A thermal expulsion approach to homogeneous large-volume methacrylate monolith preparation; enabling large-scale rapid purification of biomolecules

Numerous efforts have been dedicated to the synthesis of large-volume methacrylate monoliths for large-scale biomolecules purification but most were obstructed by the enormous release of exotherms during preparation, thereby introducing structural heterogeneity in the monolith pore system. A significant radial temperature gradient develops along the monolith thickness, reaching a terminal temperature that supersedes the maximum temperature required for structurally homogenous monoliths preparation. The enormous heat build-up is perceived to encompass the heat associated with initiator decomposition and the heat released from free radical-monomer and monomer-monomer interactions. The heat resulting from the initiator decomposition was expelled along with some gaseous fumes before commencing polymerization in a gradual addition fashion. Characteristics of 80 mL monolith prepared using this technique was compared with that of a similar monolith synthesized in a bulk polymerization mode. An extra similarity in the radial temperature profiles was observed for the monolith synthesized via the heat expulsion technique. A maximum radial temperature gradient of only 4.3°C was recorded at the center and 2.1°C at the monolith peripheral for the combined heat expulsion and gradual addition technique. The comparable radial temperature distributions obtained birthed identical pore size distributions at different radial points along the monolith thickness.

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

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.

Process engineering approach to large-volume methacrylate monolith synthesis for plasmid purification

The construction of large?volume methacrylate monolithic columns for preparative-scale plasmid purification is obstructed by the enormous release of exotherms, thus introducing structural heterogeneity in the monolith pore system. A remarkable radial temperature gradient develops along the monolith thickness, reaching a terminal temperature that supersedes the maximum temperature required for the preparation of a structurally homogeneous monolith. A novel heat expulsion technique is employed to overcome the heat build-up during the synthesis process. The enormous heat build-up is perceived to encompass the heat associated with initiator decomposition and the heat released from free radical-monomer and monomer-monomer interactions. The heat resulting from the initiator decomposition was expelled along with some gaseous fumes before commencing polymerisation in a gradual addition fashion. Characteristics of a 50 mL monolith synthesized using this technique showed an improved uniformity in the pore structure radially along the length on the monolith. Chromatographic characterization of this adsorbent displayed a persistent binding capacity of 14.5 mg pDNA/mL of the adsorbent. The adsorbent was able to fractionate a clarified bacteria lysate in only 3 min (after loading) into RNA, protein and pDNA respectively. The pDNA fraction obtained was analyzed to be a homogeneous supercoiled pDNA.

Analysing reflective text for learning analytics : an approach using anomaly recontextualisation

Reflective writing is an important learning task to help foster reflective practice, but even when assessed it is rarely analysed or critically reviewed due to its subjective and affective nature. We propose a process for capturing subjective and affective analytics based on the identification and recontextualisation of anomalous features within reflective text. We evaluate 2 human supervised trials of the process, and so demonstrate the potential for an automated Anomaly Recontextualisation process for Learning Analytics.

The Games Concept Approach (GCA) as a mandated practice : views of Singaporean teachers

This paper reports on the views of Singaporean teachers of a mandated curriculum innovation aimed at changing the nature of games pedagogy within the physical education curriculum framework in Singapore. Since its first appearance over 20 years ago, Teaching Games for Understanding (TGfU), as an approach to games pedagogy has gathered support around the world. Through a process of evolution TGfU now has many guises and one of the latest of these is the Games Concept Approach (GCA) a name given to this pedagogical approach in Singapore. As part of a major national curricular reform project the GCA was identified as the preferred method of games teaching and as a result was mandated as required professional practice within physical education teaching. To prepare teachers for the implementation phase, a training program was developed by the National Institute of Education in conjunction with the Ministry of Education and well known experts in the field from the United States. For this part of the study, 22 teachers from across Singapore were interviewed. The data were used to create three fictional narratives, a process described by Sparkes (2002a) and used more recently by Ryan (2005) in the field of literacy. The stories were framed using Foucault’s (1980/1977) notion of governmentality and Bernstein’s (1996) notion of regulative discourse. The narratives reveal tales of confusion, frustration but also of hope and enthusiasm.

A Fixed Order Model Matching Approach for the Design of Robust Power System Oscillation Damper

This book focuses on how evolutionary computing techniques benefit engineering research and development tasks by converting practical problems of growing complexities into simple formulations, thus largely reducing development efforts. This book begins with an overview of the optimization theory and modern evolutionary computing techniques, and goes on to cover specific applications of evolutionary computing to power system optimization and control problems.