Preliminary separation steps

At this stage, the method of extraction must be selected. Unless dealing with a secreted protein in cell culture, any extraction from a biological source will require mechanical disruption of cells. This leads to dispersion and dilution of cell contents resulting in release of proteolytic enzymes and general acidification. Appropriate buffer solutions can be used to guard against rapid acidification and addition of sucrose or maltose may help to stabilize lysosomal membranes and hence reduce the release of proteases.

It is advisable to maintain the ionic strength of the medium to aid protein stability. Many other additives can be included at this stage, e.g. protease inhibitors, or EDTA to chelate metal ions which can catalyse oxidation. However, with prior knowledge of the protein stability these can be kept to a minimum as they will inevitably have to be removed at a later stage and may cause unnecessary complications and delay. As a general rule, the preliminary extraction should be performed quickly, at sub-ambient temperatures.

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