Figure 2. The concept of competitive elution, here used for elution of a monovalent Z-fusion by the competitor ZZ-BB. Source: Modified from Nisson and coworkers (47).

The described strategy has been used to produce the Kle-now fragment of E. coli DNA polymerase I as a Z-Klenow fusion, and the recovered product retained full enzymatic activity (47).

Stabilization to Proteolysis of Labile Gene Products

When expressing mammalian proteins in a heterologous host such as E. coli, proteolytic degradation is one of the more common problems encountered. A review describing the different strategies available to minimize proteolytic degradation of recombinant gene products was recently presented (97). Fusion of a proteolytically labile target protein to stable fusion partners, such as the ZZ and ABP tags, has been shown to have a certain, but limited, stabilizing effect (88,97). However, a much more pronounced stabilizing effect has been obtained when employing a dual affinity fusion concept (Fig. 3) (98), where the protein of interest, X, is fused between the two different affinity tag

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