Protein Purification

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Chapter: Pharmaceutical Engineering: Bioprocessing

The purpose of bioprocess engineering is to utilize resources necessary to pro-mote the growth of microorganisms in a controlled environment for the purpose of producing a product of biological origin.


PROTEIN PURIFICATION

The purpose of bioprocess engineering is to utilize resources necessary to pro-mote the growth of microorganisms in a controlled environment for the purpose of producing a product of biological origin. 

TABLE 16.4 Purification Steps of Soluble Proteins


Note that additional processing is required to purify the product obtained from these systems. The purification from cell culture of soluble proteins may be conducted by combining traditional purification steps (Harrison, 1994). These steps are shown in Table 16.4. Each step focuses on particular physicochemical properties; for example, ion exchange, hydrophobic interaction, and gel chromatography separate molecules based on charge, hydrophobicity, and molecular size, respectively. Inclusion bodies with complex tertiary structures undergo additional steps, including washing, solu-bilizing, and refolding of proteins before further purification steps are adopted.

As with all of the foregoing processes, validation of purification steps is required. Indeed, the sequence of designing protein purification processes may be described as follows (Nelson, 1991):

·       Stepwise recovery yields

·       Impurity removal

·       Scalability of protein purification steps

·       Validation of protein purification steps

Biotechnological innovation occurs as a result of complex integrated bio-processes based on molecular and cellular biology. The key processes have been described briefly in this chapter, and discourses of some length are available on each of these topics in the literature. Since the next generation of pharmaceutical products is likely to be developed by these methods, readers are strongly encouraged to familiarize themselves with these topics and with the more fundamental issues of molecular and cellular biology not covered in this text (Bolsover et al., 1997).

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