Affinity tag nucleic acid and protein compositions, and processes for using same
The present invention concerns compositions and processes that use affinity tags for isolating, and detecting or quantifying analytes, including nucleic acids, proteins and polypeptides. Compositions include nucleic acid compositions and protein compositions with affinity binding pairs, including metal binding peptides and immobilized metals, or peptide affinity groups.
1. A composition of matter, comprising:
a chimeric molecule comprising
a polynucleic acid portion comprising one or more energy transfer donors or energy transfer acceptors, and
a peptide affinity tag portion covalently attached to the polynucleic acid portion;
a first polynucleic acid molecule hybridized to the polynucleic acid portion of the chimeric molecule; and
a second polynucleic acid molecule hybridized to the first polynucleic molecule and not to the polynucleic acid portion of the chimeric molecule, said second polynucleic acid comprising one or more energy transfer donors or energy transfer acceptors,
wherein when the polynucleic acid portion comprises one or more energy transfer donors, the second polynucleic acid molecule comprises one or more energy transfer acceptors and when the polynucleic acid portion comprises one or more energy transfer acceptors, the second polynucleic acid molecule comprises one or more energy transfer donors.
2. The composition of matter of claim 1 , wherein the polynucleic acid portion of the chimeric molecule and the peptide affinity tag portion of the chimeric molecule are covalently attached through a linker arm.
3. The composition of matter of claim 1 , wherein the peptide affinity tag portion comprises a metal-binding peptide affinity tag.
4. The composition of matter of claim 3 , wherein the polynucleic acid portion of the chimeric molecule and the peptide affinity tag portion of the chimeric molecule are covalently attached through a linker arm.
5. The composition of claim 3 , further comprising an immobilized metal wherein the metal-binding affinity tag is bound to the immobilized metal.
6. The composition of matter of claim 5 , wherein the polynucleic acid portion of the chimeric molecule and the peptide affinity tag portion of the chimeric molecule are covalently attached through a linker arm.