Large-volume methacrylate monolith for plasmid purification : process engineering approach to synthesis and application

Danquah, Michael K. & Forde, Gareth M. (2008) Large-volume methacrylate monolith for plasmid purification : process engineering approach to synthesis and application. Journal of Chromatography A, 1188(2), pp. 227-233.

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Abstract

The extent of exothermicity associated with the construction of large-volume methacrylate monolithic columns has somewhat obstructed the realisation of large-scale rapid biomolecule purification especially for plasmid-based products which have proven to herald future trends in biotechnology. A novel synthesis technique via a heat expulsion mechanism was employed to prepare a 40 mL methacrylate monolith with a homogeneous radial pore structure along its thickness. Radial temperature gradient was recorded to be only 1.8 °C. Maximum radial temperature recorded at the centre of the monolith was 62.3 °C, which was only 2.3 °C higher than the actual polymerisation temperature. Pore characterisation of the monolithic polymer showed unimodal pore size distributions at different radial positions with an identical modal pore size of 400 nm. Chromatographic characterisation of the polymer after functionalisation with amino groups displayed a persistent dynamic binding capacity of 15.5 mg of plasmid DNA/mL. The maximum pressure drop recorded was only 0.12 MPa at a flow rate of 10 mL/min. The polymer demonstrated rapid separation ability by fractionating Escherichia coli DH5α-pUC19 clarified lysate in only 3 min after loading. The plasmid sample collected after the fast purification process was tested to be a homogeneous supercoiled plasmid with DNA electrophoresis and restriction analysis.

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24 citations in Web of Science®

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ID Code: 81555
Item Type: Journal Article
Refereed: Yes
Keywords: Anion-exchange, Large-volume, Methacrylate monolith, Plasmid DNA, Temperature, Biomolecules, Biotechnology, DNA, Exothermic reactions, Ion exchange, Pore structure, Purification, Thermal gradients, Anion exchange, Large volume methacrylate monolith, Acrylics, methacrylic acid, article, binding affinity, chemical engineering, chromatography, DNA purification, DNA supercoiling, DNA synthesis, electrophoresis, flow rate, high temperature, low temperature, polymerization, porosity, priority journal, separation technique, temperature dependence, thickness, Electrophoresis, Agar Gel, Escherichia coli, Methacrylates, Microscopy, Electron, Scanning, Plasmids, Polymers, Pressure, Time Factors
DOI: 10.1016/j.chroma.2008.02.045
ISSN: 0021-9673
Divisions: Current > Schools > School of Chemistry, Physics & Mechanical Engineering
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 2008 Elsevier B.V.
Deposited On: 06 Feb 2015 00:29
Last Modified: 11 Feb 2015 04:33

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