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Team:Alberta/References/Publications/Shear-induced degradation of plasmid DNA
From 2009.igem.org
C. S. Lengsfeld and T. J. Anchordoquy
J Pharm Sci. 2002 Jul;91(7):1581-9
Abstract: The majority of gene therapy clinical trials use plasmid DNA that is
susceptible to shear-induced degradation. Many processing steps in the extraction, purification,
and preparation of plasmid-based therapeutics can impart significant shear
stress that can fracture the phosphodiester backbone of polynucleotides, and reduce
biological activity. Much of the mechanistic work on shear degradation of DNA was
conducted over 30 years ago, and we rely heavily on this early work in an attempt to
explain the empirical observations of more recent investigations concerning the
aerosolization of plasmids. Unfortunately, the sporadic reports of shear degradation
in the literature use different experimental systems, making it difficult to quantitatively
compare results and reach definitive mechanistic conclusions. In this review, we
describe the forces imparted to DNA during shear stress, and use published data to
quantitatively evaluate their relative effects. In addition, we discuss the effects of
molecular weight, strain rate, particle size, flexibility, ionic strength, gas–liquid
interfaces, and turbulence on the fluid flow degradation of supercoiled plasmid DNA.
Finally, we speculate on computational methods that might allow degradation rates in
different experimental systems to be predicted.
Link: [http://www.ncbi.nlm.nih.gov/pubmed/12115820 NCBI]
