Photo-controlled removal of targets in vitro and in vivo
This disclosure provides IR700-molecule conjugates and methods of their use to remove (e.g., separate or isolate) a target from a sample in vivo or from a subject in vitro. It is shown herein that exposure of IR700 to near infrared (NIR) light removes a portion of IR700, changing it from a hydrophilic molecule, to one that is hydrophobic, resulting in aggregation of IR700 and anything bound to it. For example, the disclosed IR700-molecule conjugates and methods provide photo-controlled ways to control the pharmacokinetics of a drug in vivo, and can be used to remove undesired agents from environmental or food samples or to isolate target molecules in a laboratory.
1. A method of removing a target from a sample, comprising:
contacting the sample with a phthalocyanine IRDye 700DX (IR700) conjugated to a molecule (IR700-molecule conjugate) to form an IR700-molecule conjugate-target complex, wherein the molecule of the IR700-molecule conjugate comprises a specific binding agent that specifically binds to the target;
irradiating the sample after the contacting step, at a wavelength of 660 to 710 nm and at a dose of at least 1 J cm −2 under conditions sufficient for cleaving and removing one or both of the hydrophilic side chains of the IR700 and generating a hydrophobic IR700-molecule conjugate-target complex;
incubating the sample, after the contacting step under conditions that allow the hydrophobic IR700-molecule conjugate-target complex to aggregate; and
separating the aggregate of the hydrophobic IR700-molecule conjugate-target complex from the sample, thereby removing the target from the sample, wherein the target is a protein, peptide, lectin, carbohydrate, metal, nucleic acid molecule, recreational drug, small organic molecule, pathogen, or spore.
2. The method of claim 1 , wherein the IR700-molecule conjugate comprises an IR700-antibody conjugate, IR700-antibody fragment conjugate, IR700-antibody mimetic molecule conjugate, IR700-hapten conjugate, IR700-lectin conjugate, IR700-protein conjugate, IR700-nucleic acid molecule conjugate, or IR700-functional nucleic acid conjugate,
wherein the antibody, antibody fragment, antibody mimetic molecule, hapten, lectin, protein, nucleic acid molecule, and functional nucleic acid specifically bind to the target molecule.
3. The method of claim 1 , wherein the sample is a food sample, environmental sample, reactor sample, fermentation sample, or sample obtained from a subject.
4. The method of claim 1 , wherein the sample is irradiated at a wavelength of 690 nm+/−20 nm or 690 nm+/−4 nm.
5. The method of claim 1 , wherein incubating the sample comprises centrifuging the sample to form a pellet.
6. The method of claim 1 , wherein incubating the sample comprises allowing the hydrophobic IR700-molecule conjugate-target complex to settle to the bottom of a container.
7. The method of claim 1 , wherein separating the hydrophobic IR700-molecule conjugate-target complex from the sample comprises collecting a supernatant after allowing the hydrophobic IR700-molecule conjugate-target complex to aggregate.
8. The method of claim 1 , further comprising measuring the target removed from the sample.
9. The method of claim 1 , wherein the sample is irradiated at a wavelength of 660 to 710 nm and at a dose of at least 4 J cm 2 .
10. The method of claim 1 , wherein the sample is irradiated at a wavelength of 660 to 710 nm and at a dose of at least 100 J cm 2 .
11. The method of claim 1 , wherein the method comprises irradiating the sample at a wavelength of 680 to 690 nm and at a dose of at least 4 J cm 2 .
12. A method of removing and detecting a target from a subject, comprising:
administering to the subject systemically, inhaltionally or intranasally, a therapeutically effective amount of a phthalocyanine IRDye 700DX (IR700) conjugated to a molecule (IR700-molecule), wherein the molecule comprises a specific binding agent that specifically binds to the target;
allowing the administered IR700-molecule to bind to the target via the specific binding agent;
irradiating the subject after the administering step, at a wavelength of 660 nm to 710 nm and at a dose of at least 4 J cm −2 under conditions sufficient for cleaving and removing one or both of the hydrophilic side chains of the IR700 and forming a hydrophobic IR700-molecule-target complex;
allowing the hydrophobic IR700-molecule-target complex to aggregate in the subject;
subsequently, after allowing to aggregate, obtaining from the subject a urine or bowel movement sample comprising the hydrophobic IR700-molecule-target complex aggregate; and
detecting in the urine or bowel movement sample the hydrophobic IR700-molecule-target complex aggregate, wherein detection of the aggregate indicates detection of the removed target from the subject.
13. The method of claim 12 , wherein the target is a protein, peptide, lectin, carbohydrate, metal, nucleic acid molecule, small organic molecule, recreational drug, pathogen, spore, or cell.
14. The method of claim 13 , wherein the target cell is a cell in a tumor.
15. The method of claim 14 , wherein the cell in the tumor is a tumor cell, immune cell, or cancer stem cell.
16. The method of claim 14 , wherein the target cell is a negative regulatory T-cell.
17. The method of claim 16 , wherein the negative regulatory T-cell is a CD4 + CD25 + FoxP3 + T cell, and the IR700-molecule conjugate comprises an anti-CD25 or anti-CLTA4 antibody.
18. The method of claim 13 , wherein the target cell is a lymphocyte, dendritic cell, or macrophage.
19. The method of claim 12 , wherein the IR700-molecule conjugate comprises an IR700-antibody conjugate, IR700-antibody fragment conjugate, IR700-antibody mimetic molecule conjugate, IR700-hapten conjugate, IR700-lectin conjugate, IR700-protein conjugate, IR700-nucleic acid molecule conjugate, or IR700-functional nucleic acid conjugate,
wherein the antibody, antibody fragment, antibody mimetic molecule, hapten, lectin, protein, nucleic acid molecule, and functional nucleic acid specifically bind to the target molecule.
20. The method of claim 13 , wherein the subject is irradiated at a wavelength of 690 nm+/−20 nm or 690 nm+/−4 nm.
21. The method of claim 12 , wherein irradiating the subject comprises using a device worn by the subject, wherein the device comprises a near infrared (NIR) light emitting diode (LED).
22. The method of claim 12 , wherein the subject has a cancer of the breast, liver, kidney, uterus, colon, ovary, prostate, pancreas, brain, cervix, bone, skin, or lung.