Binding reagents that contain small epitope binding molecules
The invention provides binding reagents that contain a plurality of linked small epitope binding molecules that each recognizes a small epitope, such as small epitope antibodies. The combination of small epitope binding molecules in a binding reagent specifically recognizes and binds to a molecule of interest. The binding reagents may be used for such purposes as detection, quantification, identification, and purification of molecules of interest.
1 . A binding reagent comprising a plurality of small epitope binding molecules, wherein said small epitope binding molecules are linked, and wherein each of said small epitope binding molecules recognizes a different small epitope.
2 . A binding reagent according to claim 1 , wherein said plurality of small epitope binding molecules consists of 2, 3, or 4 small epitope binding molecules.
3 . A binding reagent according to claim 1 , wherein said small epitope binding molecules are small epitope antibodies or epitope binding fragments thereof.
4 . A binding reagent according to claim 3 , wherein each small epitope antibody or epitope binding fragment thereof recognizes an epitope consisting of 3, 4, or 5 contiguous amino acids.
5 . A binding reagent according to claim 3 , wherein said plurality of small epitope binding molecules consists of 2, 3, or 4 small epitope antibodies or epitope binding fragments thereof.
6 . A binding reagent according to claim 1 , wherein said small epitope binding molecules are selected from the group consisting of aptamers, molecular imprints, lectins, and capture compounds.
7 . A binding reagent according to claim 1 , wherein the binding reagent binds specifically to a protein.
8 . A binding reagent according to claim 1 , wherein the small epitope binding molecules are linked covalently.
9 . A binding reagent according to claim 8 , wherein the covalent linkage comprises a chemical cross-linking reagent.
10 . A binding reagent according to claim 1 , wherein the small epitope binding molecules are linked by interaction between members of a binding pair.
11 . A binding reagent according to claim 3 , wherein the reagent is recombinantly expressed as a polypeptide with a peptide linker between the antibodies or epitope binding fragments.
12 . A binding reagent according to claim 1 , comprising a detectable label.
13 . A library of binding reagents, comprising a plurality of binding reagents according to claim 1 , wherein each of said binding reagents recognizes a different combination of small epitopes.
14 . A method of characterizing a molecule of interest, said method comprising contacting a sample suspected of comprising the molecule of interest with a binding reagent according to claim 1 , wherein said binding reagent is capable of specifically binding to the molecule of interest, and detecting binding of the molecule of interest to the binding reagent.
15 . A method according to claim 14 , wherein characterizing comprises determining the amount of the molecule of interest in the sample.
16 . A method according to claim 14 , wherein characterizing comprises determining presence or absence of the molecule of interest in the sample.
17 . A method according to claim 14 , wherein the molecule of interest is a biomarker.
18 . An array comprising a plurality of binding reagents according to claim 1 immobilized on a support, wherein each of said binding reagents recognizes a different combination of small epitopes.
19 . A method of making a binding reagent according to claim 1 , comprising linking said small epitope binding molecules with a chemical cross-linking reagent.
20 . A method of making a binding reagent according to claim 1 , comprising linking said small epitope binding molecules via interaction between members of a binding pair.
21 . A method of making a binding reagent according to claim 3 , comprising recombinant expression of a polypeptide comprising said small epitope antibodies or epitope binding fragments thereof.
22 . A kit comprising a binding reagent according to claim 1 .
23 . A composition comprising a binding reagent according to claim 1 .
24 . A pharmaceutical composition comprising a binding reagent according to claim 1 and a pharmaceutically acceptable carrier.
25 . An affinity matrix comprising a binding reagent according to claim 1 attached to a substrate.
26 . A method for purifying a molecule of interest, comprising contacting an affinity matrix according to claim 25 with a sample comprising the molecule of interest, wherein said binding reagent binds to said molecule of interest.
27 . A method according to claim 26 , wherein said molecule of interest is selected from the group consisting of a polypeptide, a polynucleotide, a polysaccharide, and a glycoprotein.