Affinity tags and processes for purifying and immobilizing proteins using same

Patent metadata

Field	Value
Publication number	US-2016159856-A1
Application number	US-201414904029-A
Country	US
Kind code	A1
Filing date	Sep 19, 2014
Priority date	Sep 19, 2013
Publication date	Jun 9, 2016
Grant date	—

Abstract

The present disclosure provides affinity tags, fusion proteins comprising one or more affinity tags, compositions comprising a fusion protein, methods of purifying a protein using an affinity tag, and devices for purifying a protein using an affinity tag.

First claim

1 . A protein comprising SEQ ID NO.: 1.

2 . A protein comprising SEQ ID NO.: 2

3 . A protein comprising SEQ ID NO.: 3

4 . A protein comprising SEQ ID NO.: 4

5 . A protein comprising an amino acid sequence having a homology of at least 70%, at least 80%, at least 90%, or at least 95% with the amino acid sequence of any one of SEQ ID NOs.: 1 to 4.

6 . The protein of claim 5 , wherein the protein is a fusion protein.

7 . The protein of claim 6 , wherein the fusion protein comprises a polyclonal antibody, a monoclonal antibody, a complement determining region-grafted antibody preparation, a hybrid antibody, an altered antibody, a F(ab)′ 2 fragment, a Fab molecule, a Fv fragment, a single domain antibody, a chimeric antibody, or a fragment thereof.

8 . A substrate comprising a bound protein of claim 1 .

9 . The substrate of claim 8 , wherein the substrate includes a surface comprising a silicon oxide.

10 . The substrate of claim 8 , wherein the substrate comprises silica.

11 . A method of isolating a protein, the method comprising:
contacting a silicon oxide substrate with a fluid comprising the protein for a time sufficient to allow the protein to bind to the silicon oxide substrate; contacting the silicon oxide substrate with a washing fluid; and contacting the silicon oxide substrate with a releasing agent to release the bound protein.

12 . The method of claim 11 , wherein the protein is a protein of any one of claims 1 to 7 .

13 . The method of claim 11 , wherein the releasing agent comprises lysine and/or arginine.

14 . The method of claim 11 , wherein the releasing agent comprises a protease.

15 . The method of claim 11 , wherein the silicon oxide substrate comprises silica, borosilicate, controlled pore glass, quartz, and/or oxidized silicon.

16 . The method of claim 12 , further comprising contacting the released protein with a protease to cleave the SEQ ID NOs.: 1 to 4, or homologous portion thereof, from the protein.

17 . A kit for purifying the protein of claim 1 , the kit comprising;
a silicon oxide substrate; a vessel for retainably holding the silicon oxide substrate; a receptacle for receiving eluent passing through the vessel; and a releasing agent.

18 . The kit of claim 17 further comprising instructions for using the kit to purify the protein.

19 . The kit of claim 17 , wherein the releasing agent comprises arginine and/or lysine.

20 . The kit of claim 17 , wherein the releasing agent comprises a protease.

21 - 27 . (canceled)

Assignees

• Univ Washington Ct Commerciali

Inventors

• Baaneyx Francois C
• Coyle Brandon L

Classifications

• C07K1/20

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

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• C07K7/08

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title

• C07K2319/20

  containing a tag with affinity for a non-protein ligand · CPC title

• C07K1/22 Primary

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title