IP Library Granted Patent US 10,150,984
Granted Patent B2
US 10,150,984 · App. 15/358,320 · Granted Dec 11, 2018

Tyrosine kinase biosensors and methods of use

Inventors: Laurie L Parker (Minneapolis, MN); Andrew Michael Lipchik (Palo Alto, CA); Scott Charles Bolton (West Lafayette, IN)
C12Q1/485
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Quick Facts
Patent No.
US 10,150,984
App. No.
15/358,320
Granted
Dec 11, 2018
Kind
B2
Abstract

Disclosed are compositions and methods for measuring tyrosine kinase activity.

Claims (9)

1. A biosensor comprising:

a peptide substrate that includes a sequence selected from the group consisting of ELDAYLENE (SEQ ID NO:14), ELAGYLENE (SEQ ID NO:15), ELDVYEEQL (SEQ ID NO:16), ELDVYVEQT (SEQ ID NO:17), DEDIYEELD (SEQ ID NO:18), EGDVYDFVE (SEQ ID NO:19), NNDVYEQPE (SEQ ID NO:20), EEDVYDMPD (SEQ ID NO:21), EADVYDMPD (SEQ ID NO:22), DLDIYEELD (SEQ ID NO:23), EAHVYDMMD (SEQ ID NO:24), DPDRYIRTE (SEQ ID NO:25), EGDRYLKLE (SEQ ID NO:26), EDGRYVQLD (SEQ ID NO:27), PKPRYVQLD (SEQ ID NO:28), DEDVYEELD (SEQ ID NO:38), DEDDYEDID (SEQ ID NO:42), DKDIYEELD (SEQ ID NO:43), DEDDYGDVD (SEQ ID NO:44), GGEEDEDIYEELDEPGGK biotin GG (SEQ ID NO:45), and GGDNEGDVYDFVEDGGK biotin GG (SEQ ID NO:46), the substrate specific for a tyrosine kinase selected from the group consisting of Btk, Src family kinases, and Jak2; and at least one of a cell penetrating peptide and an affinity tag, the cell penetrating peptide and/or tag linked directly or indirectly to the peptide substrate.

2. The biosensor of claim 1 , wherein the peptide substrate includes a sequence selected from the group consisting of ELDAYLENE (SEQ ID NO:14), ELAGYLENE (SEQ ID NO:15), ELDVYEEQL (SEQ ID NO:16), and ELDVYVEQT (SEQ ID NO:17).

3. The biosensor of claim 1 , wherein the peptide substrate includes a sequence selected from the group consisting of DEDIYEELD (SEQ ID NO:18), EGDVYDFVE (SEQ ID NO:19), NNDVYEQPE (SEQ ID NO:20), EEDVYDMPD (SEQ ID NO:21), EADVYDMPD (SEQ ID NO:22), DLDIYEELD (SEQ ID NO:23), and EAHVYDMMD (SEQ ID NO:24).

4. The biosensor of claim 1 , wherein the peptide substrate includes a sequence selected from the group consisting of DPDRYIRTE (SEQ ID NO:25), EGDRYLKLE (SEQ ID NO:26), EDGRYVQLD (SEQ ID NO:27), and PKPRYVQLD (SEQ ID NO:28).

5. The biosensor of claim 1 , wherein the biosensor comprises the sequence GGEEDEDIYEELDEPGGK biotin GG (SEQ ID NO:45).

6. The biosensor of claim 1 , wherein the biosensor comprises the sequence GGDNEGDVYDFVEDGGK biotin GG (SEQ ID NO:46).

7. The biosensor of claim 1 , wherein the cell penetrating peptide includes a sequence selected from the group consisting of GRKKRRQRRRPPQ (SEQ ID NO:47), RKKRRQRRR (SEQ ID NO:35), RQIKIWFQNRRMKWKK (SEQ ID NO:48), GWTLNSAGYLLGKINLKALAALAKKIL (SEQ ID NO:49), GALFLGFLGAAGSTMGAWSQPKKKRKV (SEQ ID NO:50), GALFLGFLGAAGSTMGA (SEQ ID NO:51), and EQNPIYWARYADWLFTTPLLLLDLALLVDADEGT (SEQ ID NO:52).

8. The biosensor of claim 1 , wherein the affinity tag includes a moiety selected from the group consisting of biotin and His6 chemical groups.

Assignments (1)
CONFIRMATORY LICENSE Recorded Sep 12, 2017
From: PURDUE UNIVERSITY
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 043820/0737 →
Continuity (8)
Division 13761968 · Feb 7, 2013
Provisional Application 61736312 · Dec 12, 2012
Provisional Application 61704298 · Sep 21, 2012
Provisional Application 61693002 · Aug 24, 2012
Provisional Application 61605591 · Mar 1, 2012
Provisional Application 61603752 · Feb 27, 2012
Provisional Application 61595959 · Feb 7, 2012
Related Publication 20170342461A1 · Nov 30, 2017