INDICATOR COMPOUNDS, DEVICES COMPRISING INDICATOR COMPOUNDS, AND METHODS OF MAKING AND USING THE SAME
Some embodiments disclosed herein pertain to indicator compounds used to detect the presence of and/or an amount of an analyte. In some embodiments, the indicator compounds are fusion proteins. In some embodiments, when the analyte binds to the indicator compound, the indicator compound undergoes a conformational change. In some embodiments, the conformational change results in a luminescent signal that allows quantification of the amount of analyte present.
1 . An indicator compound comprising:
a luminescent protein domain; and
an analyte binding protein domain;
wherein the indicator compound is configured to bind an analyte via the analyte binding protein domain;
wherein the indicator compound is configured to undergo a conformational change from a first conformation to a second conformation when the analyte is bound by the analyte binding protein domain;
wherein, when in the second conformation, the indicator compound is configured to receive an excitation wavelength and to emit a luminescent signal;
wherein a contiguous 100 amino acid portion of the analyte binding protein domain shares 80% to 100% identity, homology, or similarity to a contiguous 100 amino acid portion of one of SEQ ID NOs: 2, 15, 122-221, or 824-1746.
2 . (canceled)
3 . A method of manufacturing the indicator compound of claim 1 , comprising expressing the indicator compound encoded by a vector in a recombinant host cell, and isolating the indicator compound from the recombinant host cell.
4 . The method of claim 3 , wherein the vector comprises:
a nucleic acid sequence encoding the luminescent protein domain and/or one or more luminescing fragments of a luminescent protein; and
a nucleic acid sequence encoding the analyte binding protein domain.
5 . The method of claim 4 , wherein the vector further comprises a signal sequence.
6 . The indicator compound of claim 1 , wherein the luminescent protein domain shares 80% to 100% identity, homology, or similarity to one or more of SEQ ID NOS: 3-13 and 16-20.
7 . The indicator compound of claim 1 , wherein the indicator compound is encoded by DNA that shares 80% to 100% identity, homology, or similarity to one or more of SEQ ID NOS: 21-33.
8 . The indicator compound of claim 1 , a contiguous 100 amino acid portion of the luminescent protein domain shares 80% to 100% identity, homology, or similarity to a contiguous 100 amino acid portion of one of SEQ ID NOs: 1, 14, 34-121, or 222-823.
9 . The indicator compound of claim 1 , wherein the luminescent protein domain is a split luminescent protein.
10 . The indicator compound of claim 1 , wherein the luminescent protein domain is a fluorescent protein domain.
11 . The indicator compound of claim 1 , wherein the analyte binding protein domain shares 80% to 100% identity, homology, or similarity to one of SEQ ID NOs: 2, 15, 122-221, or 824-1746.
12 . The indicator compound of claim 1 , wherein the analyte binding protein domain is a periplasmic protein sugar binding domain.
13 . The indicator compound of claim 1 , wherein the luminescent protein domain shares 80% to 100% identity, homology, or similarity to one or more of SEQ ID NOs: 1, 14, 34-121, or 222-823.
14 . A method for detecting glucose in a patient comprising:
administering an indicator compound to the patient, the indicator compound comprising:
a luminescent protein domain; and
a glucose binding protein domain;
wherein the indicator compound is configured to bind glucose via the glucose binding protein domain;
wherein the indicator compound is configured to undergo a conformational change from a first conformation to a second conformation when glucose is bound by the glucose binding protein domain;
wherein, when in the second conformation, the indicator compound is configured to receive an excitation wavelength and to emit a luminescent signal;
wherein a contiguous 100 amino acid portion of the glucose binding protein domain shares 80% to 100% identity, homology, or similarity to a contiguous 100 amino acid portion of one of SEQ ID NOs: 2, 15, 122-221, or 824-1746.
15 . The method of claim 14 , further comprising quantifying a glucose level in the patient based on a change in luminescence of the indicator compound.
16 . The method of claim 14 , wherein the indicator compound is administered to the patient using a cell.
17 . The method of claim 16 , wherein the cell is autologous to the patient.
18 . The method of claim 16 , wherein the cell comprises DNA having a region that encodes the indicator compound.
19 . The method of claim 16 , wherein the cell is part of a skin graft received by the patient.
20 . The method of claim 16 , wherein the cell is a keratinocyte, melanocyte, Langerhans cell, Merkel cell, immune cell, macrophage, epithelial cell, or red blood cell.
21 . The method of claim 14 , further comprising illuminating the indicator compound with light having a first maximum wavelength and detecting an emission of light at a second maximum wavelength.