Bacteriophage Display of Combinatorial Antibody Libraries


Bacteriophage display of combinatorial antibody libraries is one means by which monoclonal antibodies of a desired specificity can be selected without the use of conventional hybridoma technology. Phage display is the expression of proteins and peptides on the surface of a bacteriophage particle, and a combinatorial antibody library is a collection of rearranged variable‐region gene segments that have been randomly combined through the use of recombinant molecular biology techniques to yield a large ‘library’ of antibodies.

Keywords: phage display; antibody library; fab; scFv

Figure 1.

Expression of antibody fragments by phage display. A schematic representation of the expression of Fab and scFv antibody fragments using phage display. The antibody fragments are cloned into the phagemid vector under the control of the lacZ promoter as fusion proteins with the C‐terminus of the minor coat protein, gene III. The translated proteins are directed to the periplasmic space by the pelB signal peptide where they are assembled on the phage particles. (a) Fab display. (b) scFv display.

Figure 2.

Immunoglobulin structure. A schematic representation of the structure of an antibody molecule. The intact IgG molecule contains two identical heavy chains and two identical light chains that are covalently linked through disulfide bonds. The Fab molecule consists of a single light chain and the heavy chain Fd (VH+CH1) covalently linked by a single disulfide bond. The scFv fragment consists of a light chain variable region linked to a heavy chain variable region through a synthetic peptide linker.



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Further Reading

Burton DR and Barbas CF III (1994) Human antibodies from combinatorial libraries. Advances in Immunology 57: 191–280.

Huse WD, Sastry L, Iverson SA et al. (1989) Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science 246: 1275–1281.

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Persson MAA, Caothien RH and Burton DR (1991) Generation of diverse high‐affinity human monoclonal antibodies by repertoire cloning. Proceedings of the National Academy of Sciences of the USA 88: 2432–2436.

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Rader C and Barbas III CF (1997) Phage display of combinatorial antibody libraries. Current Opinion in Biotechnology 8: 503–508.

Söderlind E, Simonsson AC and Borrebaeck CAK (1992) Phage display technology in antibody engineering: design of phagemid vectors and in vitro maturation systems. Immunological Reviews 130: 109–123.

Winter G and Milstein C (1991) Man‐made antibodies. Nature 349: 293–299.

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Andris‐Widhopf, Jennifer, Steinberger, Peter, and Barbas, Carlos F(Apr 2001) Bacteriophage Display of Combinatorial Antibody Libraries. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0001119]