Surface plasmon resonance approach to monitor protein-ligand interactions

Patent metadata

Field	Value
Publication number	US-10429300-B2
Application number	US-201615545980-A
Country	US
Kind code	B2
Filing date	Jan 26, 2016
Priority date	Jan 26, 2015
Publication date	Oct 1, 2019
Grant date	Oct 1, 2019

Abstract

The present invention provides assays utilizing SPR to detect protein-ligand interactions as well as compositions utilized is such assays.

First claim

What is claimed is:

1. A method of identifying a ligand that binds to a paddle motif peptide, the method comprising:
a) contacting a paddle motif peptide immobilized on a sensor chip with a test molecule, wherein the paddle motif peptide comprises SEQ ID NO: 2 or SEQ ID NO: 4;
b) detecting binding of the ligand and the paddle motif peptide using surface plasmon resonance (SPR), thereby identifying the test molecule as a ligand that binds the paddle motif peptide.

2. The method of claim 1 , wherein the sensor chip comprises multiple paddle motif peptides, each comprising a voltage-sensing domain (VSD) of a voltage-gated channel protein.

3. The method of claim 2 , wherein the voltage-gated channel protein is selected from the group consisting of a sodium (Nav) channel protein, a potassium (Kv) channel protein, and a calcium (Cav) channel protein.

4. The method of claim 1 , wherein the test molecule is a peptide.

5. The method of claim 4 , wherein the peptide is an antibody, or fragment thereof.

6. The method of claim 1 , wherein the method is a label free optical assay.

7. The method of claim 1 , further comprising generating a sensorgram.

8. The method of claim 1 , further comprising calculating a kinetic rate constant.

9. A sensor chip for use in a surface plasmon resonance (SPR) assay, the chip comprising a paddle motif peptide immobilized thereon, wherein the paddle motif peptide comprises SEQ ID NO: 2 or SEQ ID NO: 4.

10. The chip of claim 9 , wherein the sensor chip comprises multiple paddle motif peptides, each comprising a voltage-sensing domain (VSD) of a voltage-gated channel protein.

11. The chip of claim 10 , wherein the voltage-gated channel protein is selected from the group consisting of a sodium (Nav) channel protein, a potassium (Kv) channel protein, and a calcium (Cav) channel protein.

12. A paddle motif peptide comprising SEQ ID NO: 2.

13. The peptide of claim 12 , wherein the peptide consists of SEQ ID NO: 2.

14. The peptide of claim 12 , wherein the peptide is biotinylated.

15. A paddle motif peptide comprising SEQ ID NO: 4.

16. The peptide of claim 15 , wherein the peptide consists of SEQ ID NO: 2.

17. The peptide of claim 15 , wherein the peptide is biotinylated.

18. A kit comprising the sensor chip of claim 9 , reagents, and instructions for conducting a surface plasmon resonance (SPR) assay with the sensor chip.

Assignees

• Univ Johns Hopkins
• Centre Nat Rech Scient
• Aix Marseilles Univ
• Centre Nat Rech Scient

Inventors

• Bosmans Frank
• Bougis Pierre E
• Eauclaire Marie-France

Classifications

• C12N9/1205

  Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases · CPC title

• G01N21/553 Primary

  and using surface plasmons (fluorescence excitation G01N21/648; enhanced Raman G01N21/658) · CPC title

• G01N33/54373

  involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings · CPC title

• C07K14/58

  Atrial natriuretic factor complex; Atriopeptin; Atrial natriuretic peptide [ANP]; Cardionatrin; Cardiodilatin · CPC title

• G01N33/6872

  Intracellular protein regulatory factors and their receptors, e.g. including ion channels · CPC title