Fluorous Peptide Synthesis

Benefits of fluorous techniques in peptide synthesis

  • Streamline your peptide synthesis workflow
  • Minimize purification method development by using a single general purification technique
  • Increase final HPLC purification efficiency through fluorous pre-purification
  • Easily remove deletion sequences in solid phase peptide synthesis

Fluorous techniques can simplify and streamline your peptide synthesis and purification whether you are making peptide libraries in small quantities or single therapeutic peptides in large quantity. The key to the purification is fluorous solid phase extraction (FSPE), a single general purification technique that is applicable over a wide range of peptides. Since FSPE only differentiates between fluorous tagged and non-tagged entities, an entire library of peptides can be purified using a single method. If you are making a single peptide the same method can be used to separate closely related peptides, such as deletion sequences, to provide an efficient and simple method by which to pre-purify a sample prior to final reverse phase HPLC resulting in greater recovery and efficiency.

Fluorous Techniques in Solid Phase Peptide Synthesis

Fluorous Technologies Inc. has developed a superior fluorous Fmoc reagent for N-terminal tagging that offers clear advantages over other fluorous tags. The basic methodology uses Fmoc-based solid-phase chemistry to build the amino acid chain. After each coupling step, any free amines from deletions are capped with an acetate group. After the desired number of iterations and thefinal Fmoc deprotection, the fluorous Fmoc is added to fluorous tag the desired full sequence. The peptides are easily cleaved from the resin to provide a mixture of the desired fluorous-tagged peptide and undesired truncated sequences, which do not contain a fluorous tag.

The fluorous-tagged material is then easily separated from the non-tagged material by fluorous solid phase extraction (FSPE) or fluorous HPLC. Conversely, a fluorous capping strategy can also be employed where all the truncated sequences are fluorous and the desired full sequence is non-fluorous [2].

Fluorous Techniques in Solution Phase Synthesis

In solution phase syntheses, two approaches can be used: fluorous supported peptide synthesis[3] and fluorous supported amide coupling reagents[4]. In both instances all reaction are homogeneous resulting in less equivalents of reagents for each iteration, favorable reaction kinetics, and full reaction monitoring using standard analytical techniques. The fluorous tagged components can be separated from the non-tagged components using either FSPE or fluorous liquid-liquid extraction (FLLE).

Peptide Synthesis

"Immobilization-Ready" when used with fluorous-modified glass slides

Immobilize your newly synthesized peptides without changing the tag. The unique properties of fluorous compounds allow them to be immobilized onto fluorous-modified surfaces in the formation of microarrays. To date, small molecules, peptides, and carbohydrate arrays have all been formed using fluorous immobilization. Read more about fluorous peptide microarrays here.

Fluorous Technologies Inc. offers a complete line of fluorous tags, reagents, and separation media for peptide synthesis and purification. Fluorous techniques can be used in conjunction with solid-phase peptide synthesis (SPPS) or in solution phase synthesis.

Selected References

  • a) Filippov, D. V.; van Zoelen, D. J.; Oldfield, S. P.; van der Marel, G. A.; Overkleeft, H. S.; Drijfhout, J. W.; van Boom, J. H., Tetrahedron Letters 2002, 43, (43), 7809-7812.
    b) de Visser, P. C.; van Helden, M.; Filippov, D. V.; van der Marel, G. A.; Drijfhout, J. W.; van Boom, J. H.; Noort, D.; Overkleeft, H. S., Tetrahedron Letters 2003, 44, (50), 9013-9016.
  • Montanari, V.; Kumar, K., Just Add Water: Journal of the American Chemical Society 2004, 126, (31), 9528-9529.
  • Goto, K.; Miura, T.; Mizuno, M., Tetrahedron Letters 2005, 46, (48), 8293-8297.
  • Matsugi, M.; Hasegawa, M.; Sadachika, D.; Okamoto, S.; Tomioka, M.; Ikeya, Y.; Masuyama, A.; Mori, Y. Tetrahedron Lett. 2007, 48, 4147-4150.