Unique Purification Strategies

Sztajer and Bryjak [48] have taken an entirely different approach to the purification of the lipase from Pseudomonas fluorescens by investigating the use of ultrafiltration capillary membranes. A two-step procedure involving continuous fractionation of the protein on polyacrylonitrile membrane followed by concentration on polysulfone membranes is suggested for continuous lipase recovery. They observe that the permeate fluxes through both membranes are similar, thereby suggesting that changing the production scales should not be difficult. 

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This chapter begins with an introduction of general considerations for protein purification and then focuses on purification strategies and characterization technique for MCOs. Examples of MCO purification and characterization will regularly be referenced and the chapter will pay particular attention to the characterization of metals centers, as expression, purification, and characterization of copper-dependent MCOs require some unique methods. Techniques to characterize MCO activity and behavior after immobilization onto electrode surfaces will not be covered, as these techniques are described elsewhere in the book. 

After the extraction of the target biomolecule, purification is the next step. It is possible to select the entire sequence of purification steps for isolating a particular protein based on its properties, and on the fact that every protein presents a unique combination of characteristics. A strategy should be established to minimize the number of stages, leading to the maximum protein yield, minimum cost, and the required purity for the product s final application. 

Depending on the application of the antibody, purity and recovery are likely to determine the strategy of purification. In this chapter, common chromatographic methods for antibody purification will be described. In general, multiple-step purification and combination of methods are required to obtain sufficient purity of antibodies. Because every antibody is unique and each has a distinctive distribution of hydrophobic and positive and negative charge characteristics, modification and optimization of purification procedures might be necessary [2]. 

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