In vivo detection of phospholipase activation
AssigneeThe Trustees of the University of Pennsylvania
InventorsEdward J. Delikatny, Anatoliy V. Popov, Gang Zheng, Theresa Mawn
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Abstract
The present invention is directed to a phospholipid-based NIR molecular beacon, having a phospholipid moiety; with an NIR fluorophore moiety covalently linked to a phospholipid glycerol backbone and a quencher moiety covalently linked to the phospholipid glycerol backbone. Additionally, provided herein is methods of analyzing a sample for the presence of a phospholipase and methods of identifying the activity of a phospholipase in vivo utilizing phospholipid-based NIR molecular beacon.
Claims10
A method of analyzing a sample for the presence of a phospholipase, the method comprising:
- contacting a sample suspected of comprising a phospholipase with a phospholipid-based near infrared (NIF) molecular beacon comprising (i) a phospholipid moiety with a glycerol backbone;
- (ii) a first fluorophore moiety covalently linked to the glycerol backbone either directly or via a linker, wherein said first fluorophore moiety is an NIR fluorophore moiety;
- and (iii) a fluorescence quencher moiety covalently linked to said phospholipid glycerol backbone either directly or via a linker, wherein the fluorophore moiety, the fluorescence quencher moiety or both are uncharged when covalently linked at either or both sn-1 or sn-2 fatty acyl positions;
- and
- detecting the NIR fluorescence of said NIR fluorophore moiety as a function of time, wherein an increase in the NIR fluorescence as a function of time correlates with the presence of the phospholipase in the sample.
A method of monitoring the activity of a phospholipase in vivo, comprising the steps of:
- exposing a tissue to a phospholipid-based near infrared (NIR) molecular beacon under conditions effective to permit said phospholipase to cleave said beacon, wherein said beacon comprises (i) a phospholipid moiety with a glycerol backbone;
- (ii) a first fluorophore moiety covalently linked to the glycerol backbone either directly or via a linker, wherein said first fluorophore moiety is an NIR fluorophore moiety;
- and (iii) a fluorescence quencher moiety covalently linked to said phospholipid glycerol backbone either directly or via a linker, wherein the fluorophore moiety, the fluorescence quencher moiety or both are uncharged when covalently linked at either or both sn-1 or sn-2 fatty acyl positions;
- and
- detecting the fluorescence of said fluorophore moiety as a function of time, wherein an increase in the quantity or rate of accumulation of fluorescence as compared to a control reaction identifies the activity of the phospholipase.
Dependencies are inferred from the claim text.