초록 ▼

The present invention discloses improved methods for oligonucleotide synthesis and purification. In the methods of the invention, a half-life is determined for deprotection of the 5′-OH protecting group at the 5′-terminus of an oligonucleotide post-synthesis. The half-life is used to d

The present invention discloses improved methods for oligonucleotide synthesis and purification. In the methods of the invention, a half-life is determined for deprotection of the 5′-OH protecting group at the 5′-terminus of an oligonucleotide post-synthesis. The half-life is used to determine an optimal reaction time for removal of the 5′-OH protecting group. The methods of the invention are amenable to the large-scale synthesis and purification of oligonucleotides.

대표청구항 ▼

1. A method for improving deprotection of an acid-labile 5′-OH protecting group-containing oligonucleotide comprising:determining the half-life for said acid-labile 5′-OH protecting group-containing oligonucleotide in an acid solution capable of removing the acid-labile 5′-OH pr

1. A method for improving deprotection of an acid-labile 5′-OH protecting group-containing oligonucleotide comprising:determining the half-life for said acid-labile 5′-OH protecting group-containing oligonucleotide in an acid solution capable of removing the acid-labile 5′-OH protecting group from said oligonucleotide, andreacting said acid-labile 5′-OH protecting group-containing oligonucleotide in said acid solution for approximately 5 to approximately 20 half-lives;wherein said acid solution comprises acetic acid, dichloracetic acid, trichloroacetic acid, zinc bromide or boron trifluoride etherate. 2. The method of claim 1 wherein said oligonucleotide is reacted in said acid solution for approximately 10 to approximately 15 half-lives. 3. The method of claim 2 wherein said half-life is determined from a standard curve of pH values vs. half-lives. 4. The method of claim 2 wherein said half-life is determined by evaluating exponential decay of said protecting group-containing oligonucleotide. 5. The method of claim 2 wherein said deprotection is detritylation and said acid-labile 5′-OH protecting group is a trityl group. 6. The method of claim 4 wherein said trityl group is di-p-methoxytrityl. 7. A method for deprotecting an acid-labile 5′-OH protecting group-containing oligonucleotide comprising:purifying said acid-labile 5′-OH protecting group-containing oligonucleotide,determining a half-life for said acid-labile 5′-OH protecting group-containing oligonucleotide in an acid solution capable of removing the acid-labile 5′-OH protecting group from said oligonucleotide, andreacting said acid-labile 5′-OH protecting group-containing oligonucleotide in said acid solution for approximately 10 to approximately 15 half-lives;wherein said acid solution comprises acetic acid, dichloracetic acid, trichloroacetic acid, zinc bromide or boron trifluoride etherate. 8. The method of claim 7 wherein said purifying is done by high performance liquid chromatography or strong ion exchange chromatography. 9. The method of claim 7 wherein a solvent used in said high performance liquid chromatography contains sodium acetate. 10. The method of claim 7 wherein said half-life is determined from a standard curve of pH values vs. half-lives. 11. The method of claim 7 wherein said half-life is determined by evaluating exponential decay of said protecting group-containing oligonucleotide. 12. The method of claim 7 wherein said deprotection is detritylation and said acid-labile 5′-OH protecting group is a trityl group. 13. The method of claim 12 wherein said trityl group is di-p-methoxytrityl. 14. The method of claim 1 wherein said acid solution comprises acetic acid, dichloracetic acid, or trichloroacetic acid. 15. The method of claim 1 wherein said acid solution comprises dichloracetic acid. 16. The method of claim 7 wherein said acid solution comprises acetic acid, dichloracetic acid, or trichloroacetic acid. 17. The method of claim 7 wherein said acid solution comprises dichloracetic acid.