Antibody phage display technology

An important aspect of antibody phage display technology is the expression of fragments of antibody molecules on the surface of filamentous phage particles. The expressed antibody fragments may take various formats, including Fab fragments, single-chain (sc) Fv fragments and fragments composed of

Figure 1 Phagemid pHENl (A), a phage antibody (B) and a soluble scFv fragment (C). P/O, lacZ promoter/operator; L, pelB leader; VH + Vu, scFv encoding gene; M, Myc tag; A, amber codon; g3, gene3; colE1 ori, E. coli origin of replication: AMP" ampicillin resistance gene; M13 ori, phage intergenic region: g3p, coat protein 3.

fragment. The phagemid harbors an amber stop codon preceding the gene encoding g3p. As a consequence, in SupE suppressor strains of Escherichia coli, the amber codon is read as a glutamine and the antibody is displayed on the phage via a g3p anchor. In nonsuppressor strains, the amber codon is read as a stop codon, and soluble antibody fragments are secreted from the bacteria. Inclusion of the M13 phage intergenic region in the vector allows for replication and packaging of single-stranded phagemid DNA with the aid of helper phage. Because the filamentous particles contain a copy of the pHENl phagemid, selections based on the functional properties of the displayed antibody also result in selection of the genes encoding these antibodies. Phage displayed antibodies may be of any animal species including human, depending on the source of antibody variable (V) regions used for cloning.

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