Bacteriophage-derived nanopore sensors

Patent metadata

Field	Value
Publication number	US-11980849-B2
Application number	US-201916967560-A
Country	US
Kind code	B2
Filing date	Feb 8, 2019
Priority date	Feb 9, 2018
Publication date	May 14, 2024
Grant date	May 14, 2024

Abstract

Disclosed herein are compositions and methods that involve inserting connector protein channels of bacteriophage DNA packaging motors into copolymeric membranes via liposome-polymer fusion, which can be used as nanopore sensors for biomedical applications such as high throughput protein sequencing or cancer diagnosis. For example, disclosed are compositions comprising a copolymeric membrane into which a connector protein channel of a bacteriophage packaging motor has been inserted.

First claim

What is claimed is:

1. A copolymeric membrane comprising a nanopore inserted therein, wherein the nanopore is derived from a connector protein of a bacteriophage DNA packaging motor, wherein the nanopore comprises the aperture forming region of a connector protein of a bacteriophage DNA packaging motor, and wherein the aperture forming region is modified to alter one or more property of the channel of the nanopore.

2. The copolymeric membrane of claim 1 , wherein the connector protein of a bacteriophage DNA packaging motor is modified compared to the wild-type connector protein.

3. The copolymeric membrane of claim 1 , wherein the nanopore comprises a full length connector protein of a bacteriophage DNA packaging motor.

4. The copolymeric membrane of claim 1 , wherein the nanopore comprises a truncated connector protein of a bacteriophage DNA packaging motor.

5. The copolymeric membrane of claim 1 , wherein the nanopore is a multimeric protein formed of six or more subunits.

6. The copolymeric membrane of claim 5 , wherein the nanopore is a dodecameric protein.

7. The copolymeric membrane of claim 5 , wherein one or more of the subunits are modified at the C-terminus and/or N-terminus.

8. The copolymeric membrane of claim 7 , wherein one or more of the subunits are modified at the C-terminus and/or N-terminus to increase the hydrophilicity at one or both ends of the nanopore.

9. The copolymeric membrane of claim 7 , wherein one or more of the subunits are modified by the addition of a flexible linker and a peptide tag at the C-terminus and/or N-terminus.

10. The copolymeric membrane of claim 5 , wherein the subunits are identical.

11. The copolymeric membrane of claim 1 , wherein the bacteriophage DNA packaging motor is selected from the group consisting of phi29, T3, T4, T5, T7, SPP1, HK97, Lamda, G20c, P2, P3 and P22.

12. The copolymeric membrane of claim 1 , wherein the copolymeric membrane is a triblock or diblock copolymeric membrane.

13. An array of copolymeric membranes according to claim 1 .

14. The array of claim 13 , which is adapted for insertion into a device suitable for detecting the translocation of analytes through the nanopores in the array.

15. A device comprising an array of copolymeric membranes according to claim 1 .

16. A method of characterizing a target analyte, the method comprising applying a voltage potential across the copolymeric membrane of claim 1 , contacting the copolymeric membrane with the target analyte such that the target analyte moves with respect to the nanopore, and taking one or more measurements as the target analyte moves with respect to the pore, thereby determining the presence, absence or one or more characteristics of the analyte.

Assignees

• Ohio State Innovation Foundation
• Oxford Nanopore Tech Ltd

Inventors

• Guo Peixuan
• Ji Zhouxiang
• Jayasinghe Lakmal
• Jordan Michael

Classifications

• B01D69/144 Primary

  containing embedded or bound biomolecules · CPC title

• B01D67/0081

  After-treatment of organic or inorganic membranes · CPC title

• B01D71/74

  Natural macromolecular material or derivatives thereof (B01D71/08, B01D71/24 take precedence) · CPC title

• B01D71/80

  Block polymers · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title