Non-invasive localization of a light-emitting conjugate in a mammal

• US 5,650,135 A
• Filed: 07/01/1994
• Issued: 07/22/1997
• Est. Priority Date: 07/01/1994
• Status: Expired due to Term

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Contag, Christopher H., Contag, Pamela R., Benaron, David A.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): GRASER, JENNIFER E
• Application Number:
  US08/270,631
• Time in Patent Office: 1,117 Days
• Field of Search: 424/9, 424/9.1, 424/193.1, 424/258.1, 424/93.2, 424/9.6, 424/9.61, 436/800, 536/25.32, 435/7.35, 435/7.9, 435/968, 435/172.3, 435/8
• US Class Current: 424/9.1
• CPC Class Codes: A61K 49/0013 Luminescence
• CPC Class Codes: A61K 49/0084 liposome, i.e. bilayered vesicular structure
• CPC Class Codes: C07K 14/195 from bacteria
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: C12Q 1/66 involving luciferase
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine

Abstract

Methods and compositions for detecting and localizing light originating from a mammal are disclosed. Also disclosed are methods for targeting light emission to selected regions, as well as for tracking entities within the mammal. In addition, animal models for disease states are disclosed, as are methods for localizing and tracking the progression of disease or a pathogen within the animal, and for screening putative therapeutic compounds effective to inhibit the disease or pathogen.

Claims

1. A noninvasive method for detecting the localization of an entity under study from within a mammalian subject, comprising

(a) administering to the subject a conjugate or transformed cell of the entity and a light-generating moiety or a transformed cell expressing the light-generating moiety,

(b) after a period of time in which the conjugate or transformed cell can achieve localization in the subject, immobilizing the subject within the detection field of a photodetector device,

(c) maintaining the subject in an immobilized condition,

(d) during said maintaining, measuring photon emission from the light-generating moiety, localized in the subject, with the photodetector device until an image of photon emission can be constructed, and

(e) detecting said image through an opaque tissue of said mammal.

2. The method of claim 1, which further includes repeating steps (b) through (e) at selected intervals,

wherein said repeating is effective to track the localization of the entity in the subject over time.

3. The method of claim 1, where said measuring is carried out with an intensified charge-coupled photodetector device.

4. The method of claim 1, for detecting the localization of infection by a pathogen in a mammalian subject, where said administering includes administering a conjugate or transformed cell that contains an infection-targeting moiety.

5. The method of claim 1, where said administering includes administering a conjugate that is a particle containing a light-generating moiety.

6. The method of claim 1, for detecting the localization of infection by a pathogen in an animal model, where the entity under study is the pathogen.

7. The method of claim 6, where the pathogen is Salmonella.

8. The method of claim 1, where the light-generating moiety is a light-generating protein.

9. The method of claim 8, where the protein is selected from the group consisting of luciferase, yellow fluorescent protein and ferredoxin IV.

10. The method of claim 8, where the entity under study is a transformed cell, and the light-generating moiety is a product of a heterologous gene expressed by the cell.

11. The method of claim 10, where expression of the heterologous gene is under the control of an activatable promoter.

13. The method of claim 1, wherein step (d) involves measuring changes in the level of photon emission over time of said light-generating source.

14. The method of claim 1, wherein at least one light-generating source has the capability to emit light of a wavelength in the range of between 300 nm and 1100 nm.

15. The method of claim 14, wherein at least one light-generating source has the capability to emit light of a wavelength of 550 nm.

16. The method of claim 1, wherein said photodetector device comprises one or a plurality of photodetector device elements.

17. The method of claim 16, wherein one photodetector device element is a charge-coupled device (CCD) camera.

18. The method of claim 1, wherein said step (d) involves determining a measurement that is a function of a local condition of the light-generating moiety.

19. The method of claim 1, wherein said image in step (e) reflects a state of a condition selected from the group consisting of oxygen levels, determination of presence of tumor cells, localization of tumor cells, determination of presence of an inflammation, localization of an inflammation, determination of presence of an infection, and localization of an infection.

20. The method of claim 1, wherein said image in step (e) reflects the oxygen level.

12. A noninvasive method for detecting the level of an entity under study in a mammalian subject over time, comprising

(a) administering to the subject a conjugate of the entity and a light-generating moiety or a transformed cell expressing the light-generating moiety,

(b) placing the subject within the detection field of a photodetector device,

(c) maintaining the subject in the detection field of the device,

(d) during said maintaining, measuring photon emission from the light-generating moiety, in the subject, with the photodetector device, and

(e) repeating steps (b) through (d) at selected intervals, wherein said repeating is effective to detect changes in the level of the entity in the subject over time.