ABSTRACT
The development of methods and techniques for the separation and purification of biological macromolecules such as proteins has been a prerequisite for advances in bioscience and biotechnology during the last five decades. The recovery and purification of a bioproduct is carried out to isolate it
from its production system (cell culture, plant or animal tissues), and to obtain the required purity and formulation. Before establishing a strategy for recovery and purification, it is essential to collect all available information related to the protein and to the medium where it is found. Usually, not only the theoretical information, but also preliminary experiments are needed. It is also noted that the feasibility of a process on a laboratory scale does not guarantee its feasibility on an industrial scale. With the use of high performance materials and automated instruments,
protein separation is becoming a more controllable process. However, some problems persist even with the use of sophisticated instruments. Many difficulties are still found in determining the optimal extraction and purification conditions, as well as in selecting suitable methods for detecting the protein and quantifying its biological activity. A general scheme of the recovery and purification process for a
bioproduct is shown in Figure 12.1. Despite the lack of a universal strategy, some general directions (listed below) should be followed when establishing the purification protocol:
(i) choose separation methods based on different physical principles; (ii) choose methods based on physical properties related to the greatest
differences between products and impurities; (iii) first remove contaminants that are most dissimilar to the product
and/or those that are most abundant; (iv) assign the most demanding process to the end of the sequence.
