Multiplex analysis using membrane-bound sensitizers

• US 7,217,531 B2
• Filed: 08/27/2005
• Issued: 05/15/2007
• Est. Priority Date: 03/05/2002
• Status: Expired due to Fees

• Current Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Original Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Inventors: Singh, Sharat, Chan-Hui, Po-Ying, Kirakossian, Hrair
• Primary Examiner(s): Tate; Christopher R.
• Assistant Examiner(s): Davis; Deborah A.
• Application Number:
  US11/214,234
• Publication Number:
  US 20060024846A1
• Time in Patent Office: 626 Days
• Field of Search: 436/8, 436/518, 435/7.1, 435/7.2, 435/7.8
• US Class Current: 435/7.1
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/567 utilising isolate of tissue or organ as binding agent
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing

Abstract

The present invention is directed to methods and compositions for determining the presence, absence, and/or amounts of one or more membrane-associated analytes in a sample. In accordance with the invention, binding compounds derivatized with releasable molecular tags specifically bind to selected membrane-associated analytes, after which the molecular tags are released upon activation of cleavage moieties, or sensitizers, anchored in the same membrane as the membrane-associated analytes. The released molecular tags are then identified by their distinct separation and detection characteristics.

Claims

1. A composition for determining the presence or absence of one or more membrane-associated analytes, the composition comprising:

a sensitizer-treated membrane containing one or more membrane associated analytes, the sensitizer-treated membrane comprising a cell surface membrane having photosensitizers attached, the photosensitizers each having a lipophilic moiety for anchoring the photosensitizers in the cell surface membrane; and

one or more binding compounds, such that there is at least one binding compound specific for each of the one or more membrane-associated analytes, each binding compound having one or more molecular tags, each molecular tag being attached by a cleavable linkage, and the molecular tags of each binding compound being distinguishable from those of every other binding compound by one or more physical and/or optical characteristics;

wherein at least one binding compound of the plurality is specifically bound to at least one membrane-associated analyte.

2. The composition of claim 1 wherein said one or more membrane-associated analytes are selected from the group consisting of G-protein coupled receptors “

GPCRs.”

3. The composition of claim 1 wherein said one or more binding compounds each comprises an antibody composition.

4. The composition of claim 3 wherein said antibody composition is a monoclonal antibody or a polyclonal antibody composition.

5. The composition according to claims 1, 2, 3, or 4 wherein said one or more binding compounds is a plurality of said binding compounds in the range of from 2 to 50 and wherein said one or more physical characteristics is electrophoretic mobility.

6. The composition of claim 5 wherein said molecular tags upon cleavage from different said binding compounds form distinct peaks in an electropherogram.