Secondary Purification Steps

Depending on the feedstock, the technique selected, and the optimization, the initial capture step can often give purities in the range of 80 to 95%. This is often adequate for in vitro usages. However, for in vivo use, purities in excess of 99% are usually required. In addition to protein contaminants, other impurities such as DNA, endotoxins, viruses, and aggregates need to be removed. In such cases, a multistep procedure is almost inevitable.

All the same techniques used for initial capture can be used for secondary purification. Indeed, many of the techniques, such as HIC and HAP, are used more often as polishing steps than as initial capture steps.

The orientation of these secondary steps tends to be different from the initial capture step. With initial capture, the goal is to remove as many and as much contamination as possible. The secondary purification steps tend to be planned to remove individual contaminants. For instance,

186 antibody purification

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