Multiplex analysis using membrane-bound sensitizers
View Patent ↗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.
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.