Stabilization of polyethyleneimine-deoxyribonucleic acid complex size and activity
AssigneeJuno Therapeutics, Inc.
InventorsBrynn Olden, Robert Barnes, Elizabeth Larimore, Julie Shi
Life of this patent
Estimating the term…
Abstract
Disclosed are methods and systems for producing polymer-DNA nanoparticles of a predetermined size. In one example, a method includes mixing together a first solution comprising deoxyribonucleic acid (DNA) with a second solution comprising a cationic polymer to obtain a polyplex solution, and at a predetermined time subsequent to mixing together the first solution and the second solution, adding a polyplex stabilizing agent to stabilize the size of the polyplex. In this way, transfection efficacy of the polymer-DNA nanoparticles may be improved, in particular with reference to transfection of suspension cells for production of viral vectors.
Claims21
A method of transfecting a population of cells with a polymer-DNA complex, comprising:
- (a) mixing together a first solution comprising deoxyribonucleic acid (DNA) with a second solution comprising a cationic polymer, wherein the cationic polymer is polyethyleneimine (PEI), to obtain a polyplex solution comprising a PEI-DNA complex;
- (b) at a first predetermined time subsequent to mixing together the first solution and the second solution, adding a polyplex stabilizing agent to the polyplex solution to obtain a stabilized PEI-DNA complex;
- and
- (c) at a second predetermined time subsequent to adding the polyplex stabilizing agent to the polyplex solution, transfecting the population of cells with the stabilized PEI-DNA complex;
- wherein the second predetermined time is between about 30 minutes and about 18 hours, and
- wherein the polyplex stabilizing agent is a human serum albumin (HSA).
Dependencies are inferred from the claim text.