Protein reporters for ultrasensitive detection methods

Patent metadata

Field	Value
Publication number	US-11300640-B2
Application number	US-201815919566-A
Country	US
Kind code	B2
Filing date	Mar 13, 2018
Priority date	Mar 13, 2017
Publication date	Apr 12, 2022
Grant date	Apr 12, 2022

Abstract

The present invention relates to compositions and methods using protein reporters as imaging agents in 129Xe NMR and MRI applications. It is described that bla and MBP are genetically-encoded proteins that induce a detectable chemical shift during 129Xe NMR, allowing for use as protein reporters in research and clinical applications.

First claim

What is claimed is:

1. An 129Xe NMR imaging agent comprising one or more protein reporters selected from the group consisting of TEM-1 β-lactamase (bla), and a mutant of bla, wherein the mutant of bla comprises a point mutation located at a residue selected from the group consisting of 1263, 1279, 1282, M182 and a combination thereof.

2. The imaging agent of claim 1 , wherein the one or more protein reporters comprise an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6.

3. The imaging agent of claim 1 , wherein the one or more protein reporters comprise a targeting domain that binds to a biomolecule of interest.

4. A method of 129Xe NMR imaging comprising the steps of:
a. administering to a cell or subject a 129Xe NMR imaging agent comprising one or more protein reporters selected from the group consisting of bla and a mutant of bla, wherein the mutant of bla comprises a point mutation located at a residue selected from the group consisting of I263, I279, I282, M182 and a combination thereof;
b. administering hyperpolarized xenon to the cell or subject;
c. obtaining magnetic resonance data of a target site of the cell or subject; and
d. analyzing the data to produce a magnetic resonance image of the target site.

5. The method of claim 4 , wherein the method further comprises administering a saturating radio frequency pulse to the target site.

6. The method of claim 4 , wherein the data comprises a chemical shift of about 60 ppm indicating the presence and location of said bla or said mutant of bla at the target site.

7. The method of claim 4 , wherein the protein reporter is administered at a concentration in the range of about 0.001 μM 100 μM.

8. The method of claim 4 , wherein the cell is in an in vitro or ex vivo environment.

9. The method of claim 4 , wherein the subject is a mammal.

10. The method of claim 4 , wherein the protein reporter further comprises a targeting domain that binds a biomolecule of interest.

11. The method of claim 4 , wherein the method is used to detect the presence of a tumor in the subject.

Assignees

• Univ Pennsylvania

Inventors

• Dmochowski Ivan J
• Wang Yanfei
• Roose Benjamin W
• Zemerov Serge
• Zhao Zhuangyu

Classifications

• C01B23/0005

  Compounds of noble gases · CPC title

• G01R33/282 Primary

  Means specially adapted for hyperpolarisation or for hyperpolarised contrast agents, e.g. for the generation of hyperpolarised gases using optical pumping cells, for storing hyperpolarised contrast agents or for the determination of the polarisation of a hyperpolarised contrast agent · CPC title

• G01R33/62

  using double resonance (G01R33/24 takes precedence) · CPC title

• A61K49/14

  Peptides, e.g. proteins · CPC title

• G16H30/40

  for processing medical images, e.g. editing · CPC title