Comparison with recombinant proteins

In the present context, fluorescence is an important tool for the rapid comparison of a protein with a standard sample of the recombinant or the authentic protein, as part of the process of characterizing and authenticating a recombinant protein. Coupled with measurements of the depolarization of fluorescence, the method can provide information on the dynamics of a protein and on the change in dynamics in response to, for example, binding of ligands. 

Figure 8.26 illustrates the application of the first method to an antibody directed against the recombinant protein bFGF. Twelve peptides are identified in the MALDI spectrum of the mixture obtained after enzymatic cleavage non-bounded (Figure 8.26A). After immunoprecipitation with the antibody, the peptides are washed out, and only four peptides are detected as shown in Figure 8.26B. A comparison of the sequences of those peptides allows the epitope to be located somewhere in the residues 15-27, as explained in Figure 8.26C.

Mande SC, Mainfiroid V, Kalk KH, Goraj K, Martial JA, Hoi WG. Crystal structure of recombinant human triosephosphate isomerase at 2.8 A resolution. Triosephosphate isomerase-related human genetic disorders and comparison with the trypanosoma enzyme. Protein Sci 1994 3 810-21. 

For a recombinant DNA protein, expression system comparison with the native sequence. 

Once the expression cassette has been optimized, highly diluted agrobacterial suspension carrying the cDNA of the TMV-based vector is used for large-scale agroinfiltration of whole tobacco plants. The biomass can be harvested 7-10 days later and used for isolation of the recombinant protein of interest. In comparison with other systems, this protocol is extremely fast, provides high yields, and has the potential for unlimited scalability. In addition, our... 

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