Nuclease inhibitor cocktail
View Patent ↗Methods and compositions for inhibiting and/or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease. The invention also relates to kits for the performance of various microbiological procedures, which kits comprise the nuclease inhibitors described herein.
1. A method comprising:
a) obtaining at least a first soluble anti-nuclease antibody;
b) obtaining at least a second soluble anti-nuclease antibody wherein said first and said at least second soluble anti-nuclease antibodies bind to different nucleases;
c) obtaining a composition; and
d) admixing the anti-nuclease antibodies and the composition to form an admixture;
wherein nucleases that may be present in the admixture are inhibited and
wherein at least one of the first and second soluble anti-nuclease antibodies is a monoclonal antibody.
2. The method of claim 1 , wherein the first and the second soluble anti-nuclease antibodies are monoclonal antibodies.
3. The method of claim 1 , wherein the first soluble anti-nuclease antibody is an anti-ribonuclease monoclonal antibody.
4. The method of claim 1 , wherein the first soluble anti-nuclease antibody is an anti-RNase A monoclonal antibody and the second soluble anti-nuclease antibody is an anti-RNase 1 monoclonal antibody.
5. The method of claim 1 , wherein the first soluble anti-nuclease antibody is an anti-RNase 1 monoclonal antibody and the second soluble anti-nuclease antibody is an anti-RNase T1 monoclonal antibody.
6. The method of claim 1 , wherein the first soluble anti-nuclease antibody is an anti-RNase T1 monoclonal antibody and the second soluble anti-nuclease antibody is an anti-RNase A monoclonal antibody.
7. The method of claim 1 , wherein the first soluble anti-nuclease antibody is an anti-deoxyribonuclease monoclonal antibody.
8. The method of claim 1 , comprising obtaining at least an anti-RNase A antibody, an anti-RNase 1 antibody, and an anti-RNase T1 antibody and admixing them with the composition, wherein at least one of the anti-RNase A antibody, the anti-RNase 1 antibody, and the anti-RNase T1 antibody is a monoclonal antibody.
9. The method of claim 1 , comprising obtaining at least an anti-RNase A monoclonal antibody, an anti-RNase 1 monoclonal antibody, and an anti-RNase T1 monoclonal antibody and admixing them with the composition.
10. A method comprising:
a) obtaining at least a first nuclease inhibitor further defined as a soluble anti-nuclease monoclonal antibody;
b) obtaining at least a second nuclease inhibitor further defined as a non-antibody nuclease inhibitor;
c) obtaining a composition; and
d) admixing the first nuclease inhibitor, the second nuclease inhibitor, and the composition to fonn an admixture; wherein nucleases that may be present in the admixture are inhibited.
11. The method of claim 10 , wherein the soluble anti-nuclease monoclonal antibody is an anti-ribonuclease monoclonal antibody.
12. The method of claim 11 , wherein the soluble anti-ribonuclease monoclonal antibody is an anti-RNase 1 monoclonal antibody.
13. The method of claim 11 , wherein the soluble anti-ribonuclease monoclonal antibody is an anti-RNase T1 monoclonal antibody.
14. The method of claim 11 , wherein the soluble anti-ribonuclease monoclonal antibody is an anti-RNase A monoclonal antibody.
15. The method of claim 10 , wherein the second nuclease inhibitor is human placental ribonuclease inhibitor.
16. A kit comprising:
a. a first soluble anti-ribonuclease monoclonal antibody that binds to at least a first ribonuclease;
b. at least one of:
a second soluble anti-nuclease antibody that binds to a nuclease that is different from the first nuclease; and
a non-antibody ribonuclease inhibitor; and
c. a reagent used in molecular biology.
17. The kit of claim 16 comprising:
a. a first soluble anti-ribonuclease monoclonal antibody that binds to at least a first ribonuclease;
b. a second soluble anti-nuclease antibody that binds to a nuclease that is different from the first nuclease; and
c. a reagent used in molecular biology.
18. The kit of claim 16 comprising:
a. a first soluble anti-ribonuclease monoclonal antibody that binds to at least a first ribonuclease;
b. a non-antibody ribonuclease inhibitor; and
c. a. reagent used in molecular biology.
19. The kit of claim 16 wherein the first soluble anti-ribonuclease monoclonal antibody is an anti-RNase A monoclonal antibody, an anti-RNase T1 nionoclonal antibody, or an anti-RNase 1 monoclonal antibody.
20. The kit of claim 18 wherein the non-antibody ribonuclease inhibitor is placental ribonuclease inhibitor.