Solid phase peptide synthesis processes and associated systems

Patent metadata

Field	Value
Publication number	US-9695214-B2
Application number	US-201414776870-A
Country	US
Kind code	B2
Filing date	Feb 24, 2014
Priority date	Mar 15, 2013
Publication date	Jul 4, 2017
Grant date	Jul 4, 2017

Abstract

Systems and processes for performing solid phase peptide synthesis are generally described. Solid phase peptide synthesis is a known process in which amino acid residues are added to peptides that have been immobilized on a solid support. In certain embodiments, the inventive systems and methods can be used to perform solid phase peptide synthesis quickly while maintaining high yields. Certain embodiments relate to processes and systems that may be used to heat, transport, and/or mix reagents in ways that reduce the amount of time required to perform solid phase peptide synthesis.

First claim

What is claimed is:

1. A process for adding amino acid residues to peptides, comprising:
providing a plurality of peptides comprising protection groups, each peptide immobilized on a solid support;
performing a first amino acid addition cycle comprising exposing the immobilized peptides to a first heated fluid stream comprising first activated amino acids such that a first amino acid residue is added to at least about 99% of the immobilized peptides, wherein the first activated amino acids are activated prior to and within 30 seconds of exposing the immobilized peptide to the first heated fluid stream and a pressure drop across the solid support does not exceed about 700 psi for more than about 5% of the time period during which the first amino acid addition cycle is performed; and
performing a second amino acid addition cycle comprising exposing the immobilized peptides to a second heated fluid stream comprising second activated amino acids such that a second amino acid residue is added to at least about 99% of the immobilized peptides, wherein the second activated amino acids are activated prior to and within 30 seconds of exposing the immobilized peptide to the second heated fluid stream and a pressure drop across the solid support does not exceed about 700 psi for more than about 5% of the time period during which the second amino acid addition cycle is performed;
wherein:
the total amount of time between the ends of the first and second amino acid addition cycles is about 10 minutes or less and the protection groups comprise fluorenylmethyloxycarbonyl protection groups and/or
the total amount of time between the ends of the first and second amino acid addition cycles is about 5 minutes or less.

2. A process for adding amino acid residues to peptides, comprising:
providing a plurality of peptides comprising protection groups, each peptide immobilized on a solid support;
exposing a deprotection reagent to the immobilized peptides to remove protection groups from at least a portion of the immobilized peptides;
removing at least a portion of the deprotection reagent;
exposing the immobilized peptides to a heated fluid stream comprising activated amino acids such that at least a portion of the activated amino acids are bonded to the immobilized peptides to form newly-bonded amino acid residues, wherein the activated amino acids are activated prior to and within 30 seconds of the exposing step, and wherein, during the exposing step, a pressure drop across the solid support does not exceed about 700 psi for more than about 5% of the time period during which the exposing step is performed; and
removing at least a portion of activated amino acids that do not bond to the immobilized peptides;
wherein an amino acid residue is added to at least about 99% of the immobilized peptides during the amino acids exposing step; and
wherein:
the total amount of time taken to perform the combination of all of the deprotection reagent exposing step, the deprotection reagent removal step, the activated amino acid exposing step, and the activated amino acid removal step is about 10 minutes or less and the protection groups comprise fluorenylmethyloxycarbonyl protection groups and/or
the total amount of time taken to perform the combination of all of the deprotection reagent exposing step, the deprotection reagent removal step, the activated amino acid exposing step, and the activated amino acid removal step is about 5 minutes or less.

3. A process for adding amino acid residues to peptides, comprising:
providing a plurality of peptides immobilized on a solid support; and
exposing the immobilized peptides to a heated fluid stream comprising activated amino acids such that at least a portion of the activated amino acids are bonded to the immobilized peptides to form newly-bonded amino acid residues,
wherein the activated amino acids are activated prior to and within 30 seconds of the exposing step,
wherein, during the exposing step, a pressure drop across the solid support does not exceed about 700 psi for more than about 5% of the time period during which the exposing step is performed; and
wherein an amino acid residue is added to at least about 99% of the immobilized peptides within about 1 minute or less.

4. The process of claim 3 , wherein an amino acid residue is added to at least about 99.9% of the immobilized peptides within about 1 minute or less.

5. The process of claim 3 , wherein an amino acid residue is added to at least about 99% of the immobilized peptides within about 30 seconds or less.

6. The process of claim 3 , wherein the solid support is contained within a packed column and/or a fluidized bed.

7. The process of claim 3 , wherein the solid support comprises polystyrene and/or polyethylene glycol.

8. The process of claim 3 , wherein the solid support comprises a resin.

9. The process of claim 8 , wherein the solid support comprises a microporous polystyrene resin.

10. The process of claim 3 , wherein multiple copies of the amino acid residue are bonded to fewer than about 1% of the immobilized peptides during the exposing step.

11. The process of claim 3 , wherein adding an amino acid residue to the immobilized peptides comprises adding a single amino acid residue to the immobilized peptides.

12. The process of claim 3 , wherein adding an amino acid residue to the immobilized peptides comprises adding a peptide comprising two or more amino acid residues to the immobilized peptides.

13. The process of claim 3 , wherein the heated fluid stream has a temperature that is at least about 10° C. greater than room temperature.

Assignees

• Massachusetts Inst Technology

Inventors

• Simon Mark David
• Pentelute Bradley L
• Adamo Andrea
• Heider Patrick Louis
• Jensen Klavs F

Classifications

• C07K1/045

  using devices to improve synthesis, e.g. reactors, special vessels · CPC title

• C07K1/061

  using protecting groups · CPC title

• C07K1/042 Primary

  characterised by the nature of the carrier · CPC title

• C07K1/08

  using activating agents {(C07K1/003, C07K1/006 take precedence)} · CPC title