HPLC free purification of peptides by the use of new capping and capture reagents

Patent metadata

Field	Value
Publication number	US-12415831-B2
Application number	US-202017419073-A
Country	US
Kind code	B2
Filing date	Jan 3, 2020
Priority date	Jan 7, 2019
Publication date	Sep 16, 2025
Grant date	Sep 16, 2025

Abstract

The present disclosure relates to the use of a capping and capture reagent in solid phase peptide synthesis. The present disclosure further relates to a method of solid phase peptide synthesis, wherein a capping and capture reagent according to the present disclosure is used. The present disclosure further relates to a method for purification of a (full-length) synthetic peptide via use of a capping and capture reagent according to the present disclosure. The present disclosure also relates to a kit comprising a capping and capture reagent according to the present disclosure and an amino oxy resin or a hydrazine resin and the use of the kit.

First claim

The invention claimed is:

1. A method for purification of a synthetic peptide comprising the steps of:
performing a solid phase peptide synthesis (SPPS), wherein individual amino acids are coupled to each other via Fmoc-chemistry, wherein after at least one coupling step a capping and capture reagent is used for capping a failure peptide sequence, said capping and capture (cap-capt) reagent comprising a capping moiety and a capture moiety, and wherein said capping moiety is a carboxylic acid or an activated carboxylic acid and wherein said capture moiety is a protected carbonyl group, the capping and capture reagent being selected from

 and

cleaving the compounds thus synthesized from the solid phase by adding trifluoroacetic acid (TFA) or TFA combined with one or more scavengers, wherein said one or more scavengers are selected from water, a silane, a thiol, a sulfide, and/or an ether,
deprotecting the capture moiety using trifluoroacetic acid (TFA) or TFA combined with one or more scavengers, wherein said one or more scavengers are selected from water, a silane, a thiol, a sulfide, and/or an ether, and
binding the deprotected capture moiety to a solid resin support thereby binding the at least one failure peptide sequence to the solid resin support and
separating the synthetic peptide from the solid resin support comprising the failure peptide.

2. Method according to claim 1 , wherein the solid resin support is an amino oxy resin, or hydrazine resin.

3. Method according to claim 1 , wherein the separating of the peptide from the solid resin support comprising the failure peptide is carried out by centrifugation or by filtration.

4. The method according to claim 1 , wherein the one or more scavengers is selected from triethylsilane, triisopropylsilane, mercaptoethanol, dithioerythritol, ethanedithiol, dioxa-1,8-octane-dithiol, thioanisole, ethylmethylsulfide, and/or anisole.

Assignees

• Roche Diagnostics Operations Inc

Inventors

• Bergmann Frank
• Pomplun Sebastian Johannes

Classifications

• C07K1/04

  on carriers {(C07K1/003, C07K1/006 take precedence)} · CPC title

• C07K1/20 Primary

  Partition-, reverse-phase or hydrophobic interaction chromatography · CPC title

• C07K1/063 Primary

  for alpha-amino functions · CPC title