Purification and administration of DNA repair enzymes

Details Purification and administration of DNA repair enzymes Purification and administration of DNA repair enzymes

1993-02-05

1993-12-21

Patterson, Jr., Charles L.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

435196, 424 945, 424 946, 424450, C12N 910, C12N 916, A61K 3752, A61K 3754

Patent

active

052720799

ABSTRACT:
Methods for purifying DNA repair enzymes are provided in which an aqueous solution of a DNA repair enzyme in an impure state is applied to a molecular sieve column having an exclusion limit which will retard the DNA repair enzyme but will not retard contaminants larger than the DNA repair enzyme. The DNA repair enzyme in an enhanced state of purity is eluted isocratically from the molecular sieve column in an elution buffer and applied directly to a DNA affinity column in the same buffer without intermediate dialysis, ultrafiltration, or other procedures. The DNA repair enzyme is eluted from the DNA affinity column using, for example, a salt gradient. The method is rapid, inexpensive, simple to perform, and has been found to produce a homogeneous final product. In accordance with other aspects of the invention, the purified DNA repair enzymes are encapsulated in liposomes and administered to living cells in situ. This form of administration has been found to be non-toxic to the cells and to result in increased incision of damaged DNA, enhanced DNA repair synthesis, and increased cell survival after exposure to ultraviolet light. In certain preferred embodiments, DNA repair enzymes are administered in pH sensitive liposomes.

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