Methods for conducting multiplexed assays
The invention relates to methods for conducting solid-phase binding assays. One example is an assay method having improved analyte specificity where specificity is limited by the presence of non-specific binding interactions.
1. A pair of complementary oligonucleotides, wherein each oligonucleotide has a length of 10 to 50 bases, wherein each of the oligonucleotides is modified with a linking agent, and wherein each of the pair of complementary oligonucleotides comprises a sequence selected from:
Pair
Sequence
1
acatcggtagtt (SEQ ID NO: 1)
aactaccgatgt (SEQ ID NO: 2)
3
agaagaagatcc (SEQ ID NO: 5)
ggatcttcttct (SEQ ID NO: 6)
6
atcattaccacc (SEQ ID NO: 11)
ggtggtaatgat (SEQ ID NO: 12)
7
attaacgggagc (SEQ ID NO: 13)
gctcccgttaat (SEQ ID NO: 14)
8
cagaggtcttaa (SEQ ID NO: 15)
ttaagacctctg (SEQ ID NO: 16)
9
caggtgtccatt (SEQ ID NO: 17)
aatggacacctg (SEQ ID NO: 18)
11
cctacgatatac (SEQ ID NO: 21)
gtatatcgtagg (SEQ ID NO: 22)
12
cgaatgtagagt (SEQ ID NO: 23)
actctacattcg (SEQ ID NO: 24)
13
cggtttgagata (SEQ ID NO: 25)
tatctcaaaccg (SEQ ID NO: 26)
16
gacataaagcga (SEQ ID NO: 31)
tcgctttatgtc (SEQ ID NO: 32)
17
gccatagtctct (SEQ ID NO: 33)
agagactatggc (SEQ ID NO: 34)
18
gctaattcacca (SEQ ID NO: 35)
tggtgaattagc (SEQ ID NO: 36)
20
gttgattctgtc (SEQ ID NO: 39)
gacagaatcaac (SEQ ID NO: 40)
23
ttccacttaggg (SEQ ID NO: 45)
ccctaagtggaa (SEQ ID NO: 46)
25
tttcccttgcta (SEQ ID NO: 49)
tagcaagggaaa (SEQ ID NO: 50).
2. A kit comprising at least two pairs of complementary oligonucleotides, wherein each oligonucleotide has a length of 10 to 50 bases, wherein each of the oligonucleotides is modified with a linking agent, and wherein the pairs of complementary oligonucleotides comprise a sequence selected from:
Pair
Sequence
1
acatcggtagtt (SEQ ID NO: 1)
aactaccgatgt (SEQ ID NO: 2)
3
agaagaagatcc (SEQ ID NO: 5)
ggatcttcttct (SEQ ID NO: 6)
6
atcattaccacc (SEQ ID NO: 11)
ggtggtaatgat (SEQ ID NO: 12)
7
attaacgggagc (SEQ ID NO: 13)
gctcccgttaat(SEQ ID NO: 14)
8
cagaggtcttaa (SEQ ID NO: 15)
ttaagacctctg (SEQ ID NO: 16)
9
caggtgtccatt (SEQ ID NO: 17)
aatggacacctg (SEQ ID NO: 18)
11
cctacgatatac (SEQ ID NO: 21)
gtatatcgtagg (SEQ ID NO: 22)
12
cgaatgtagagt (SEQ ID NO: 23)
actctacattcg (SEQ ID NO: 24)
13
cggtttgagata (SEQ ID NO: 25)
tatctcaaaccg (SEQ ID NO: 26)
16
gacataaagcga (SEQ ID NO: 31)
tcgctttatgtc (SEQ ID NO: 32)
17
gccatagtctct (SEQ ID NO: 33)
agagactatggc (SEQ ID NO: 34)
18
gctaattcacca (SEQ ID NO: 35)
tggtgaattagc (SEQ ID NO: 36)
20
gttgattctgtc (SEQ ID NO: 39)
gacagaatcaac (SEQ ID NO: 40)
23
ttccacttaggg (SEQ ID NO: 45)
ccctaagtggaa (SEQ ID NO: 46)
25
tttcccttgcta (SEQ ID NO: 49)
tagcaagggaaa (SEQ ID NO: 50)
and wherein each oligonucleotide of the pair is in a separate vial.
3. The kit of claim 2 , wherein the linking agent on one oligonucleotide comprises a biotin, streptavidin, avidin, amino group, thiol group, aldehyde group, hydrazide group, azide group, alkyne group, maleimide group or iodoacetamide group.
4. The kit of claim 2 , wherein one oligonucleotide in each pair of oligonucleotides is coupled to an antibody.
5. The kit of claim 2 , wherein each of the oligonucleotides is modified at the 3′ end.
6. The pair of complementary oligonucleotides of claim 1 , wherein each of the oligonucleotides is modified at the 3′ end.
7. The pair of complementary oligonucleotides of claim 1 , wherein each of the pair of oligonucleotides has a length of 10 to 25 bases.
8. The kit of claim 2 , wherein each of the pair of oligonucleotides has a length of 10 to 25 bases.
9. The pair of complementary oligonucleotides of claim 1 , wherein the linking agent on one oligonucleotide comprises a biotin, streptavidin, avidin, amino group, thiol group, aldehyde group, hydrazide group, azide group, alkyne group, maleimide group or iodoacetamide group.
10. The pair of complementary oligonucleotides of claim 1 , wherein one oligonucleotide in the pair of oligonucleotides is coupled to an antibody.
11. The kit of claim 3 , wherein one oligonucleotide of the complementary oligonucleotide pair is modified with a linking agent comprising biotin and wherein the other oligonucleotide of the complementary oligonucleotide pair is modified with a linking agent comprising streptavidin or avidin.
12. The pair of complementary oligonucleotides of claim 10 , wherein one oligonucleotide of the complementary oligonucleotide pair is modified with a linking agent comprising biotin and wherein the other oligonucleotide of the complementary oligonucleotide pair is modified with a linking agent comprising streptavidin or avidin.
13. The kit of claim 2 , wherein the kit comprises at least three pairs of complementary oligonucleotides.
14. The kit of claim 2 , wherein the kit comprises at least five pairs of complementary oligonucleotides.
15. The kit of claim 2 , wherein the kit comprises at least eight pairs of complementary oligonucleotides.