Polyanionic acids to improve recovery and minimize system loss

Patent metadata

Field	Value
Publication number	US-12359191-B2
Application number	US-202418414679-A
Country	US
Kind code	B2
Filing date	Jan 17, 2024
Priority date	Dec 5, 2019
Publication date	Jul 15, 2025
Grant date	Jul 15, 2025

Abstract

The present technology relates to a method of separating a sample comprising oligonucleotides. The method includes injecting a polyphosphonic acid at a concentration of between about 0.01 M to about 1 M into the sample comprising oligonucleotides. The method also includes flowing the sample and polyphosphonic acid through a liquid chromatography column and separating the oligonucleotides.

First claim

What is claimed is:

1. A method of separating a sample comprising oligonucleotides comprising:
injecting a polyphosphonic acid at a concentration of between about 0.01 M to about 1 M into the sample comprising oligonucleotides;
flowing the sample and polyphosphonic acid through a liquid chromatography column; and
separating the oligonucleotides.

2. The method of claim 1 , wherein the concentration of polyphosphonic acid is between about 0.01 M to about 0.1 M.

3. The method of claim 2 , wherein the concentration of polyphosphonic acid is between about 20 mM to about 50 mM.

4. The method of claim 3 , wherein the concentration of polyphosphonic acid is between about 30 mM to about 40 mM.

5. The method of claim 1 , wherein between about 1 pg to about 1 mg of the polyphosphonic acid is injected into the sample.

6. The method of claim 1 , wherein the polyphosphonic acid is etidronic acid.

7. The method of claim 1 , wherein the polyphosphonic acid is nitrilotri (methylphosphonic acid).

8. The method of claim 1 , wherein walls of the liquid chromatography column are formed of metal.

9. The method of claim 1 , wherein the sample comprises phosphorylated oligonucleotides.

10. The method of claim 9 , wherein the phosphorylated oligonucleotides comprise nucleotides and at least one nucleotide is adenosine triphosphate.

11. The method of claim 1 , wherein the polyphosphonic acid is etidronic acid at pH of 8.5 and the oligonucleotide comprises nucleotides and at least one nucleotide is adenosine triphosphate.

12. The method of claim 1 , wherein the polyphosphonic acid is a bisphosphonic acid.

13. The method of claim 12 , wherein the bisphosphonic acid is selected from the group consisting of clodronic acid, pamidronic acid, alendronic acid, neridronic acid, and olpadronic acid.

Assignees

• Waters Technologies Corp

Inventors

• Donegan Michael
• Gilar Martin
• Lauber Matthew
• Mccall Scott
• Iraneta Pamela
• Smith Kerri

Classifications

• C12N2310/315

  Phosphorothioates · CPC title

• C12N2310/11

  Antisense · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• G01N2030/8836

  involving saccharides · CPC title

• G01N2030/8831

  involving peptides or proteins · CPC title