Perturbation beads for use in assays
Disclosed are uniquely labeled perturbation beads used alone or in combination with a capture bead. These perturbation beads are useful in drug discovery especially in assays conducted in a mapped assay device. Such a uniquely labeled perturbation bead, when used in an examination area of a mapped assay device, allows for correlating the sequence of nucleic acids and other components from a lysed cell to the specific examination area where the nucleic acid was retrieved.
1 . An optically labeled perturbation microbead from a combinatorial library of microbeads, wherein said optically labeled perturbation microbead comprises: a structure of Formula I′;
wherein:
PB is a perturbation microbead;
L and L 1 are each a releasable linker, wherein L and L 1 can be the same or different;
PE is a perturbation element comprising a chemical compound, wherein each chemical compound comprises a plurality of chemical building blocks, wherein PE is releasably bound to the perturbation microbead via the releasable linker L 1 ;
W is a set of optically detectable labels that uniquely identifies the perturbation microbead, wherein each optically detectable label or each set of optically detectable labels:
(i) is optically distinguishable from all other optically detectable labels of the set of optically detectable labels; and
(ii) uniquely corresponds to a chemical building block of the plurality of chemical building blocks;
Q is a perturbation oligonucleotide that codes for the chemical compound, wherein each perturbation oligonucleotide (Q) comprises a sequence of oligonucleotide strands, wherein each oligonucleotide strand of the sequence of oligonucleotide strands uniquely codes for both:
i) a chemical building block of the plurality of chemical building blocks in each perturbation element (PE); and
(ii) an optically detectable label corresponding to the chemical building block of the plurality of chemical building blocks in each perturbation element (PE);
Q 1 is an oligonucleotide binding element that is:
(i) complementary to a complementary capture binding element on a capture bead; or
(ii) complementary to a cellular component released from a lysed cell;
m is an integer from 1 to 100, wherein m denotes a number of the sets of optically detectable labels (W) bound to the perturbation microbead (PB);
n is a number of (L-Q-Q 1 ) groups bound to the perturbation microbead (PB), wherein n is in a range from 1×10 5 to 6.02×10 17 ; and
p is a number of (L 1 -PE) groups that are bound to the perturbation microbead (PB), wherein p is in a range from 1×10 5 to 6.02×10 17 .
2 . The optically labeled perturbation microbead of claim 1 , wherein the oligonucleotide binding element (Q 1 ) comprises a poly-T oligonucleotide.
3 . The optically labeled perturbation microbead of claim 1 , wherein the releasable linkers L 1 are independent from the releasable linkers L.
4 . The optically labeled perturbation microbead of claim 1 , wherein the releasable linkers L 1 are different types of linkers from the releasable linkers L.
5 . The optically labeled perturbation microbead of claim 1 , wherein releasable linker L and releasable linker L 1 are different, such that releasable linker L is configured to be cleaved by a first stimulus and the releasable linker L 1 is configured to be cleaved by a second stimulus that is different from the first stimulus.
6 . The optically labeled perturbation microbead of claim 1 , wherein the set of optically detectable labels (W) comprises a set of quantum dots, wherein each quantum dot of the set of quantum dots is configured to generate light of a different wavelength from a remainder of the set of quantum dots.
7 . The optically labeled perturbation microbead of claim 1 , wherein the set of optically detectable labels (W) comprises a set of micro-components, wherein each micro-component of the set of micro-components is embedded in the perturbation microbead.
8 . The optically labeled perturbation microbead of claim 1 , wherein each optically detectable label of the set of optically detectable labels (W) is bound to the perturbation microbead (PB) through one or more of an amide bond or an electrostatic attachment.
9 . The optically labeled perturbation microbead of claim 1 , wherein the set of optically detectable labels (W) comprises optically distinguishable micro-components selected from the group consisting of a set of optically detectable semi-conductor microchips, a set of optically detectable silicon oxide-coated micropucks, and a combination thereof.
10 . The optically labeled perturbation microbead of claim 1 , wherein the perturbation microbead (PB) is a polystyrene resin bead configured for use in combinatorial library synthesis.
11 . The optically labeled perturbation microbead of claim 9 , wherein the set of optically detectable labels (W) is attached to the perturbation microbead (PB) and comprises optically detectable labels having a random distribution of different sizes and shapes.