FUNCTIONAL LIGANDS TO TARGET MOLECULES AND METHODS
The present invention relates functional ligands to target molecules, particularly to functional nucleic acids and modifications thereof, and to methods for simultaneously generating, for example, numerous different functional biomolecules, particularly to methods for generating numerous different functional nucleic acids against multiple target molecules simultaneously. The present invention further relates to functional ligands which bind with affinity to target molecules.
1 . An artificial ligand binding to CD63 comprising a non-naturally occurring nucleic acid sequence having substantial homology or identity to a sequence selected from the group consisting of SEQ IDs 122-183.
2 . The artificial ligand of claim 1 further comprising at least one modified nucleotide.
3 . An artificial ligand binding to prealbumin comprising a non-naturally occurring nucleic acid sequence having substantial homology or identity to a sequence selected from the group consisting of SEQ IDs 1-62.
4 . The artificial ligand of claim 3 further comprising at least one modified nucleotide.
5 . A method for selecting nucleic acid aptamers from a pool of candidates comprising:
providing a plurality of non-nucleic acid target molecules with each specie of target molecule being substantially immobilized in a target spot at a known spatial address on a substrate;
contacting under binding conditions a pool with said substrate, said pool comprising a plurality of randomized and artificially synthesized single-stranded nucleic acids, each of said single-stranded nucleic acids being non-naturally occurring and comprising a target binding region which is unique in said plurality and a hybridizing region which is constant in said pool;
washing said substrate to remove any of said single-stranded nucleic acids which are not bound to at least one of said target molecules;
applying an identifier to each of said target spots under hybridization conditions, each of said identifiers comprising a nucleic acid primer capable of hybridizing to said hybridizing region and having a sequence which indicates which said spatial address said identifier was applied;
performing a nucleic acid extension on each said nucleic acid primer to form an extension product comprising said nucleic acid primer and the reverse-complement of said target binding region of the single-stranded nucleic acid to which said nucleic acid primer is hybridized; and
collecting and sequencing said extension products to correlate which of said single-stranded nucleic acids was bound at a particular spatial address via said sequence of said nucleic acid primer in said extension product.
6 . The method of claim 5 , wherein said target spots are present as a high content protein array.