Phage libraries selection

Barry MA, Dower WJ, Johnston SA. Toward cell-targeting gene therapy vectors selection of cell-binding peptides from random peptide-presenting phage libraries. Nat Med 1996 2(3) 299-305. 

The phage library stock may now be stored at 4°C. However, the selection procedure must be carried out with freshly prepared phage, since the antibody fusion protein may not be stable. To do this, the library can be reamplified by growth on XLl-Blue (Section 3.2.7., steps 7—17). 

Garrard, L. J. and Henner, D. J. (1993) Selection of an anti-IGF-1 Fab from a Fab phage library created by mutagenesis of multiple CDR loops. Gene 128, 103-109. 

The affinity of antibodies selected from naive, synthetic, or even immune phage libraries is typically sufficient for use as a research reagent, but too low for some specific therapeutic applications such as viral neutralization or tumor imaging. For many applications, affinity maturation can be bypassed completely by the construction of multivalent molecules. If this is not sufficient selected antibody clones may be affinity matured (see Figure 18.13). 

Cai X, Garen A, Anti-melanoma antibodies from melanoma patients immunized with genetically modified autologous tumor cells, selection of specific antibodies from single-chain Fv fusion phage libraries, Proc. Natl. Acad. Sci. USA, 92 6537-6541, 19. 

FIGURE 19.6 Biopanning on a column. The target is covalently immobilized on a chromatographic support. The phage library is then applied to the column and, after an extensive washing step, bound phages are recovered by elution, amplified, and submitted to a successive selection cycle. 

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