Proteins phage display selections

Chang, W. J., Kishikawa-Kiyama, M., Shibata, Y., Lee S. Y., Abiko, Y. (2004). Inhibition of Porphyromonas gingivalis hemagglutinating activity by synthetic peptides derived from phage display selection using MAb against the recombinant outer membrane protein. Hybrid Hybridom., 23, 352-356. 

Keresztessy, Z., Csosz, E., Harsfalvi, I., et al. (2006). Phage display selection of efficient glutamine-donor substrate peptides for transglutaminase 2. Protein ScL, 15, 2466-2480. 

Phage Display Selections against Purified Proteins, 95... 

Similarly small protein ligands have been created by selection from randomized Protein A receptor domain nsing phage display selection techniques from about a 40-million-member library (69). The selected peptides, termed affibodies, were so stable that the peptides, immobilized on HiTrap NHS Sepharose, could be repetitively sanitized by treatment with 0.5 M sodium hydroxide. 

Figure 17.10 Construction of a two helix truncated Z domain, (a) Diagram of the three-helix bundle Z domain of protein A (blue) bound to the Fc fragment of IgG (green). The third helix stabilizes the two Fc-binding helices, (b) Three phage-display libraries of the truncated Z-domaln peptide were selected for binding to the Fc. First, four residues at the former helix 3 interface ("exoface") were sorted the consensus sequence from this library was used as the template for an "intrafece" library, in which residues between helices 1 and 2 were randomized. The most active sequence from this library was used as a template for five libraries in which residues on the Fc-binding face ("interface") were randomized. Colored residues were randomized blue residues were conserved as the wild-type amino acid while yellow residues reached a nonwild-type consensus, [(b) Adapted from A.C. Braisted and J.A. Wells,...

EMPl, selected by phage display from random peptide libraries, demonstrates that a dimer of a 20-residue peptide can mimic the function of a monomeric 166-residue protein. In contrast to the minimized Z domain, this selected peptide shares neither the sequence nor the structure of the natural hormone. Thus, there can be a number of ways to solve a molecular recognition problem, and combinatorial methods such as phage display allow us to sort through a multitude of structural scaffolds to discover novel solutions. 

Protein engineering is now routinely used to modify protein molecules either via site-directed mutagenesis or by combinatorial methods. Factors that are Important for the stability of proteins have been studied, such as stabilization of a helices and reducing the number of conformations in the unfolded state. Combinatorial methods produce a large number of random mutants from which those with the desired properties are selected in vitro using phage display. Specific enzyme inhibitors, increased enzymatic activity and agonists of receptor molecules are examples of successful use of this method. 

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