Multi-component inhibitors of nucleic acid polymerases
AssigneeInvitrogen Corporation
InventorsJohn Bishop, Jun Lee
Life of this patent
Estimating the term…
Abstract
The present invention provides multi-component inhibitors of nucleic acid polymerases, methods of making, and methods of using same. One component of the multi-component inhibitor is a molecule that binds to a polymerase (i.e., a polymerase-binding molecule (PBM)), but does not thereby substantially inhibit its polymerase activity. Another component is a molecule or complex of molecules that binds to a PBM (i.e., a PBM-binding molecule). The combination of the PBM and PBM-binding molecule/complex substantially inhibits polymerase activity.
Claims28
A composition comprising:
- (a) a nucleic acid polymerase;
- (b) a polymerase-binding molecule (PBM) that binds to said polymerase and does not substantially inhibit the polymerase activity of said polymerase;
- and
- (c) a PBM-binding molecule/complex that binds to said PBM, wherein binding of the PBM and PBM-binding molecule or complex together substantially inhibits the polymerase activity of said polymerase.
6-9. (canceled)
12-17. (canceled)
(canceled)
A composition comprising:
- (a) a thermostable nucleic acid polymerase;
- (b) a polymerase-binding antibody (PBA) that binds to said polymerase and does not substantially inhibit the polymerase activity of said polymerase;
- and
- (c) a PBA-binding molecule/complex that binds to said PBA, wherein binding of the PBA and PBA-binding molecule/complex together substantially inhibits the polymerase activity of said polymerase at a temperature less than about 40° C., and wherein binding of the PBA and PBA-binding molecule/complex together does not substantially inhibit the polymerase activity of said polymerase at a temperature greater than about 40° C.
22-23. (canceled)
A method of inhibiting the polymerase activity of a nucleic acid polymerase comprising contacting said polymerase with a PBM and a PBM-binding molecule/complex, wherein the binding of said PBM does not substantially inhibit polymerase activity of said polymerase, and wherein the binding of said PBM and said PBM-binding molecule/complex together substantially inhibit the polymerase activity of said polymerase.
30-31. (canceled)
A method for synthesizing a nucleic acid molecule, comprising:
- (a) contacting a template nucleic acid with a composition comprising a thermostable nucleic acid polymerase, one or more nucleoside and/or deoxynucleoside triphosphates, a polymerase binding molecule (PBM) that does not substantially inhibit the polymerase activity of the polymerase, and a PBM-binding molecule/complex under conditions where the PBM and the PBM-binding molecule/complex together substantially inhibit the polymerase activity of the polymerase;
- and
- (b) bringing the resulting mixture to a temperature sufficient relieve polymerase inhibition.
37-38. (canceled)
Dependencies are inferred from the claim text.