Protocol for optimizing purification of enzymatically active human telomerase
OwnerScott Weinrich, Edward Atkinson +3inventors
InventorsScott Weinrich, Edward Atkinson, Serge Lichtsteiner, Alain Vasserot, Ronald Pruzan
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Abstract
This invention provides purified telomerase and methods of purifying it. The methods involve the use of several sequential steps, including the use of matrices that bind molecules bearing negative charges, matrices that bind molecules bearing positive charges, intermediate-selectivity matrices, methods that separate molecules based on their size, shape, or buoyant density, and by affinity purification.
Claims20
A protocol for optimizing purification of human telomerase, comprising:
- a) obtaining a solution of human telomerase;
- b) measuring telomerase enzyme activity in the human telomerase solution;
- c) combining the solution with a test affinity agent;
- d) recovering mammalian telomerase that has bound to the test affinity agent;
- e) measuring telomerase enzyme activity recovered from the affinity agent;
- f) determining enrichment of the telomerase enzyme activity (in terms of telomerase activity per weight of protein) that results from binding and recovery of the human telomerase from the affinity agent;
- and
- g) selecting the affinity agent tested in c) for use in purifying human telomerase in preference to other agents that are less effective for enriching the telomerase enzyme activity by the same criteria.
A protocol for optimizing purification of human telomerase, comprising:
- a) obtaining a solution of human telomerase;
- b) measuring telomerase enzyme activity in the human telomerase solution;
- c) enriching human telomerase in the solution by chromatographic separation using a test separation matrix;
- d) combining the solution with an affinity agent;
- e) recovering mammalian telomerase that has bound to the affinity agent;
- f) measuring telomerase enzyme activity recovered from the affinity agent;
- g) determining enrichment of the telomerase enzyme activity (in terms of telomerase activity per weight of protein) as a result of steps c), d), and e);
- and
- h) identifying the separation matrix used in c) as improving effectiveness of said affinity agent in the purification of human telomerase if said matrix improves enrichment of telomerase enzyme activity recovered from the affinity agent.
A protocol for optimizing purification of human telomerase, comprising:
- a) obtaining a solution of human telomerase;
- b) measuring telomerase enzyme activity in the human telomerase solution;
- c) enriching human telomerase in the solution by a test combination of sequential chromatographic separations using different matrices;
- d) combining the solution with an affinity agent;
- e) recovering mammalian telomerase that has bound to the affinity agent;
- f) measuring telomerase enzyme activity recovered from the affinity agent;
- g) determining enrichment of the telomerase enzyme activity (in terms of telomerase activity per weight of protein) as a result of steps c), d), and e);
- and
- h) identifying the combination of chromatographic separation matrices tested in c) as improving effectiveness of said affinity agent in the purification of human telomerase if said combination improves enrichment of telomerase enzyme activity recovered from the affinity agent.
Dependencies are inferred from the claim text.