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The use of isothermal titration calorimetry and molecular dynamics to show variability in DNA transfection performance

Paper ID Volume ID Publish Year Pages File Format Full-Text
822 67 2013 9 PDF Available
Title
The use of isothermal titration calorimetry and molecular dynamics to show variability in DNA transfection performance
Abstract

The mechanism causing variability in DNA transfection efficacy for low-molecular-weight pDMAEMA (poly(2-(dimethylamino)ethyl methacrylate) and pDMAEMA-b-pHEMA (poly(2-(dimethyl amino)ethylmethacrylate)-block-poly(2-hydroxyl methacrylate)) has so far remained unclear, apart from the evidence of beneficial effects of the pHEMA grafting. This study has explicitly characterized the electrostatically driven self-assembly process of linear polymethacrylate polymers with DNA-generating nanocarriers for efficient gene transfection. Isothermal titration calorimetry (ITC) showed clear differences in binding-heat profiles of homo-polycationic and pHEMA grafted polymers with DNA. Polyethylene imine, a branched polycationic polymer of 25 kDa with high transfection potential that has previously been successfully used in transfection experiments, demonstrated a heat flow profile that was partly identical to pDMAEMA-b-pHEMA. Computational molecular dynamics (MD) simulated the folding process of polymer in water from a linear to a coiled state: homo-pDMAEMA and pHEMA grafts reduced their overall positive charge accessibility upon folding, down to 45% and 63%, respectively. The homo-pDMAEMA formed the globular conformation more preferably than pHEMA grafts, thus impeding electrostatic interaction with DNA. These findings substantiate the known disadvantage of low-molecular-weight linear polymers compared to higher-molecular-weight polymers in transfection performance; here we have disclosed the ability of a non-cationic chain elongation to be beneficial for the self-assembly process. The combination of MD and ITC has proved to be a suitable approach for carrier–payload interaction studies and may be used to predict the efficacy of a polymer as a nanocarrier from the flexibility of its structure.

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Keywords
Isothermal titration; Calorimetry; Molecular dynamics simulation; Diblock copolymers; Polyelectrolyte chains
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The use of isothermal titration calorimetry and molecular dynamics to show variability in DNA transfection performance
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Publisher
Database: Elsevier - ScienceDirect
Journal: Acta Biomaterialia - Volume 9, Issue 2, February 2013, Pages 4994–5002
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us