In this work, we designed and built a continuous crystallisation oscillatory flow platform. The lysozyme crystallisation behaviours were investigated at concentrations from 30 to 100 mg/mL, under oscillatory conditions with amplitude (0) from 10 to 25 mm and frequency () from 0.05 to 0.25 Hz in a batch oscillatory flow crystallisation platform. The nucleation rate increased with increase in concentration of initial lysozyme solution, and was also found to increase with increase in shear rate. By learning the thermodynamics and kinetics of lysozyme crystallisation in batch oscillatory flow, the batch crystallisation process was successfully transferred to a continuous oscillatory flow crystallisation process. The equilibrium state of continuous crystallisation reached at residence time 200 min, and the final product crystals shape and size were consistent during the continuous process. This work demonstrates the feasibility of oscillatory flow based platforms for the development of continuous protein crystallisation as for downstream bioseparation.
The objective of the presented module is to train students with no background in process development and scale-up of chromatographic processes to a high level of competency within forty contact hours. The key pedagogical approach is 'progression' where students' capabilities are gradually built up with appropriate scaffolding provided at each stage of their learning. The module is broken up into three steps with each step covering a different aspect of chromatography. Knowledge gained in one step is the foundation for work in the next. In the first step students investigate several chromatographic column packing materials and perform a solvent selection process. Design of Experiment (DOE) to systematically vary process parameters for method development is introduced in the second step. In the last step, students use a preparative-LC system to perform a larger scale separation. Students explore different scale-up scenarios, including volume fraction collection and column overloading. Pedagogic outcomes of the module were determined through surveys, interviews and personal interaction during the study. Results clearly indicate that students engaged well with the module while meeting overall learning objectives. The module is equally suitable for third or fourth year university students or industry practitioners unfamiliar with chromatography as part of continuing professional development.
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