Patent ID: 8541559

Claim:
A process for producing an nucleotide composition comprising aggregated oligonucleotides, said process comprising the steps of: (a) providing at least two oligonucleotides in solution I, wherein said oligonucleotides comprise at their 5′ end at least 3 and at most 15 guanosine entities and at their 3′ end at least 3 and at most 15 guanosine entities, wherein each oligonucleotide of said oligonucleotides comprise 20 to 40 nucleotides, and wherein said solution I comprises a pH of 10 to 13; (b) disaggregating said oligonucleotides, wherein said disaggregating comprises the steps of (i) adjusting the temperature of solution I to temperature I, wherein said temperature I is 20 to 70° C.; (ii) incubating said oligonucleotides in said solution I at said temperature I, wherein said incubating is performed until said oligonucleotides comprise a relative peak start time above 110%, and wherein said relative peak start time is determined in size exclusion HPLC with the capsid of an RNA bacteriophage as the standard; and (iii) adjusting the temperature of said solution I to temperature II, wherein said temperature II is 0 to 10° C.; (c) adjusting the pH of said solution I to 5 to 8; and (d) aggregating said oligonucleotides, wherein said aggregating comprises the steps of: (i) providing said oligonucleotides in said solution I and generating a solution II, wherein said solution II further comprises at least 20 mM of a cation, wherein said cation is selected from the group consisting of Na + , NH 4 + , Li + , Ca 2+ , and Mg 2+ ; (ii) adjusting the temperature of solution II to temperature III, wherein said temperature III is 50 to 99° C.; (iii) incubating said oligonucleotides in solution II at temperature III, wherein said incubating is performed until said oligonucleotides comprise a relative peak start time of 80 to 110%, wherein said relative peak start time is determined in size exclusion HPLC with the capsid of an RNA bacteriophage as the standard; and (iv) adjusting the temperature of solution II to temperature IV, wherein said temperature IV is 0 to 10° C.