The concept of phage display is based on insertion of random oligonucleotides at an appropriate location within a structural gene of a bacteriophage. The resulting phage will constitute a library of random peptides displayed on the surface of the bacteriophages, with the encoding genotype packaged within each phage particle. Using a phagemid/helper phage system, the random peptides are interspersed between wild-type coat proteins. Libraries of phage-expressed peptides may be used to search for novel peptide ligands to target proteins. The success of finding a peptide with a desired property in a given library is highly dependent on the diversity and quality of the library. The protocols in this chapter describe the construction of a high-diversity library of phagemid vector encoding fusions of the phage coat protein pVIII with random peptides, from which a phage library displaying random peptides can be prepared.
Fagerlund A, Myrset AH, Kulseth MA (2008) Construction and characterisation of a 9-mer phage display pVIII-library with regulated peptide density. Appl Microbiol Biotechnol 80:925–936PubMedCrossRefGoogle Scholar
Noren KA, Noren CJ (2001) Construction of high-complexity combinatorial phage display peptide libraries. Methods 23:169–178PubMedCrossRefGoogle Scholar
Göbel U, Maas R, Clad A (1987) Quantitative electroelution of oligonucleotides and large DNA fragments from gels and purification by electrodialysis. J Biochem Biophys Methods 14:245–260PubMedCrossRefGoogle Scholar
Gründemann D, Schömig E (1996) Protection of DNA during preparative agarose gel electrophoresis against damage induced by ultraviolet light. Biotechniques 21:898–903PubMedGoogle Scholar
Bonnycastle LL, Mehroke JS, Rashed M, Gong X, Scott JK (1996) Probing the basis of antibody reactivity with a panel of constrained peptide libraries displayed by filamentous phage. J Mol Biol 258:747–762PubMedCrossRefGoogle Scholar