Detecting targets in heterologous mixtures
First Claim
1. A method, comprising:
- a) providing a conjugate comprising a detectable moiety linked to a photoreactive moiety, wherein said detectable moiety comprises a biotinyl moiety, wherein said photoreactive moiety comprises at least one group covalently bound to a substrate, wherein said substrate is selected from proteins, peptides, amino acids, lipids, cells, virus particles, fatty acids, nucleic acids, nucleotides, nucleosides, polysaccharides and inorganic molecules; and
b) treating said conjugate with electromagnetic radiation to release said detectable moiety.
1 Assignment
0 Petitions
Accused Products
Abstract
This invention relates to agents and conjugates that can be used to detect and isolate target components from complex mixtures such as nucleic acids from biological samples, cells from bodily fluids, and nascent proteins from translation reactions. Agents comprise a detectable moiety bound to a photoreactive moiety. Conjugates comprise agents coupled to substrates by covalent bounds which can be selectively cleaved with the administration of electromagnetic radiation. Targets substances labeled with detectable molecules can be easily identified and separated from a heterologous mixture of substances. Exposure of the conjugate to radiation releases the target in a functional form and completely unaltered. Using photocleavable molecular precursors as the conjugates, label can be incorporated into macromolecules, the nascent macromolecules isolated and the label completely removed. The invention also relates to targets isolated with these conjugates which may be useful as pharmaceutical agents or compositions that can be administered to humans and other mammals. Useful compositions include biological agents such as nucleic acids, proteins, lipids and cytokines. Conjugates can also be used to monitor the pathway and half-life of pharmaceutical composition in vivo and for diagnostic, therapeutic and prophylactic purposes. The invention also relates to kits comprised of agents and conjugates that can be used for the detection of diseases, disorders and nearly any individual substance in a complex background of substances.
-
Citations
24 Claims
-
1. A method, comprising:
-
a) providing a conjugate comprising a detectable moiety linked to a photoreactive moiety, wherein said detectable moiety comprises a biotinyl moiety, wherein said photoreactive moiety comprises at least one group covalently bound to a substrate, wherein said substrate is selected from proteins, peptides, amino acids, lipids, cells, virus particles, fatty acids, nucleic acids, nucleotides, nucleosides, polysaccharides and inorganic molecules; and
b) treating said conjugate with electromagnetic radiation to release said detectable moiety. - View Dependent Claims (2, 3, 4, 5, 6)
-
-
7. A method, comprising:
-
a) providing i) a heterologous mixture comprising a target molecule, and ii) a conjugate comprising a detectable moiety linked to a photoreactive moiety, wherein said detectable moiety comprises a biotinyl moiety, wherein said photoreactive moiety comprises at least one group covalently bound to a substrate, wherein said substrate is selected from proteins, peptides, amino acids, lipids, cells, virus particles, fatty acids, nucleic acids, nucleotides, nucleosides, polysaccharides and inorganic molecules;
b) contacting said conjugate with said heterologous mixture under conditions such that said substrate binds to said target molecule to form a coupled conjugate; and
c) treating said coupled conjugate with electromagnetic radiation to release said detectable moiety. - View Dependent Claims (8, 9, 10, 11, 12)
-
-
13. A method, comprising:
-
a) providing i) a sample comprising a plurality of target molecules, and ii) a conjugate preparation, said conjugate comprising a detectable moiety linked to a photoreactive moiety, wherein said detectable moiety comprises a biotinyl moiety, wherein said photoreactive moiety comprises at least one group covalently bound to a substrate, wherein said substrate is selected from proteins, peptides, amino acids, nucleic acids, nucleotides, and nucleosides;
b) contacting said sample with a first portion of said conjugate preparation under conditions such that said substrate binds to at least one of said plurality of target molecules to form a first coupled conjugate;
c) treating said first coupled conjugate with electromagnetic radiation to release said detectable moiety;
d) contacting said sample with a second portion of said conjugate preparation under conditions such that said substrate binds to at least one of said plurality of target molecules to form a second coupled conjugate; and
e) treating said second coupled conjugate with electromagnetic radiation to release said detectable moiety. - View Dependent Claims (14, 15, 16, 17, 18, 22, 23, 24)
-
-
19. A method, comprising:
-
a) providing i) a sample comprising a plurality of target molecules, and ii) a conjugate preparation, said conjugate comprising a detectable moiety linked to a photoreactive moiety, wherein said detectable moiety comprises a fluorescent moiety, wherein said photoreactive moiety comprises at least one group covalently bound to a substrate, wherein said substrate is selected from proteins, peptides, amino acids, nucleic acids, nucleotides, and nucleosides;
b) contacting said sample with a first portion of said conjugate preparation under conditions such that said substrate binds to at least one of said plurality of target molecules to form a first coupled conjugate;
c) treating said first coupled conjugate with electromagnetic radiation to release said detectable moiety;
d) contacting said sample with a second portion of said conjugate preparation under conditions such that said substrate binds to at least one of said plurality of target molecules to form a second coupled conjugate; and
e) treating said second coupled conjugate with electromagnetic radiation to release said detectable moiety. - View Dependent Claims (20, 21)
-
Specification