Methods for rapid purification of nucleic acids for subsequent analysis by mass spectrometry by solution capture

• US 8,476,415 B2
• Filed: 04/16/2012
• Issued: 07/02/2013
• Est. Priority Date: 05/13/2003
• Status: Expired due to Fees

• Current Assignee: Ibis Biosciences Incorporated
• Original Assignee: Ibis Biosciences Incorporated
• Inventors: Hofstadler, Steven A., Cummins, Lendell L.
• Primary Examiner(s): Chunduru, Prabha
• Application Number:
  US13/447,623
• Publication Number:
  US 20120207655A1
• Time in Patent Office: 442 Days
• Field of Search: None
• US Class Current: 536/22.1
• CPC Class Codes: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/143 Magnetism, e.g. magnetic label
• CPC Class Codes: C12Q 2563/155 Particles of a defined size, e.g. nanoparticles
• CPC Class Codes: C12Q 2565/627 being a mass spectrometer
• CPC Class Codes: H01J 49/165 Electrospray ionisation

Abstract

The present invention provides a method for rapid solution capture purification of nucleic acids for subsequent analysis by electrospray mass spectrometry which is efficient and cost-effective relative to existing methods. The present invention also provides kits useful for practicing rapid solution capture of nucleic acids so that purified samples are in condition for analysis by electrospray mass spectrometry.

Claims

1. A kit for purification of nucleic acids comprising:

a resin source plate containing anion exchange resin, wherein said anion exchange resin comprises amine-terminated magnetic beads, and wherein each of said amine-terminated magnetic beads comprises a magnetic bead linked to at least one amine molecule;

a sample mixing plate;

a filter plate or a microtube rack comprising a plurality of micro filter tubes;

one or more resin wash solutions; and

an electrospray ionization-mass spectrometry-compatible elution buffer, wherein said elution buffer does not comprise a metal cation salt.

2. The kit of claim 1 wherein said filter plate is a 96-well plate or a 384-well plate.

3. The kit of claim 1 wherein said anion exchange resin comprises a weak anion exchange functional group.

4. The kit of claim 1 wherein said anion exchange resin comprises ProPac™

WAX.

5. The kit of claim 1 wherein said anion exchange resin comprises a weak anion exchange functional group bound to a magnetic bead.

6. The kit of claim 1 wherein said weak anion exchange resin is ProPac™

WAX.

7. The kit of claim 1 wherein said one or more wash solutions comprises about 50 mM to about 200 mM ammonium acetate or about 50 mM to about 200 mM ammonium bicarbonate.

8. The kit of claim 1 wherein said at least one anion exchange wash buffer comprises about 20% to about 50% methanol.

9. The kit of claim 1 wherein said ESI-MS compatible elution buffer comprises about 2% to about 6% ammonium hydroxide.

10. The kit of claim 1 wherein said ESI-MS compatible elution buffer comprises about 10 mM to about 50 mM imidazole, about 10 mM to about 50 mM piperidine and about 20% to about 50% methanol.

11. The kit of claim 1 wherein said ESI-MS compatible elution buffer comprises about 10 mM to about 50 mM imidazole, about 10 mM to about 50 mM piperidine and about 20% to about 50% methanol.