological sample has been reported by Brittain et al (1). Since that report, FTI has been offering to proteomics researchers fluorous tagging reagents with various reactive functionalities, such as a NHS ester for amine tagging and a maleimide for Cys tagging. After reaction with the appropriate reagent, the tagged peptides are separated from non-tagged species using FTI’s 5 μm FluoroFlash® silica gel. FTI is happy to announce that we are now providing the 5 μm FluoroFlash® silica gel formatted either in pipette tips for off-line enrichment or LC trap cartridges for on-line purification as beta products. The pipette tips are available in two sizes, 1-10μL and 10-200μL (catalog 801-5040, 801-5050) as are the LC trap cartridges, 40 μL and 100 μL bed volume sizes. Each can be used for the selective enrichment of fluorous species from non-fluorous species.
Beyond synthesis and separation, fluorous tagged molecules have also been effectively immobilized on a fluorous modified glass slide in the formation of microarrays (2). Fluorous microarrays are marked by the following features:
- High sensitivity
- High signal-to-noise
- Low background fluorescence
- Low non-specific binding allowing incubation with cell lysates
- Simplified workflows, due to the elimination of blocking and washing steps
While exact protocols are still under development for various molecular classes, initial beta-testers have provided overwhelmingly positive feedback regarding the performance of the slides. Fluorous Technologies can now provide the fluorous modified glass slides (catalog 850-9100) to researchers looking for new surface chemistries that provide superior performance over existing microarray technologies.
If you would like to be among the first to try any of these new products, please contact us and we will be happy to provide you with all of the details.
References:
1. Brittain, S. M.; Ficarro, S. B.; Brock, A.; Peters, E. C. Nature Biotechnology 2005, 23, 463-468.
2. Ko, K.-S.; Jaipuri, F. A.; Pohl, N. L. Journal of the American Chemical Society 2005, 127, 13162-13163.
On-Target Fluorous Enrichment & Direct MS Analysis
Prof. Gary Siuzdak of The Scripps Research Institute and Eric Peters of the Genomics Institute of the Novartis Research Foundation recently disclosed their results in on-target enrichment studies of fluorous modified molecules (1). The researchers described the fluorous tagging of specific functional groups on amino acids, carbohydrates, and peptides in complex mixtures. These mixtures are deposited onto a fluorous modified DIOS target which is then washed to remove non-fluorous species and enrich the target in the fluorous tagged species. Analysis of the deposited samples were then conducted through desorption by irradiation of the target and direct MS analysis. Importantly, the analysis could occur both before and after washing, thanks to the selectivity of fluorous solid phase extraction. After performing the on-target enrichment and analysis on various simple mixtures, the researchers also demonstrated the fluorous tagging at the protein level followed by all the standard gel-based manipulations (SDS-PAGE, in-gel digestion, etc.). The extracted peptides were then deposited on a fluorous DIOS target and analyzed. Finally, the researchers analyzed amino acids found in human serum and found that the fluorous esterification of carboxylic acids using 3-perfluorobutyl-propanol (catalog F009029) and the on-target enrichment approach resulted in greater sensitivity than found using a more classic non-fluorous esterification technique. The work described provides another example of the power of fluorous based separations in analytical applications.
References:
1. Go, E.P.; Uritboonthai, W.; Apon, J.V.; Trauger, S.A.; Nordstrom, A.; O’Maille, G.; Brittain, S.M.; Peters, E.C.; Siuzdak, G., Journal of Proteome Research, 2007, 6, 1492.
Fluorous Dichlorotriazine (F-DCT) for Amide Coupling & Nucleophile Scavenging
As an analog of 2-chlroro-4,6-dimethoxytriazine (CDMT), 2,4-dichloro-6-(perfluorooctyl)-propyloxy-1,3,5-triazine (F-DCT) has been developed as a coupling reagent for amidation reactions and peptide synthesis.(1) In the presence of N-methylmorpholine (NMM), the carboxylic acid activated by F-DCT is coupled with an amine to from the C-N bond. Since F-DCT has two active chlorines, it leads to formation of two equivalents of product. This reagent has also be used as a scavenger to remove excess nucleophiles such as amines and thiols from the reaction mixture (2). Just like in other fluorous reactions, F-DCT reactions have distinct advantages, including solution-phase kinetics, the ability to monitor reactions using standard techniques, and the facile removal of fluorous by-products using fluorous solid phase extraction (F-SPE). Please contact FTI for pricing and availability of this new reagent.
References:
1. Zhang, W.; Lu, Y. QSAR Comb. Sci. 2006, 25, 724-727
2. Lu, Y.; Zhang, W. QSAR Comb. Sci. 2006, 25, 728-731.
Light Fluorous Mukaiyama Reagent
Mukaiyama’s condensation reagent 1 has been widely used in the formation of esters and amides. Recently, a new fluorous Mukaiyama reagent 2 has been reported by Prof. Matsugi’s group in Japan (1). The researchers have shown that 2 has comparable reactivity to 1, but with the advantage that the pyridone by-product can be efficiently removed by F-SPE using 80:20 MeOH-H2O as the organic wash. As shown below, the reaction between 4-methoxybenzoic acid and N-methylaniline gave 77% yield of the amide product in excellent purity (98.9% by HPLC) after aqueous workup and F-SPE. Matsugi and co-workers reported a variety of esters and amides which were formed using the fluorous Mukaiyama reagent. Compared to earlier fluorous versions of 1, compound 2 is more reactive, does not require HOBT as a co-reagent, and is more cost effective. All of these features should make this new fluorous Mukaiyama reagent an excellent choice for the production of amide and ester libraries. Please contact FTI for pricing and availability of this new reagent.
References:
1. Matsugi, M., Hasegawa, M., Sadachika, D., Okamoto, S., Tomioka, M., Ikeya, Y.,
Masuyama, A., Mori, Y., Tetrahedron Letters 2007, DOI: 10.1016/j.tetlet.2007.03.173
96-Well Gravity F-SPE Plates for High-Throughput Purification
In our continuing effort to increase the capabilities and value of fluorous techniques, FTI is introducing a gravity-driven 96-well plate-to-plate F-SPE as a follow-up to 24-well and automated F-SPE (1) processes commercialized last year. 
By using a 96-well plate (catalog 807-0096), up to 50 mg of reaction mixture can be loaded into each well of the plate, which is then washed with a fluorophobic solvent such as DMSO, DMF or 85:15 DMF-H2O, followed by a fluorophilic wash using MeOH or THF. The desired products are collected in the receiving plates and directly concentrated in a vacuum centrifuge. This simple and highly efficient parallel separation technique has been demonstrated in the purification of four 96-compound libraries produced by scavenging reactions with 1-(perfluoroctyl)propyl isatoic anhydride (catalog F017028), amide coupling reactions with 2-chloro-4,6-bis[(perfluorooctyl)propyloxy]-1,3,5-triazine (F-CDMT, catalog F026171) or 2,4-dichloro-6-(perfluorooctyl)propyloxy-1,3,5-triazine (F-DCT, catalog XP017178), and Mitsunobu reactions with fluorous diethyl azodicarboxylate (F-DIAD, catalog F026100) and triphenylphosphine (F-TPP, catalog F017039). Approximately 80% of the products in each library have greater than 85% purity after F-SPE without the need to conduct additional chromatography. A manuscript describing details of the actual libraries made has been submitted for publication, but in the meantime you can find the protocol used in the Application Note. The extension of fluorous techniques into 96 well plate format increases overall throughput allowing you to be even more efficient in your work.
References:
1. Zhang, W.; Lu, Y. J. Comb. Chem. 2006, 8, 890-896.
Fluorous Technologies Event Page
We have recently updated our webpage for upcoming events where you can find out where scientists utilizing fluorous techniques will be presenting oral presentations or posters related to recent developments in fluorous chemistry. If you are planning on attending one of these events then be sure to contact us to set up a time to get together to learn more about how fluorous techniques can help you succeed.
Are you a fluorous researcher planning on giving an oral presentation or poster at an upcoming meeting? We would gladly include you on our events page. Just send us the name and location of the event, the title and presenter of the talk or poster to add to our rapidly growing list of leaders in fluorous research.
ISoFT07 - See you in Japan!
FTI is a proud sponsor of the 2nd International Symposium on Fluorous Technologies (ISoFT07), to be held in Yokohama, Japan between July 29th and August 1st, 2007. Following up on the success of ISoFT05 (here is the link http://isoft05.free.fr/) in Bordeaux, France, this conference is sure to be a can’t miss event for anyone active in this rapidly expanding field. Hope to see you there!
Tides 2007, May 20-23, Las Vegas
FTI partner - Berry & Associates - will be present at the TIDES conference later this month. In addition to their Booth #202 in the Exhibit Hall, they will also present a poster entitled "Fluorous Purification of Oligonucleotides." Be sure to stop by and find out more about the value of fluorous technology in this important field.