Multiplex Digital Immuno-Sensing Using a Library of Photocleavable Mass Tags
First Claim
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1. A method for detecting the presence of an agent in a sample comprising:
- (a) contacting the sample with a solid substrate having affixed thereto a first antibody which binds to the agent, wherein the contacting is performed under conditions which would permit the first antibody to bind to the agent if present in the sample;
(b) removing any unbound sample from the solid substrate;
(c) contacting the solid substrate with a second antibody which binds to the agent concurrently with the first antibody, wherein the second antibody has a mass tag cleavably affixed thereto, and wherein the contacting is performed under conditions which would permit the second antibody to bind to the agent if present in the sample;
(d) removing any unbound second antibody;
(e) cleaving the mass tag from any bound second antibody; and
(f) detecting the presence of any cleaved mass tag,wherein the presence of cleaved mass tag indicates that the agent is present in the sample.
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Abstract
This invention provides methods, compositions and kits for immunosensing using photocleavable mass tags.
137 Citations
47 Claims
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1. A method for detecting the presence of an agent in a sample comprising:
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(a) contacting the sample with a solid substrate having affixed thereto a first antibody which binds to the agent, wherein the contacting is performed under conditions which would permit the first antibody to bind to the agent if present in the sample; (b) removing any unbound sample from the solid substrate; (c) contacting the solid substrate with a second antibody which binds to the agent concurrently with the first antibody, wherein the second antibody has a mass tag cleavably affixed thereto, and wherein the contacting is performed under conditions which would permit the second antibody to bind to the agent if present in the sample; (d) removing any unbound second antibody; (e) cleaving the mass tag from any bound second antibody; and (f) detecting the presence of any cleaved mass tag, wherein the presence of cleaved mass tag indicates that the agent is present in the sample. - View Dependent Claims (5, 6, 7, 8, 9, 10, 14, 16, 19)
wherein X is H, F, OMe, or (OMe)2
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2. (canceled)
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3. A method for detecting the presence of an agent in a sample comprising:
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(a) contacting the sample with a solid substrate which binds to the agent, wherein the contacting is performed under conditions which would permit the solid substrate to bind to the agent if present in the sample; (b) removing any unbound sample from the solid substrate; (c) contacting the solid substrate with an antibody which binds to the agent concurrently with the solid substrate, wherein the antibody has a mass tag cleavably affixed thereto, and wherein the contacting is performed under conditions which would permit the antibody to bind to the agent if present in the sample; (d) removing any unbound antibody; (e) cleaving the mass tag from any bound antibody; and (f) detecting the presence of any cleaved mass tag, wherein the presence of cleaved mass tag indicates that the agent is present in the sample. - View Dependent Claims (41, 42, 43, 44, 45, 46, 47)
wherein X is H, F, OMe, or (OMe)2.
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4. (canceled)
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11-13. -13. (canceled)
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15. (canceled)
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17-18. -18. (canceled)
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20. (canceled)
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21. A composition of matter comprising:
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(a) a solid substrate; (b) a first antibody bound to the solid substrate, wherein the first antibody recognizes an agent; (c) the agent recognized by the first antibody, wherein the agent is bound to the first antibody; and (d) a second antibody which recognizes the agent bound concurrently to the first antibody, wherein the second antibody is bound to the agent, and wherein the second antibody has a mass tag cleavably affixed thereto.
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22-40. -40. (canceled)
Specification