Directed phage display

How do the mutations identified by phage display improve binding specificity There is as yet no direct stmctural information on the phage-selected inhibitors however they can be modeled using data from the crystal structures of other Kunitz domains bound to serine proteinases. These studies lead to the conclusion that the mutations identified by phage display improve binding specificity by maximizing complementarity between the... 

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. 

Given this rationale, we have reasoned it would be possible, and have recently reported the direct mapping of the human vasculature by in vivo phage display in a patient [20]. This large-scale survey of motifs that localized to different organs... 

Bach M, et al. Isolation from phage display libraries of lysine-deficient human epidermal growth factor variants for directional conjugation as targeting ligands. Protein Eng 2003 16 1107. 

MAbs directed against the C. neoformans polysaccharide are closely related, and some differ only in a few amino acid residues [142,143]. This fine specificity is related to their protective efficacy. A highly protective IgGl mAh was used to screen phage-displayed peptide libraries, and several binding peptide sequences were identified [144]. The dodecapeptide GLQYTPSWMLVG bound to the mAh with a Kd of 295 nM, measured by surface plasmon resonance (SPR) and inhibited binding of the mAh to the polysaccharide. The dodecapeptide was able to induce only a weak antipolysaccharide antibody response [145]. 

To circumvent some of the limitations of direct immunization, phage display technology has been applied to the preparation of fully human MABs. Gene libraries of cDNA from nonimmune or immunized donor lymphocytes are expressed in bacteriophages. The bacteriophages display functional antibody fragments and can... 

For years, it was thought that the cytosolic carboxyl termini as well as various intracellular loops of the 5-HT receptors bind to FRAPs. Efforts from our lab to identify FRAPs utilized yeast two-hybrid screens see Fig. 1 for an overview), phage display see Fig. 2 for an overview), and direct biochemical approaches (8-13). These studies led to the discovery of many 5-HT2A receptorinteracting proteins, including caveolin-1 (Cav-1), arrestin-2 (Arr-2), arrestin-3 (Arr-3), microtubule-associated protein-1A (MAP-1A), and postsynaptic density protein-95 (PSD-95) see Table 1). 

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