This chapter provides a more detailed overview of the equipment requirements for protein purification and the practical aspects of developing a purification strategy, including the ordering of unit operations, buffer preparation, and approaches to minimizing yield losses. A well-equipped laboratory is an essential prerequisite to successful protein purification and time spent in deciding needs and purchasing essential items will help to avoid panics mid-way through a protocol when a key piece of equipment is lacking. This said, one should avoid expensive purchases (unless budget is no object) as the majority of purifications can be achieved with fairly routine equipment. The few essential pieces of expensive equipment are a spectrophotometer, a centrifuge, and a chromatography set-up. In general, money is best spent in purchasing plenty of the cheaper items such as tubes, beakers, measuring cylinders, salts, and buffers. Chromatography equipment is an essential item for any purification laboratory. There are plenty of expensive chromatography set-ups available which can provide a remarkable saving in time for process development purposes. If you are not familiar with such equipment and are new to the world of protein purification, I suggest you make do with a simple chromatography set-up to start with, until you become familiar with the technique and learn more about your exact requirements and how alternative tailor-made process development kits differ. A protein purification laboratory should be equipped with supplies of tapwater, de-mineralized water, and distilled water. Electricity and sinks are taken for granted. Required equipment can be roughly grouped into three categories: • those for ancillary purposes • those for detection • those for separation Table 1 lists the essential items for the protein purification laboratory. Time and money spent wisely on buying adequate supplies of support materials will pay dividends. There is nothing more irritating than having to rush around in search of a clock or some salt half-way through a delicate purification with your enzyme degrading in front of your eyes! Key requirements are tubes, beakers, pipettes, stirrers, and timers. In addition essential chemicals include salts and buffers. Adjustable pipettes (e.g. Gilson) are essential and those suitable for sample volumes from 10 μl to 5000 μl are recommended.
Optimization of Reaction Selectivity Using CFD-Based Compartmental Modeling and Surrogate-Based Optimization
