Phage display libraries, construction

The use of synthetic trinucleotides as building blocks would obviate the problems of stop codons or other unwanted amino acids turning up randomly in hypervariable sequences, and would indeed allow the predetermined definition of amino acid codon frequency bias. This has been demonstrated by Glaser et al. [97] for phage-display library construction see also Ref. 98 for a review of current DNA synthesis technology. 

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,...

A major source of human antibodies are phage display libraries, which are constructed from various genetic sources. Antibodies are expressed as scFV and Fab antibody fragments using various vector systems. This review offers a comprehensive overview of M13 phage display antibody vectors and discusses their applications. 

To complete the library construction, the heteroduplex CCC-dsDNA must be introduced into an E. eoli host that contains an F episome to enable M13 bacteriophage infection and propagation. Phage-displayed library diversities are limited by methods for introducing DNA into E. eoli, with the most efficient method being high-voltage electroporation. 

Sidhu, S. S. and Weiss, G. A. (2004) Constructing phage display libraries by oligonucleotide-directed mutagenesis, in Phage Display A Practical Approach (Lowman, H. B. and Clackson, T., eds.), Oxford University Press, Oxford, UK, pp. 27-41... 

Chemokine inhibitors might need optimization, in order to increase their affinity for a define chemokine or to broaden/restrain their binding profile. In order to optimize the binding profile ofEvasin-4, we constructed a phage display library containing randomly mutated DNA sequences... 

There has been substantial progress in the construction of phage-displayed peptide libraries and in screening methods with the libraries to isolate peptide receptor ligands [66,116-119]. Phage display peptide libraries offer opportunities to characterize peptide binding specificity of important proteins, such as... 

Geoffroy, F., Sodoyer, R., Aujame, L (1994) A new phage display system to construct multicombinatonal libraries of very large antibody repertoires Gene 151, 109-113... 

A large number of different phage display vectors have been constructed. With pretending to be complete, Table 1 lists a selection of phage display vectors. Some of them have not been used for the construction of a library up to now but have been included because they offer possible alternatives. For example, one of the systems allows the success of antibody gene cloning to be monitored by the expression of green fluorescent protein (52). 

---