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US 2026/0169002 A1Published application
Profiling newly-synthesized glycoproteins as molecular signatures of injury in donor organs
AssigneesCarnegie Mellon University, University of Pittsburgh—Of the Commonwealth System of Higher Education
InventorsXi Ren, Zihan Ling, Kentaro Noda, Pablo Sanchez
Life of this patent
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Abstract
Methods and materials for improving outcomes for ex vivo perfused organs are provided herein. For example, this document provides methods and materials that can be used to assess and treat lungs during ex vivo perfusion prior to transplantation.
Claims17
A method for assessing an organ or a portion thereof removed from a mammal for transplant suitability, said method comprising:
- (a) perfusing said organ or said portion thereof with an aqueous solution comprising (i) an azide-containing chemical probe, thereby azide-tagging glycosylated polypeptides synthesized within cells of said organ or said portion thereof during said perfusing, or (ii) an alkyne-containing chemical probe, thereby alkyne-tagging glycosylated polypeptides synthesized within cells of said organ or said portion thereof during said perfusing;
- (b) obtaining a sample of said organ after said perfusing, and generating an extract containing said azide- or alkyne-tagged glycosylated polypeptides;
- (c) contacting said azide- or alkyne-tagged glycosylated polypeptides in said extract with desthiobiotin, thereby generating desthiobiotinylated glycosylated polypeptides, wherein said desthiobiotin is alkyne-desthiobiotin when said glycosylated polypeptides are azide-tagged, and wherein said desthiobiotin is azide-desthiobiotin when said glycosylated polypeptides are alkyne-tagged;
- (d) contacting said desthiobiotinylated glycosylated polypeptides with streptavidin coupled to a solid substrate, thereby coupling said desthiobiotinylated glycosylated polypeptides to said solid substrate;
- (e) washing said solid substrate;
- (f) eluting said desthiobiotinylated glycosylated polypeptides from said solid substrate;
- (g) measuring, within said eluate, the levels of one or more desthiobiotinylated glycoprotein markers of graft injury;
- and
- (h) determining that said organ is suitable for transplant when said eluate does not contain altered levels of said one or more desthiobiotinylated glycoprotein markers of said graft injury, as compared to a control eluate from a corresponding organ without said graft injury, or determining that said organ may not be suitable for transplant when said eluate contains altered levels of said one or more desthiobiotinylated glycoprotein markers of said graft injury, as compared to a control eluate from a corresponding organ without said graft injury.
6 more elements
A method for assessing and treating an organ removed from a mammal, said method comprising:
- (a) perfusing said organ or said portion thereof with an aqueous solution comprising (i) an azide-containing chemical probe, thereby azide-tagging glycosylated polypeptides synthesized within cells of said organ or said portion thereof during said perfusing, or (ii) an alkyne-containing chemical probe, thereby alkyne-tagging glycosylated polypeptides synthesized within cells of said organ or said portion thereof during said perfusing;
- (b) obtaining a sample of said organ after said perfusing, and generating an extract containing said azide- or alkyne-tagged glycosylated polypeptides;
- (c) contacting said azide- or alkyne-tagged glycosylated polypeptides in said extract with desthiobiotin, thereby generating desthiobiotinylated glycosylated polypeptides, wherein said desthiobiotin is alkyne-desthiobiotin when said glycosylated polypeptides are azide-tagged, and wherein said desthiobiotin is azide-desthiobiotin when said glycosylated polypeptides are alkyne-tagged;
- (d) contacting said desthiobiotinylated glycosylated polypeptides with streptavidin coupled to a solid substrate, thereby coupling said desthiobiotinylated glycosylated polypeptides to said solid substrate;
- (e) washing said solid substrate;
- (f) eluting said desthiobiotinylated glycosylated polypeptides from said solid substrate;
- (g) measuring, within said eluate, the levels of one or more desthiobiotinylated glycoprotein markers of warm ischemia injury;
- (h) determining that said eluate contains altered levels of said one or more desthiobiotinylated glycoprotein markers of graft injury, as compared to a control eluate from a corresponding organ without graft injury;
- and
- (i) treating said organ with a calcineurin pathway inhibitor.
7 more elements
Dependencies are inferred from the claim text.