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Review
. 2004 Sep;57(9):912-7.
doi: 10.1136/jcp.2003.014407.

Demystified...recombinant antibodies

K A Smith  1 , P N Nelson, P Warren, S J Astley, P G Murray, J Greenman
Affiliations
Review

Demystified...recombinant antibodies

K A Smith et al. J Clin Pathol. 2004 Sep.

Abstract

Recombinant antibodies are important tools for biomedical research and are increasingly being used as clinical diagnostic/therapeutic reagents. In this article, a background to humanized antibodies is given, together with details of the generation of antibody fragments--for example, single chain Fv fragments. Phage antibody fragments are fast becoming popular and can be generated by simple established methods of affinity enrichment from libraries derived from immune cells. Phage display methodology can also be used for the affinity enrichment of existing antibody fragments to provide a reagent with a higher affinity. Here, phage antibodies are demystified to provide a greater understanding of the potential of these reagents and to engage clinicians and biomedical scientists alike to think about potential applications in pathology and clinical settings.

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Figures

Figure 1
Figure 1
(A) Structure of a human antibody, (B) Fab fragment, and (C) a single chain Fv fragment—scFv. VH, variable region of heavy chain; VL, variable region of light chain; CH, constant region of heavy chain; CL, constant region of light chain.
Figure 2
Figure 2
The structure of a phage particle (left) and the structure of a phage particle where a single chain heavy and light chain variable region fragment (scFv) is expressed as a fusion to the pIII coat protein (right).
Figure 3
Figure 3
Selecting antibodies from phage libraries. (1) Phage are incubated with the antigen of interest—any phage that express a single chain heavy and light chain variable region fragment (scFv) that binds to the antigen. (2) Phage that express scFv fragments that do not bind to the antigen are washed away. (3) Phage that express scFv fragments that bind to the antigen are isolated, expanded, purified, and then used in further applications.
Figure 4
Figure 4
Immunohistochemical staining in a colon carcinoma using a phage displayed antibody specific to vascular endothelial growth factor165. Original magnification, ×40.
See this image and copyright information in PMC

References

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