Method of preparing an adduct

Abstract

A method for identifying one or several molecular structure(s) having a high-affinity for a target of interest, the molecular structure(s) each including one nucleotide chain onto which is hybridized at least one PNA-encoded molecule.

1 - 17 . (canceled) 
     
     
         18 . A method for identifying a molecular structure having high-affinity for a target of interest in solution, said molecular structure(s) each comprising one nucleotide chain onto which is hybridized at least two PNA-encoded molecules, the method comprising:
 (a) providing at least two libraries of peptide nucleic acid (PNA)-encoded molecules, a first library having PNA-encoded molecules appended at the N-terminus and a second library having PNA-encoded molecules appended at the C-terminus, and a library of nucleotide chains;   (b) hybridizing said at least two libraries of PNA-encoded molecules with said library of nucleotide chains, resulting in a library of PNA-encoded molecule/nucleotide chain hybrids;   (c) bringing into contact the resulting library of PNA-encoded molecule/nucleotide chain hybrids with said target of interest;   (d) selecting a set of the fittest PNA-encoded molecule/nucleotide chain hybrids for the target of interest, based on highest affinity;   (e) amplifying the nucleotide chains obtained from said fittest PNA molecule/nucleotide chain hybrids;   (f) hybridizing said amplified nucleotide chains with at least two libraries of PNA-encoded molecules and repeating steps (c) to (e), step (f) being repeated until a convergence towards one or more consensus nucleotide chain sequences is obtained; and   (g) identifying said nucleotide chain(s) obtained in step (f), each of said nucleotide chain(s) corresponding to at least one PNA-encoded molecule of the at least two libraries of PNA-encoded molecules recited in step (a).   
     
     
         19 . The method according to  claim 18 , wherein step (f) is repeated less than 20 times. 
     
     
         20 . The method according to  claim 18 , wherein the nucleotide chain comprises DNA. 
     
     
         21 . The method according to  claim 18 , further comprising
 (h) determining a consensus structure provided by the molecules of the PNA-encoded molecules corresponding to the nucleotide chain(s) identified in step (g), said consensus structure having a high-affinity to the target of interest.   
     
     
         22 . The method according to  claim 18 , wherein the PNA-encoded molecules appended at the N-terminus are obtained in a split and mix fashion. 
     
     
         23 . The method according to  claim 18 , wherein amplifying the nucleotide chains in step (e) comprises conducting PCR utilizing: a 5′ primer containing a tag, a functionalized solid support having affinity for said tag, and a 3′ primer optionally containing a fluorophore. 
     
     
         24 . The method according to  claim 18 , wherein the PNA-encoded molecules appended at the N-terminus and the PNA-encoded molecules appended at the C-terminus hybridize contiguously to the nucleotide chain. 
     
     
         25 . The method according to  claim 18 , wherein the PNA-encoded molecules comprise a nucleic acid of 10 to 20 nucleotides. 
     
     
         26 . The method according to  claim 18 , wherein the PNA-encoded molecules comprise a nucleic acid sequence of 14 nucleotides. 
     
     
         27 . The method according to  claim 26 , wherein the nucleotide chains in the library of nucleotide chains comprise a nucleic acid sequence of at least 28 nucleotides that is complementary to a PNA-encoded molecule appended at the N-terminus and a PNA-encoded molecule appended at the C-terminus, and the nucleotide chains further comprise nucleic acid sequences flanking said at least 28 nucleotides, the flanking sequences being complementary to 5′ and 3′ PCR primers. 
     
     
         28 . The method according to  claim 18 , wherein the at least two libraries of PNA-encoded molecules in step (f) have a content identical to that of said at least two libraries of PNA-encoded molecules provided in step (a). 
     
     
         29 . The method according to  claim 18 , wherein the molecule of the PNA-encoded molecule is selected from the group consisting of amino acid, peptides, peptoids, antibodies, and heterocyclic compounds. 
     
     
         30 . The method according to  claim 18 , wherein the target of interest is selected from the group consisting of proteins, protein complexes, receptors, enzymes, kinases, proteases, antibodies and antigens. 
     
     
         31 . The method according to  claim 18 , wherein the PNA-encoded molecule comprises a saccharide. 
     
     
         32 . The method according to  claim 18 , wherein step (f) is repeated less than 5 times.