Libraries phage display

A further advance in antibody technology is the development of transgenic mouse human strains. XenoMouse animals have been engineered in such a way that they now produce exclusively human antibodies rather than murine antibodies when immunized. The use of XenoMouse animals to produce MAbs avoids the need for any engineering of the antibody genes, since the products are already 100% human protein. XenoMouse animals are fully compatible with standard hybridoma technology and can be readily adopted by laboratories experienced in monoclonal antibody production [56]. 

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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,...

Palzkill, T., Huang, W., and Weinstock, G. M. (1998). Mapping protein-ligand interactions using whole genome phage display libraries. Gene 221, 79-83. 

Vaughan, T. J., Williams, A. J., Pritchard, K., Osbourn, J. K., Pope, A. R., Eamshaw, J. C., McCafferty, J., Hodits, R. A., Wilton, J., and Johnson, K. S. (1996). Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library. Nat. Biotechnol. 14, 309-314. 

Pasqualini R, Arap W, Rajotte D et al. In vivo selection of phage display libraries. In Phage Display A Laboratory Manual (Barbas CF III, Burton DR, Scott JK, Silverman GJ, Eds.). New York Cold Spring Harbor Laboratory Press 2000, 22.1-22.24. 

The diversity of antibodies can be increased further with phage display libraries and recombinant DNA technology. In recombinant DNA... 

Willats WGT, Gilmartin PM, Mikkelsen JD, Knox JP. Cell wall antibodies without immunization generation and use of de-esterified homogalacturonan block-specific antibodies from a naive phage display library. Plant J. 1999 18 57-65. 

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. 

Koivunen E, Gay DA, Ruoslahti E. Selection of peptides binding to the alpha 5 beta 1 integrin from phage display library. J Biol Chem 1993 268(27) 20205-20210. 

De Martino, T., Minenkova, O., Bombardi, V, Anastasi, A. M., Lindstedt, R., Felici, F., De Santis, R., Verdoliva, A. Identification and refinement of a peptide affinity ligand with unique specificity for a monoclonal anti-tenascin-C antibody by screening of a phage display library. / Chromatogr A 2006, 1107, 182-191. 

A number of molecules in groups 2 and 3 have been identified by the differential homing capacity of phage display libraries and combination peptide libraries [71]. Biochemical strategies such as the application of 2D gel electrophoresis on protein extracts from endothelial cell surfaces have also proven useful in this respect [72]. 

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