Solubilized Phospholipids for Stabilizing Nucleic Acid Polymerases

• US 20140113349A1
• Filed: 12/20/2013
• Published: 04/24/2014
• Est. Priority Date: 05/02/2011
• Status: Active Grant

• Current Assignee: New England Biolabs Incorporated
• Original Assignee: New England Biolabs Incorporated
• Inventors: Johnson, Donald, Evans, Thomas C. Jr.
• Granted Patent: US 8,883,474 B2
• Time in Patent Office: Days
• US Class Current: 435/188
• CPC Class Codes: C12N 9/1241 Nucleotidyltransferases (2.7.7)
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2521/101 DNA polymerase
• CPC Class Codes: C12Q 2527/127 the enzyme inhibitor or activator used

Abstract

Compositions and methods are provided that relate to solubilized phospholipids and their use in stabilizing nucleic acid polymerases. For example, a phospholipid with a tail containing at least 8 carbons can be solubilized in the presence of an amphipathic molecule.

Claims

1-7. -7. (canceled)

8. A method comprising:

providing a nucleic acid polymerase in a buffer and a solubilized long-chain phospholipid, the solubilized phospholipid having at least 8 carbons; and

stabilizing the nucleic acid polymerase.

9. A method according to claim 8, wherein the solubilized long-chain phospholipid is solubilized by adding an amphipathic molecule.

10. A method according to claim 9, wherein the solubulizer is a lipid having a side chain of no more than 8 carbons.

11. A method according to claim 8, further comprising:

enhancing the activity of the polymerase in the presence of the solublilized phospholipid as can be measured in a replication assay by incorporation of deoxynucleoside triphosphate.

12-14. -14. (canceled)