Solubilized phospholipids for stabilizing nucleic acid polymerases

• US 8,883,474 B2
• Filed: 12/20/2013
• Issued: 11/11/2014
• 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.
• Primary Examiner(s): NOAKES, SUZANNE MARIE
• Application Number:
  US14/136,152
• Publication Number:
  US 20140113349A1
• Time in Patent Office: 326 Days
• Field of Search: None
• 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. A method comprising:

providing a thermostable polymerase in a buffer and a solubilized long-chain phospholipid, wherein the solubilized phospholipid has at least 8 to 30 carbons; and

stabilizing the thermostable polymerase.

2. A method according to claim 1, wherein the solubilized long-chain phospholipid is solubilized by adding an amphipathic molecule.

3. A method according to claim 2, wherein the solubulizer is a lipid having a side chain of no more than 8 carbons.

4. A method according to claim 1, further comprising:

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