IP Library › Granted Patent US 12,241,839
Granted Patent B2
US 12,241,839 · App. 17/105,925 · Granted Mar 4, 2025

Multipartite luciferase peptides and polypeptides

Inventors: Virginia Kincaid (Madison, WI); Lance P. Encell (Madison, WI); Mary Hall (Madison, WI); Michael Killoran (Madison, WI); Keith Wood (Madison, WI); Melanie Dart (Madison, WI); Trish Hoang (Madison, WI)
Assignee: Promega Corporation
G01N21/763C12N9/0069G01N33/582
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Quick Facts
Patent No.
US 12,241,839
App. No.
17/105,925
Granted
Mar 4, 2025
Kind
B2
Abstract

Provided herein are bioluminescent polypeptides and compositions and methods for the assembly of a tripartite or multipartite bioluminescent complex. In particular embodiments, a bioluminescent complex is formed upon the interaction of three or more peptide and/or polypeptide components.

Claims (18)

1. A system or kit comprising two or more peptide and/or polypeptide components collectively comprising 100% sequence identity to SEQ ID NO: 788 or SEQ ID NO: 789; wherein the two or more peptide and/or polypeptide components are capable of forming a bioluminescent complex upon interacting with each other; wherein a bioluminescent signal produced by the bioluminescent complex in the presence of a coelenterazine or a coelenterazine analog substrate is substantially increased when compared to a bioluminescent signal produced by one of the peptide and/or polypeptide components individually in the presence of the coelenterazine substrate; wherein the system or kit comprises a polypeptide that corresponds structurally to 8 or fewer β strands of a Oplophorus gracilirostris luciferase.

2. The system or kit of claim 1 , comprising a polypeptide component having 100% sequence identity to SEQ ID NO: 790 and one or more complementary peptides collectively having 100% sequence identity to SEQ ID NO: 794.

3. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 791 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 795.

4. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 792 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 796.

5. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 793 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 797.

6. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 790 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 798.

7. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 791 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 799.

8. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 792 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 800.

9. The system or kit of claim 1 , wherein the polypeptide comprises 100% sequence identity to SEQ ID NO: 793 and the one or more complementary peptides collectively comprise 100% sequence identity to SEQ ID NO: 801.

10. The system or kit of claim 1 , wherein one or more of the polypeptide and/or peptide components are present as fusions with one or more additional amino acid sequences.

11. The system or kit of claim 10 , wherein the additional amino acid sequence is selected from the group consisting of a protein of interest, an interaction element, a co-localization element, and a binding moiety.

12. The system or kit of claim 10 , wherein the additional amino acid sequence is a binding moiety selected from the group consisting of antibody (polyclonal, monoclonal, and/or recombinant), antibody fragment, protein A, an Ig binding domain of protein A, protein G, an Ig binding domain of protein G, protein A/G, an Ig binding domain of protein A/G, protein L, an Ig binding domain of protein L, protein M, an Ig binding domain of protein M, oligonucleotide probe, peptide nucleic acid, DARPin, aptamer, affimer, a purified protein (either the analyte itself or a protein that binds to the analyte), and analyte binding domain(s) of proteins.

13. The system or kit of claim 10 , wherein the additional amino acid sequence is a first interaction polypeptide that is configured to form a complex with a second interaction polypeptide upon contact of the first interaction polypeptide and the second interaction polypeptide.

14. The system or kit of claim 10 , wherein the additional amino acid sequence is a first co-localization polypeptide that is configured to co-localize within a cellular compartment, a cell, a tissue, or an organism within a with a second co-localization polypeptide.

15. The system or kit of claim 10 , wherein the additional amino acid sequence is a protein of interest and is a candidate drug target.

16. A bioluminescent complex comprising the two or more peptide and/or polypeptide components of the system or kit of claim 1 .

17. A method comprising contacting the bioluminescent complex of claim 16 with a substrate for the bioluminescent complex.

18. The method of claim 17 wherein the substrate is coelenterazine or furimazine.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 13, 2022
From: DART, MELANIE; HOANG, TRISH
To: PROMEGA CORPORATION
Reel/Frame 061071/0178 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 14, 2021
From: KINCAID, VIRGINIA; ENCELL, LANCE P; HALL, MARY; KILLORAN, MICHAEL; WOOD, KEITH
To: PROMEGA CORPORATION
Reel/Frame 056247/0943 →
Continuity (2)
Provisional Application 62941255 · Nov 27, 2019
Related Publication 20210262941A1 · Aug 26, 2021
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