Phage antibody libraries

Sblattero, D., and Bradbury, A. (2000). Exploiting recombination in single bacteria to make large phage antibody libraries. Nat. Biotechnol. 18, 75-80. 

De Kruif, J., Terstappen, L, Boel, E, and Logtenberg, T (1995) Rapid selection of cell subpopulation-specific human monoclonal antibodies from a synthetic phage antibody library Proc Natl Acad Sci. USA 92, 3938—3942... 

Sheets, M. D., Amersdorfer, P., Finnem, R., Sargent, P., Lindqvist, E., Schier, R., et al. (1998) Efficient construction of a large nonimmune phage antibody library the production of high-affinity human single-chain antibodies to protein antigens. Proc. Natl. Acad. Sci. USA 95, 6157-6162. 

Making and Selecting from Phage Antibody Libraries... 

Although a number of different phage (mid) antibody libraries have been published (see Table 18.1), the number which are naive is relatively small. To date, all phage antibody libraries have been made on the filamentous phage, Ff (which comprises fd, fl, and Ml3). In the creation of such libraries a number of different choices can be made ... 

The choice of which form of antibody fragment to use in a phage antibody library (Figure 18.5) is on the whole dictated by practical considerations (Table 18.4). 

Many different linkers have been used in single antibodies expressed as scFv (Table 18.7). However, although some comparisons have been made for single scFvs, no direct comparison has been made between these different linkers in the creation of phage antibody libraries. In fact, the only linker which has been used extensively in scFv libraries is the (Gly4-Ser)3 linker. This has a number of... 

This discussion has focused on the desire to create a one-pot naive phage antibody library. However, phage display has also been used very effectively to isolate binding antibodies when libraries are made from immunized experimental animals [32], or from naturally immunized humans [7, 30] in these cases the V-gene source being either spleen or peripheral-blood lymphocytes. 

V genes to make single-pot phage antibody libraries. 

The isolation of an antibody is an extremely useful first step toward understanding the function of the protein to which it binds. scFvs derived from phage antibody libraries have been used in immunofluorescence, immunoprecipitation, fluorescence-activated cell sorting, Western blotting, and inhibition of function studies, both in vivo, in tissue culture cells, and in vitro. In this sense, they can essentially be used in the same way as conventional hybridoma-derived antibodies. They have the advantage, however, that the genes for the variable regions are cloned simultaneously with selection. This allows the fusion of functional elements, such as dimerization domains, effector or detector functions to selected scFvs [38], the re-creation of complete... 

Chapter 18 Making and Selecting from Phage Antibody Libraries Andrew Bradbury, Daniele Sblattero,... 

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