Granted Patent B1

US 12,421,306 · App. 17/953,930 · Granted Sep 23, 2025

Inventors: Brooke Nicole Harmon (Livermore, CA); Maxwell Stefan (Pleasanton, CA); Jennifer Schwedler (Livermore, CA); Yooli Kim Light (Pleasanton, CA); Catherine Margaret Mageeney (Livermore, CA)

Assignee: National Technology & Engineering Solutions of Sandia, LLC

C07K16/18A61P11/00A61K2039/505C07K2317/22C07K2317/569

Abstract

Claims (105)

1. A single-domain antibody construct, comprising a first single-domain antibody with a binding domain, wherein the binding domain comprises:

a first polypeptide comprising first, second, and third complementarity determining regions selected from the group consisting of the following combinations:

SEQ ID NOs: 1, 26, and 51, respectively;

SEQ ID NOs: 2, 27, and 52, respectively;

SEQ ID NOs: 3, 28, and 53, respectively;

SEQ ID NOs: 4, 29, and 54, respectively;

SEQ ID NOs: 5, 30, and 55, respectively;

SEQ ID NOs: 6, 31, and 56, respectively;

SEQ ID NOs: 7, 32, and 57, respectively;

SEQ ID NOs: 8, 33, and 58, respectively;

SEQ ID NOs: 9, 34, and 59, respectively;

SEQ ID NOs: 10, 35, and 60, respectively;

SEQ ID NOs: 11, 36, and 61, respectively;

SEQ ID NOs: 12, 37, and 62, respectively;

SEQ ID NOs: 13, 38, and 63, respectively;

SEQ ID NOs: 14, 39, and 64, respectively;

SEQ ID NOs: 15, 40, and 65, respectively;

SEQ ID NOs: 16, 41, and 66, respectively;

SEQ ID NOs: 17, 42, and 67, respectively;

SEQ ID NOs: 18, 43, and 68, respectively;

SEQ ID NOs: 19, 44, and 69, respectively;

SEQ ID NOs: 20, 45, and 70, respectively;

SEQ ID NOs: 21, 46, and 71, respectively;

SEQ ID NOs: 22, 47, and 72, respectively;

SEQ ID NOs: 23, 48, and 73, respectively;

SEQ ID NOs: 24, 49, and 74, respectively; and

SEQ ID NOs: 25, 50, and 75, respectively.

2. The single-domain antibody construct of claim 1 , further comprising one or more therapeutic or diagnostic agents.

3. The single-domain antibody construct of claim 1 , further comprising a therapeutic agent selected from the group consisting of: a therapeutic antibody, a small molecule drug, a chemotherapeutic agent, an antiviral agent, an antibacterial agent, an anti-inflammatory agent, and a scavenging agent.

4. The single-domain antibody construct of claim 1 , further comprising a diagnostic agent selected from the group consisting of: an imaging agent, a marker, a dye, a detectable moiety, and a label.

5. The single-domain antibody construct of claim 1 , further comprising a second single-domain antibody with a second binding domain, wherein the binding domain is configured to target lung cells; and the second binding domain is configured to treat or diagnose a disease or malignancy.

6. The single-domain antibody construct of claim 1 , further comprising a therapeutic agent with antiviral activity against a coronavirus.

7. The single-domain antibody construct of claim 1 , wherein the single-domain antibody further comprises:

a first framework region attached to an N-terminus of the first complementarity determining region;

a second framework region disposed between the first and second complementarity determining regions;

a third framework region disposed between the second and third complementarity determining regions; and

a fourth framework region attached to a C-terminus of the third complementarity determining region.

8. The single-domain antibody construct of claim 1 , wherein the first single-domain antibody is a polypeptide sequence having at least 99% sequence identity to any one of SEQ ID NOs: 76-100.

9. The single-domain antibody construct of claim 1 , wherein the first single-domain antibody comprises a first, second, third, and fourth framework region selected from the group consisting of the following combinations:

SEQ ID NOs.: 191, 221, 252, and 290, respectively; and

SEQ ID NOs: 190, 220, 251, and 290, respectively.

10. The single-domain antibody construct of claim 8 , wherein the first single-domain antibody is a polypeptide sequence corresponding to any one of SEQ ID NOs: 76-100.

11. The single-domain antibody construct of claim 1 , wherein the binding domain is configured to target mouse lung tissue.

12. The single-domain antibody construct of claim 11 , wherein the lung tissue comprises a cell selected from the group consisting of: airway cells in bronchial and bronchiolar epithelium, bronchial gland cells, alveolar cells, pulmonary vascular cells, and submucosal cells.

13. The single-domain antibody construct of claim 1 , wherein the single-domain antibody construct is expressed as a fusion protein coating protein gIIIp of M13 phage.

14. The single-domain antibody construct of claim 1 , wherein the first polypeptide comprising first, second, and third complementarity determining regions is selected from the group consisting of the following combinations:

SEQ ID NOs: 1, 26, and 51, respectively; and

SEQ ID NOs: 2, 27, and 52, respectively.

15. The single-domain antibody construct of claim 1 , wherein the first polypeptide comprising first, second, and third complementarity determining regions is selected from the group consisting of the following combinations:

SEQ ID NOs: 3, 28, and 53, respectively;

SEQ ID NOs: 4, 29, and 54, respectively;

SEQ ID NOs: 5, 30, and 55, respectively;

SEQ ID NOs: 6, 31, and 56, respectively;

SEQ ID NOs: 7, 32, and 57, respectively;

SEQ ID NOs: 8, 33, and 58, respectively; and

SEQ ID NOs: 9, 34, and 59, respectively.

16. The single-domain antibody construct of claim 1 , wherein the first polypeptide comprising first, second, and third complementarity determining regions is selected from the group consisting of the following combinations:

SEQ ID NOs: 11, 36, and 61, respectively;

SEQ ID NOs: 12, 37, and 62, respectively;

SEQ ID NOs: 13, 38, and 63, respectively;

SEQ ID NOs: 14, 39, and 64, respectively;

SEQ ID NOs: 15, 40, and 65, respectively;

SEQ ID NOs: 16, 41, and 66, respectively;

SEQ ID NOs: 17, 42, and 67, respectively;

SEQ ID NOs: 18, 43, and 68, respectively;

SEQ ID NOs: 19, 44, and 69, respectively;

SEQ ID NOs: 20, 45, and 70, respectively;

SEQ ID NOs: 21, 46, and 71, respectively;

SEQ ID NOs: 22, 47, and 72, respectively;

SEQ ID NOs: 23, 48, and 73, respectively;

SEQ ID NOs: 24, 49, and 74, respectively; and

SEQ ID NOs: 25, 50, and 75, respectively.

17. A vector comprising:

a phage configured to express a single-domain antibody with a binding domain, wherein the binding domain comprises:

a first polypeptide comprising first, second, and third complementarity determining regions selected from the group consisting of the following combinations:

SEQ ID NOs: 1, 26, and 51, respectively;

SEQ ID NOs: 2, 27, and 52, respectively;

SEQ ID NOs: 3, 28, and 53, respectively;

SEQ ID NOs: 4, 29, and 54, respectively;

SEQ ID NOs: 5, 30, and 55, respectively;

SEQ ID NOs: 6, 31, and 56, respectively;

SEQ ID NOs: 7, 32, and 57, respectively;

SEQ ID NOs: 8, 33, and 58, respectively;

SEQ ID NOs: 9, 34, and 59, respectively;

SEQ ID NOs: 10, 35, and 60, respectively;

SEQ ID NOs: 11, 36, and 61, respectively;

SEQ ID NOs: 12, 37, and 62, respectively;

SEQ ID NOs: 13, 38, and 63, respectively;

SEQ ID NOs: 14, 39, and 64, respectively;

SEQ ID NOs: 15, 40, and 65, respectively;

SEQ ID NOs: 16, 41, and 66, respectively;

SEQ ID NOs: 17, 42, and 67, respectively;

SEQ ID NOs: 18, 43, and 68, respectively;

SEQ ID NOs: 19, 44, and 69, respectively;

SEQ ID NOs: 20, 45, and 70, respectively;

SEQ ID NOs: 21, 46, and 71, respectively;

SEQ ID NOs: 22, 47, and 72, respectively;

SEQ ID NOs: 23, 48, and 73, respectively;

SEQ ID NOs: 24, 49, and 74, respectively; and

SEQ ID NOs: 25, 50, and 75, respectively.

18. The vector of claim 17 , wherein the phage comprises an M13 bacteriophage.

19. The vector of claim 17 , wherein the single-domain antibody is expressed as a fusion protein to coat protein gIIIp of M13 bacteriophage.

20. The vector of claim 17 , wherein the first polypeptide comprising first, second, and third complementarity determining regions is selected from the group consisting of the following combinations:

SEQ ID NOs: 1, 26, and 51, respectively; and

SEQ ID NOs: 2, 27, and 52, respectively.

Assignments (3)

References Cited (15)